S UMMA RY OF WORK

' Drugs serve as positive reinforcers to maintain and str ength en behavior leading to thei r ad m inistration and a I n s . m can control beh vior through their ability to function as discriminative sti muli many situations , d g of abuse function throug h pha rmacological and behavioral mechanisms to persistently sustai n long seq uences k d m - of d ru g see ing behavior that are very resistant to extinction . These long seq uences of g seeking behavior can be analyzed using schedu le-controlled performances i n the same way as opera nt behavior v k U - maintained by oth er e ents such as food or shoc . sing a variety of intravenous self adm inistration i v proced ures in rats and pr mates , ongoing experiments are being conducted to e aluate behavi or maintained by d rugs and the a bility of pharmacological treatments , antagonist adm in istration or the development of dependence) and/or behavio ral manipulations to modify d rug self-administration behavior and/or food

- . will n rv mai ntained behavior These stud ies compare respond ing maintained u der fixed rati o , fixed i nte al and l - i comp ex second order schedules , by var ous drugs includ ing cocaine , nicotin e and other psychomotor seda tive/a nxiol tics -9 stimu lants , benzod iazepi nes and other y , morphine and other opioids and delta THC (the

t . F ac ive i ngredient of marijuana) or example , since serotonergic mechanisms appear to u nd erli e l psychomotor sti mu ant action , recent stud ies evaluated the effects of sertral ine , a selective serotonergic k f i upta e in hi bitor that is effective as a n a ntidepressant , on th e reinforcing ef ects of v . nicoti ne in sq u i rrel

. I n v monkeys addition to differences in the pharmacological efficacy of d rugs to control or modify beha ior, it is clea r that behavioral and environmental factors play an important role i n the contro l that even h ig h ly

- efficacious d r ugs exert on behavior. The focus of exper i ments in the rhesus self admi n istration la b are to study the pharm acolog ical , behaviora l , and environmental varia bles involved in initiating and maintaini ng

- - dmg self adm inistrati on . Recent studies assessed rates of acq uisition of cocaine self i njection in rh esus mon keys found a relationship with behavioral history , but not activity level . Additional studies a re eval uating the effects of serotonerg ic antagonists on delta-amphetam ine self- injection and noradrenerg ic antagonists “ rs - I . n w and uptake i n hi bito on cocai ne self injection another study , e are evaluating the reinforci ng efficacy of I- t I - the stereoisomers of deprenyl (selegil in e) in comparison to me hamphetamine and its stereoisomer, as well - as eval uating the a bility of deprenyl pretreatment to alter self adm inistration of coca ine , metham phetam ine or beta -phenylethylamine

v PHS 6040 (R e . PU BLI CATIONS

- S Y S . B v m l Gold ber g R , asar eha ioral phar acology of selegil in e ( deprenyl) and its metabolites and i nt eractions - B B . with ph enylethylam i ne , P rogress in rain Research ; in press

C linica l - S Youd im M B . l Gold berg R , H and preclin ical experience with deprenyl (selegi lin e) with regard to a buse P 1 994 l iability , C li nical ha rm acology and Therapeutics ; in press .

- - W Y S N SS S . I v i l inger G , asa r , eg us , Goldberg R ntra enous self admin istration studies w th deprenyl (selegiline) k P 1 994 in mon eys , Cl in ical harmacology and Th erapeutics ; i n press .

- - Y S Y B J . A k I r asar , ergman mphetami ne li e effect of deprenyl in d ug discrim ination studies , Cl i nical P c 1 994 ha rm acology and Therapeuti s ; in press .

S u A A NA G ri . B m a e ca m il I S b . anner d C , tor , ffiths RR ehavioral phar acology of i n ba boons n : tephens DN ed . A l B - a b : Ps cho h a rrn a col o S ri B S nxio ytic c r olines from molecular biology to the clinic . y p gy e es I I . erl in : pri nger - l 20 . Ver ag ,

M G K E S M K BJ P KL S u A S ilver ma n K G umford , vans , , am inski , reston , anner d C , , , riffiths RR . Discr i m i native

t f Ps ch o h a rrn a colo - stimu lus a nd su bjec ive effects of theobrom ine and caf eine i n hu mans , y p gy 8 .

B J i - S A K k G . M H 1 annerud C , a m ins i , r ffiths RR aintenance of antagon ists self i njection i n baboons , Ex erimenta l Ps cho h a r rn a col o 1 99 p and Cli nical y p gy 4; in press .

B J i - S A K G . S annerud C , ami nski , r ffiths RR elf injection of phenethyl a mine designer d ru gs i n ba boons B P m 1 994 ehavioral har acology ; in press . SUMMARY O F WORK

Commercia l assays fort he detection of drugs of a buse in urine change periodically and must be reevaluated for perform ance . Stud ies are desig ned to test the validity of new assays with clin ica l specim ens obtai ned from d rug users under controlled conditions .

I Healthy male volunteers with a history of chemica l su bstance a buse participate in these studies . nform ed I B consent is obtained and all procedures are approved by the hospital nstitutional Review oard . Comm ercial assays for detection of d ru gs of a buse in urine are tested for validity with speci mens col lected und er A controlled dosing conditions var iety of drugs of a buse were studied at various dose l evels . Th e resu lts of toGC/ the assays were com pared MS analysis .

These studies test th e validity of commercial assays on clinical samples instead of spiked sam ples a nd provide uniq ue and valua ble information to the mil itary and industry concern ing their ti me course of detection , specificity and accuracy . PU BLI CAT I O NS

M u O . k L J . F i U S . P M F E . J . Cone , , Dic erson , aul , and itchell , orens c Dr g Testing or piates V rine Testing - F r om M i A W O . . A . . 1 7 : 1 56 1 64 or H e , orph ne nd Codein e ith Commercial piate l mm unoassays J nal Toxicol , ,

1 993 .

- J k M . H A . E . . O S J L . C . rw W D . EZ A . M : en ins , ills , , Da in , uestis , and Con e J Validity Testing f Th e creen 1 7 : - 292 298 1 993 . J . . Canna binoid Test . Anal . Toxicol , ,

r S . L . W R . Fr a n ke nfie ld E . J . S . . I Joseph , T , Dicke son , , ill is , , , D Cone , and m ith D R nterference by - N i A i infla mm a tor Dm s M I T A A . . A . I n P onstero dal nt y g in E T and Dx ssays for Drugs of buse J na l Toxicol . ress

1 994 .

D . t . . J . M . c J rw W . M A M J . E . J . a cu/PI N CH enkins , A . , Da in , , H ues is , , Cone , and itchell , Valid ity Testing of the 1 994 T J A . . I P THC est . . nal Toxicol , n ress , . SUMMARY OF WORK

The behavioral pha r macologica l profil e of a drug in a pertinent species is necessary to evaluate quantitatively how the d rug fu nctions as a reinforcer or a pun isher as well as to establish its sti m ul us effects . Sched ules of food presentation with both fixed - interval and fixed- ratio com ponents have been used most frequently i n th is type of study since they generate a wide range of rates and patterns of respond ing withi n a - a ls . session a nd provide sta ble , long term baselines for chron ic studies in individ ual a nim Th e present project involves the assessm ent of both the acute and chronic effects of a variety of drugs on schedu le-controll ed behavior . We have recently shown that the enhanced sensitivity observed to the behaviora l effects of the GA BA er ic opioid antagonist naltrexone is influenced by g processes , and in pa rticular by an action at the

ABA . F rt G associated chl oride channel u her , this sensitivity may be associated with cha nges i n both m u and delta opioid recepto r s . Genetic factors are known to influence the behavioral effects of a nu m ber of different u n dr gs , and studies of the i nteractions betwee environmental and genetic components th at potentially affect l the deve opment of behaviora l tolerance and sensitivity are being initiated . B ehaviorally active drugs can l ri - a so serve as d isc m inative sti mu li to guide behavioral choice . Two and three lever d ru g discri mi nation projects in the la boratory have helped to define and characterize the spectrum of behavioral effects

r - prod uced by the d ug , to compare a range of other compounds , such as cocaine , l deprenyl , morphi ne ,

- midazolam , and caffei ne to characterize the relative potency and efficacy to produce d rug li ke effects , and to ' l ev S - eval uate the d rug s m echan isms of action at the receptor el . i nce most human drug taking behavior

- - involves ch ronic long term use of an ill icit drug or non medical a buse of a prescribed medication , th e conseq uences of ch ron ic admin istration of drugs on schedu le-control led behavior and th e d iscrim inative functions of d rugs are being evaluated . Although the development of tolerance and depend ence are related l F exa m le ' th e to pharmacologica factors , tolerance can also be modified by environmental factors . or p . interaction between drug adm inistration and the ability to perform the task can result i n d ifferential tolerance t that is a fu nc ion of ch ronic daily dose and duration of treatm ent . PU BLI CATIONS

L A JG . S A M R S e rd ikoff S A . S anneru d C , a rl ey J , , lastra Cohen C , Gold berg , R Tolerance to the behavioral t J effects of ch lordiazepoxide : pha rmacological and biochemical selec ivity , ou rn al of P h armacology and

- Exper i mental Therapeuti cs 20.

v S r A Y AM . L k Gau in DV , anne ud C , oung ac of acute tolerance to the discri minative sti m u l us effects of 1 994 L S . mo r phin e i n pigeons , ife ciences ; in press

S A A NA i - 1: annerud C , tor . Drug discr mination analysis of midazolam under a three lever procedure dose n P m dependent differences i n generalization and antagonism , Jour al of har acology a nd Experim ental

Therapeutics 1 994; in press .

' s ri P AM S W O H a r a B F B k M U G . B : ers ico , chi ndler C , , rannoc T , hl R rain tran c ption factor expression effects of - M B 1 00. acute and chron ic am pheta mine and injection stress , olecu lar rain Research

S W P r AM U G G S B - chindler C , e sico , hl R , oldberg R . ehavioral assessment of h ig h dose a mph etam in e P m B r withdrawal : impo rt ance of train ing and testing conditions , har acology , iochemist y and Behavior 1 994; in press .

Y S W B G S - i asar S Y , ch indler C , Thorndi ke E , oldberg R . Eval uation of deprenyl for coca i ne li ke d iscr m inative E J P - 50 stim ulus effects in rats , uropean ou rnal of ha rmacology 2 .

R J T h omd ike - G w M M n E G S S CW. GA A i e iss V , a ey , B , old berg R , chindler B receptor l in ked ch lor d e chan nels f P B 1 994 and the behavioral ef ects of naltrexone in rats , harm acology . iochemistry and B ehavior ; in press .

P AM S l Z M U G B i W B . ersico , ch ind er C , aczek R , ran nock T , hl R rai n transcr ption factor gene expression ,

' neurotransmitter levels and novelty response behaviors: alterations d un ng rat a m ph eta m i ne withdrawal a nd l S 1 994 fol owing chron ic injection stress , ynapse ; i n press .

- B F DM L I B MT DD M offa tt A KI a unber BA S W O Hara , Donovan , indberg , rannock , Ricker f C , g , ch indler C , J R . Pr oenke h a lin Chang TS , N elson , U hl G R p transgenic mice : a sho rt promotor confers h ig h testis expression M - and red uced fertility , olecu lar Reproduction and Development 84 . SUMMARY OF WOR K

Card iovascu la r effects of cocaine are being extensively studied using various species of a nima ls as m odel we l l systems . Recent investigations have focused on the coca ine meta bol ites and pyrolysis products , as as

dm . W g interactions ith repeated use of coca ine , the effects of th e metabolites may play a more promi nent effect " of . W k role in the overal l cocaine ith the widespread use of crac cocaine , the possi bility that the ' pyrolysis products may contri bute to cocaine s cardiovascular effects m ust also be considered . I n studies with anesthetized ra bbit preparations , i njections of smal l doses of cocaine in the hind bra i n of th e ra bbit a produced clear decreases in heart r te , blood pressure and respiratory rate . The sam e d oses g ive i . v . were

- without effect , indicating that these depressant effects of cocaine were mediated in th e h i nd bra i n . Th e coca eth le ne cocaine meta bol ite y had similar effects , althoug h its effect on respi ration was of longer “

. N duration orcocaine d ecreased blood pressure but did not affect respiration . The cocaine pyrolysis l rt products also d ecreased b ood pressure and hea rate but increased respiration . The effects of these e iv compounds were comparable wh en delivered either into the hind brai n or wh n given , i ndicati ng that their effects were not mediated in the hi nd brain . These results suggest that the cocaine pyrolysis products do not w share a com m on mechanism of action with cocaine . Coca ine is often used in com bi nation ith other d rugs . O - ne freq uent drug use combi nation is that of heroin and cocaine . When given to anesthetized ra bbits as a

sl ow iv . , contin uous infusion , cocaine decreased hea rt rate a nd blood pressure a nd increased respiratory a l rate at higher doses . Heroin so produced decreases in blood pressu re and hea rt rate wh ile decreasi ng

. w respiration When g iven i n com bination , the hemodynamic effects ere exacerbated whil e cocai ne fai led to antagon ize th e respiratory depressant effect of heroin . These resu lts suggest that this d rug com bination may t lead to greater adverse effects tha n would be predic ed from the effects of these d rug a l one . PUBLI CATIONS

K A A N A H S . S W . . Erzou ki H , llen C , ewman , Gold berg R chindler C Effects of cocaine coca ine metabolites and n v cocain e pyrolysis products on th e h ind brain cardiorespiratory centers of the ra bbit , Jour al of Cardio ascular

Pharmacology 1 994; in press .

B l S S W . Erzouki H K , aum , G old berg R , ch indler C Comparison of the effects of cocain e and its metabol ites on

- J Ca r d iova s P r 63 . card iovascular function in anesthetized rats , ha macol

S S W S . : Tella R . ch indler C , Goldberg R Cocaine Cardiovascular effects in relation to th e inh i bition of k P m peripheral neurona l monoam ine upta e and central stimulation of the sympathoadrenal system , J har acol

- Exp Ther 62 .

S W W G A Foltin R W F MW N L G r S ch indler C , ilkerson RD , i llis R , , ischman , ewli n D , evin H R , old be g R . l : I n: m L . . P u Card iovascular effects of cocaine u nder ying mechanisms . Ha s ed roblems of dr g dependence N A 1 40 1 994 N I I . I M P 94 1 993 , Volume ationa l nstitute on Drug buse Research onograph , ; H u bl ication no . - 83 .

S T S P J G r S s ' chindl er CW , ella R , rada , oldbe g R . Calciu m channel blocker antagon ize som e of coca i ne s ' l a lter J P m l E T 1 994 ca rdiovascu ar effects , but fail to cocaine s behavioral effects , har aco xp h er ; i n press .

S S Erzou ki H K G S P m ' v chindler CW , Tel la R , , old berg R . har acological m echan isms in coca ine s ca r d io ascu lar A 1 994 effects , Drug and lcohol Dependence ; in press .

S J M T l l S N G G N w A benza ze ine JH W . n hah , itki n , e a R , owak , eorge C , e man H Q uater ary p a nalogs lack - D i rs M - affinity for dopam ine recepto , ed Chem Res 1 994; 88 .

- . J H C S W G S . I : J J K chi ndler, C , old berg R Conditioned withdrawal n affe , Clayton RR , ohanson E, u ha r MJ ,

M MH S l M . u 9 4 . oore , e lers E , eds Th e encyclopedia of dr gs and alcohol , 1 9 ; in press

T S G b S . M Ps chO ha rma colo ella R , old erg R onoam in e uptake i nhi bitors alter pharmacokinetics of coca in e , y p gy - 502 . SUMMARY OF WORK

The presence of dru gs of a buse in saliva of human subjects after dru g adm inistration was stud ied to m w l deter ine th e feasi bil ity of d rug testing ith saliva as the bio ogica l specimen . Healthy su bjects with a I a l l history of ch em ical su bstance a buse vol unteered for these studies . nform ed consent was obta ined a nd d v I B F proce ures were appro ed by the hospital nstitutional Review oard . ollowing the ad mi nistration of ri B v cocai ne , ma juana or opiates , saliva and blood samples were collected periodically . eha ioral and S physiological measures were made concurrently with collection of biological fluids . am ples were analyzed ch r oma to / S ' by radioimm u noassay a nd gas gr a phy ma ss spectrometr y . ig nificant correlation s of bl ood levels v with sal iva l e els were found for cocai ne and opiates .

These stud ies provide th e scientific basis for development of new non - invasive saliva tests for drugs of abuse . PUBLICATIONS

w M . J . L . . A . r E . J K K . v W k M ato , , Hillsg ro e , , einhold , , Gorelic , D , Da i n . and Cone , Coca ine and etabolite

- - S v U S N S . . A na l . 1 7 : 338 Excretion in ali a nder ti mulated and on ti mulated Conditions J Toxicol 341 1 993 .

S S S v A . N N E. J . 2 Cone , cientists tudy ali a Testing for Drugs of buse Employment Testing ; o . 1 1 , ov . , 93 .

- . 694: 91 1 2 99 S A . A . NY A S . 7 1 E . J . 3 . Cone , aliva Testing for Drugs of buse n n cad ci , SUMMARY OF WORK

r The pri ma y purpose of th is research is to assess the conseq uences of omg d ependence . Drug dependence ri v a d is thoug ht to have delete ous effects on the individual , produci ng se ere social , behavioral , n medical f rt t m consequences; however , ef o s to quantify these effec s have been hampered by a lack of i nfor ation about

ind ivid ua l i r . A rv the pr or to d ug use s a result , it is often impossi ble to deter mi ne if an obse ed cli nical condition is th e result of drug dependence or is reflective of a preexisting condition . Discordant identical , or monozygotic , twins offer a unique means for assessi ng the adverse consequences of d rug dependence . B ecause mem bers of monozygotic twins are genetically identical , the study of twi n pai rs in which only one twin is drug dependent provides a powerfu l assessment of th e effects of drug a buse whil e controlling for genetic varia bi lity . A secondary pu rpose of the study is the assessment of early experiential d ifferences (both d rug - related and non -drug related) to aid in the identification of non -shared environ mental factors important in the development of d rug dependence .

Through a larger twin study of th e genetic influences on d rug dependence conducted col la boratively by ARC r J k U investigato s and ohns Hop ins niversity , are identifying twi n pairs discordant for drug d ependence will ing rt - to pa ici pate in the ARC Discordant Twi n Study . T he d rug a busing mem ber of each twin pair is housed at the ARC residentia l unit for a 3-week drug - free period prior to study in or der to disting uish between acute

- u . A effects and long term consequences of dr g use fter this period , a va r i ety of factors associated with drug : dependence are assessed including medical disorders and various metabolic and ca rdiovascular effects , i personality and psych iatr c status , neuropsychological perform ance , and early environ mental experiences . With in - pair compa ri sons i n each of these domains will sign ificantly increase our knowledge concern ing

r - etiolog ical facto s i mportant in d rug dependence as well as the effects of long term drug a buse . PUBLICATIO N S SUMMARY OF WORK

T l r e here is sig nifica nt comorbidity between a cohol , d ug and mental d isord rs i n treatment sa mples a nd the

- n . T . co th ese disord ers general popu latio h e basis of this phenomenon is not clear The occurence of _ could be due to a common u ndertying genetic vulnerabi lity or it could be the r esu lt of one disorder se rving as a n _ environ mental influence that predisposes an individual to the other disorders . Th e elucidation of the etiology b rt of the comor idity wou ld be impo ant for prevention .

- r Twin cross conco da nt data provid e a method for assessing the cause of observed comorbidity . Twi ns are

- cross concordant if on e twin ,is affected with one disorder and the other twi n is affected with a second ‘ enetic fa ctor s - . I disorder f there are g com mon to both disorders , identica l twins would be cross concordant f more o ten than would fraternal twins . I f a significant difference is not found between identical and fraternal w v t ins , the observed relationship between disorders is likely d ue to en ironmental factors .

I - F r - nterpreting cross concordance data is made difficult by two factors . i st , cross concordance may reflect S the independent concordance for the second disorder that may exist withi n the twin pair . econdly , cross concordance may be th e result of an environmental association between the two disorders in the cotwin .

Data oh -comor bid drug and menta l disorders in tw ins with alcoholism will be com bined with appropriate control data in o rder to explore methods of analyzing the etiology of the association between these 1 69 - disorders . Data are availa ble on same sex twin pairs a nd analyses will ra nge from si mple cross ri - - concordance compa sons to full bivariate cross twin , cross trait analysis . PU BLI CATI O NS SUMMARY OF WORK

F ex osur e h a s b rt n neu r obeh a vior etal coca i ne p \ een repo ed to have effects on newbor , sleep and state n - d l EG . rl regu ation , and E patter s Ea y mother infant interactions have a lso been shown to be d isru pte among drug exposed dyads . The purpose of th is study was to investigate C N S development i n i nfa nts exposed to cocai ne i n utero and th e contri bution of variation in matern al interactions to this d evelopm ent . B oth cardiac

EEG N S 40 48 56 k . M and i nd ices of C development were assessed at conceptional ages of , , a nd wee s other rv 48 56 k infant interactions were obse ed at and wee s . Mothers and their i nfants were recruited from area

- full term newborn n urseries based on toxicology and self repo rts of cocai ne use during pregnancy .

Newborns were tested for cardiac reactivity to an auditory stimulus whi le still in the n ursery . I nfant cardiac EG W M - and sleep E ere then assessed at the specified ages . other infant interactions were videota ped d uring - - t I ma ter na l a face to face play interac ion and a feeding . nfant reactivity and temperam ent as wel l as psychopathology were assessed .

Analyses of the newborn data revealed that th e cocaine-exposed responded with slower ha bituation of behavioral responses a nd greater heart rate increase to th e fi rst auditory stim ul us presentation tha n the non T - r rv exposed . he cocaine exposed cried more often , experienced more state changes , had sho ter inte als of T rt contin uous sleep , and exhi bited evidence of autonomic instability . hese findings suppo previous studies suggesti ng poorer ha bituation among cocaine-exposed neonates and provide new evidence of altered t EG autonom ic reac ivity to environmental stimuli during the newborn period . Analyses of the E recordings -u wel l v S collected at follow p visits revealed de eloped sleep state organ ization in both grou ps . leep state v l M hea rt rate and aga tone did not differ between groups . others of cocai ne exposed i nfants tended (p 56 - - s to rate their week infants as more irritable and easi ly frustrated . During 48 week face to face i nteraction - - M - cocai ne exposed infants vocalized more than non exposed . others of cocaine exposed used more rs infa ntized behavio and responded more contingently to their infants than control moth ers at 56 weeks . PU BLICATIONS

SUMMARY OF WOR K

A T mong the most u biq uitous effects of addictive d rugs , but the least well studied , is d rug cravi ng . here has not even existed a validated psychometric instru ment for the evaluation of craving that wou ld l end itself to T A the systematic developm ent of medications for treating drug craving . he ddiction Research Center is presently developing such i nstru mentation for cocaine , h eroin , alcohol , and other drugs of a buse . Th is research shou ld he l p us to identify mechanisms of addictive action which are common across d ru g classes -oi w O and therefore lead to a better understanding the mechan isms hich underlie the drug cravings . ur initia l v v t findings i ndicated that cocaine craving is a multid imensional construct , in ol ing an ad m ix ure of urges and v A desire , intent to use , loss of control o er use , and anticipation of positive outcome . si m ilar ' mu ltidimensiona l str uctu r e was fou nd among responses from su bjects evaluated using the heroin

. O A instru ment ne hu ndred sixty su bjects have completed the new lcohol Craving Questionnaire . Th e project was modified in J u ne 1 993 to include measurement of craving and psych omotor/cognitive testing for outpatient resea rch vol u nteers and those on the residential u nit as a secondary study to exa mine th e t relationshi p be ween d rug cravings and human performance . Th e M arijuana Craving Q uestion nai re is u nder development for adm in istration I n FY 1 995 . Th e uti lity of computer administration of the series wil l be

. F l assessed to i m prove the efficiency of testing i na ly , to extend the generality of the research and S S investigate ethn ic and cultural factors , a tandard panish version of the Cocain e Cravi ng Q uesti on naires Ma nua l and has also been developed .

These adva nces in instru ment development and the enhanced under standing of craving should ena ble more rapid progress i n the a bil ity to produce more selective and efficacious medications and other interventions to meet th e needs of those addicted to drugs . With these instru ments we should be a bl e to predict wh ich craving d imensions may be relieved by medications and which will req uire other intervention to ena ble the person to ach ieve and sustain d rug a bstinence . PUBLI CATIONS

T . E . G . H a e rtze n C A . H enn in fi l S . S d v v e J . A . r Tiffany , , ingleton and g , Th e de elopment of a cocai n e c a i ng 34: 1 9-28 99 . A 1 3 . question naire Drug and lcohol Dependence , , SUMMARY OF WORK

Althoug h ea r ly ch a nges which occur du ring the smoking of marijuana are more li kely to be ind icative of its l r mode of action , the smoki ng phase of ma rijuana use has argely been ignored and ve y little is known w regard ing hat happens to a human su bject during the ea r ly phase of smoking .

k ri r n This study detai led the effects of smo ing ma j uana cigarettes on a va i ety of additio , blood a nd saliva m ri B v - u levels were deter ined d u ng and after smoking . lood and sali a l eve l s were com pared to drug i nd ced effects and horm onal changes .

' The resu lts fr om this study pro vide a comprehensive assessment of marijuana 's effects that occur both f during and a ter smoking and offer important insight to the mode of action of this widely a bused d r ug . PUBLI CATIONS

r l n . M . A . U M M ! Cone , E . J . and H uestis , , Do Consecutive i ne Catches Differ arijuana etabol ite Concentration J - 1 7 : 1 86 1 87 1 993 . Anal . Toxicol ,

H M A M E . J . L F M n I uestis , . . , itchell , a nd Cone , oweri ng the ederally andated Canna bi oid m m u noassay S S I M . . 40: Cutoff Concentration ncreases ensitivity and Efficiency , with inor Effects on pecificity Cli n C hem . ,

- 729 733 , 1 994 . SUMMARY OF WOR K

The dopam ine transporter has been identified as the pr i ncipal brain receptor site best corr elated with the ri rt - rewarding and eu pho c prope ies of cocaine . We have contin ued studies usi ng site d irected mutagenesis s r k a l T and molecular modelli ng and have pur ued wo with ph a nn co ogic structur e/a ctivity stud ies . hese joint strateg ies ai m to identify specific amino acids within the dopa mine transpo rt er that are m ore im po rt ant for cocaine analog binding than for dopamine u ptake , and to identify compou nds that could interact with these - I n v regions to provide dopamine sparing cocaine antagonists . this year we ha e further d efi ned the nature of rt W the interaction of cocai ne with th e dopami ne transpo er . e have used these data to defi n e smal l molecule lead compou nds that m ight interact with these transporter reg ions to provide dopamin e- spa ri ng coca i ne com lete U antagon ism . M ore p nderstanding of the interactions of dopam ine and cocai ne with the dopam i ne transpo rt er should lead to the el ucidation of better pharmacological agents useful in the treatm ent of cocaine abuse . PUBLI CATIONS

K S W - v M PP + S B U h l . itayama , ang J , G R Dopamine transporter mutants selecti ely en hance Transport . ynapse -62

- - S K W J B Y S A M K H M H a nd l er JS U h l . S urratt C , a ng , u hasz , mzel , won , , G R odiu m and chlorid e dependent

- k : cDN A . I : H S transpo rt ers in brai n , id ney , and g ut lessons from cloning and structure fu nction studies n ebert C

S . P v : S Gull a ns R , eds . Current opinion i n nephrology and hypertension ennsyl ania Cu rrent cience ,

60.

F M S S N T U T oh a ma M . O rt m R N A uuta , h imada , ishi mura , hl G , y ntogeny of dopamin e transpo er expression in - M B 6 . the rat brai n , ol rain Res

C holine/o ha n V8-2- 1 / rt mR N A AM h l G . r rv Gonzalez , U R p creati ne transpo er is expressed i n ne ous , renal and M B - 70. gastroi ntesti nal systems , ol rain Res

B JL A ku nne H C S ilverth om M L r Fl K B Pa rt il l a JS W J B Rothman R , Cadet , , , Car oll , Rice C , de Costa R , , ang ,

V . U G G J M . S . I hl owa R , Dersch C tud ies of the biogenic amine transporters Demonstration of a mu ltiplicity of , l i i i i 1 T 1 -55 J F T - b nd ng s tes n rat ca udate membranes for the cocaine analog [1 25 1R , E 309 .

K S U l G . P r rt J itayama , Doh i T , h R ho bol esters alter fun ctions of the expressed dopam i ne tra nspo er, Eu r - Pharm acol 9 .

i rt rt fa mil es Ex U l G J PS . N : J h R , ohnson eu rotransm tter transpo ers three i m po ant gene for n eu ronal fu nction , p

B iol l 9g4; in press .

S W P A M U hl G S B - chindler C , ersico , G R , old berg R . ehaviora l assessment of h ig h dose a m pheta m i ne

i rt i P rm B 1 994 in . wrthd r a wa l : m po ance of tra n i ng and testing conditions , h a acol B iochem ehav ; press

PUBLI CATIONS

- J a vo -A i h A i B e Voza r i S M P P d F U l d Y . lanchard V , R isman R , Vyas , ichel , y g , G , g Differential expression of tyrosin e hydroxylase and mem brane dopamine transporter genes i n su bpopu l ations of dopam i nergic neurons of

- B 38 . the rat mesencephalon , M ol rain Res

P t N S U K MJ . t m R N A Cerruti C , ilot e , h l G , uhar Reduction in dopami ne transpor er after cessation of repeated

- t M B 8 . coca in e admi n is ration , ol rain Res

AM W P A U . S Gonzal ez , alther D , azos , hl G R ynaptic vesicular monoamine transporter expression : distri bution

- h a rrn a col o ic M B 26 . and p g profile , ol rain Res

- t ‘ U W M F B Ja vo A id F . r M NA P k hl G R , alth er D , ash D , aucheux y g Dopamine transpo er essenger R in ar inson s A N - 8 . disease and control su bsta ntia nigra neurons , nn eurol

i M F A Kr E G 8 U hl G K J . reed CR , Revay R , Va ughan R , ek , rant , R , uhar Dopam in e tra nspo rt er i m m u noreactivity

J 1 994 in . in rat brai n , Com p N eurol ; press SUMMARY OF WOR K

The dopam ine tr a nsport er/coca ine receptor (DAT) is the site at wh ich cocain e experts r ewa r d ing/r e infor cing centr al effects and plays a role in termination of dopamine neurotransmission . This gene is expressed most

' ‘ prom inently i n just th ose dopaminergic neurons most impl icated i n cocaine s psychom otor stimir l a nt effects . ' We have isolated genomic clones of the h uma n and mouse DAT g enes that contain the -5 flan king s v in v seq uence , and ha e begu n work to identify the promoter for this g ene , order to pro ide means for ove r ex r ession k selective p of genes in just these dopaminergic ne u ronal popu lations . T h is wor also allows identification and study of new gene polymor h pisms that can serve as gene m a rkers to en hance power of A K t genetic analysis of D T roles in neuropsych iatric disorders . nowledge of th e promoter and gene s ructure - will all ow transgen ic m ice that-h ave mod ified dopamin e transporters or mod ified expression of the genetic repert oire of dopa minergic neuronsto be studied as potential a nima l models for hu ma n sub stance a bu se disorders .

N m early complete h uman gene and partial ouse gene sequences have been id entified a nd characterized . A short hu man DAT promoter seq uence appear s to d r ive expression of a repo rt er gene i n dopa m inergic

- neurons as well as n on dopam i nergic neurons of the brai n . PUBLI CATIONS

J S 8 8 U h rs . l . p Pe ico A M , Vanden bergh D , m ith G R Dopamin e transporter gene olymorph isms are not ass ociated with polysu bst ance a buse B iol Psychiatry - 7

' O H a r a B F DM L l B k M k M A KI a u nbe r S , Donova n indberg , ran noc T , Ric er D D . offatt C , g ch i ndler C , Chang P r oen ke h a l in S K N R J U . : T , elson , hl G R p transgenic mice a short promoter confers high testis expression and

- rt M R e r od 84 . reduced fe ility , ol p Dev SUMMARY OF WOR K

T l a nima ls i a nd here are arg e individual differences among humans and in behavioral , phys ological to in i i toxicological responses d r ugs of abuse . Many of these d v d ua l differences in behavioral responses to roa drugs display su bstantial genetic components . Transgenic animals provide means for app ch ing three interrelated g oa ls : 1 ) Identification of gene elements that confer cell-type specific expression and may th us allow targeti ng of i ntrod uced genetic ma ter ia l to appropr iate brain regions; 2) Elucidati on of gene elements yielding tra ns-synaptic gen e regulation and thus allowing appropriate regu lated expression of i ntrod uced genetic material ; and 3) Ascertainment of biochemical and behavioral consequences of the i ntroduction of or disruption of specific genes .

' centr a l v v Dopaminergic systems i nvolvement in mechanisms of reward and rei nforcement , and in ol ement of pre and post-synaptic opioid peptide systems in th e effects of opiate dru gs has led to focus on th ese ' ' systems du ring this FY . Elem ents i n the dopamine transporter gene s 5 flanking region that m ig ht confer its -s ecific 4 k e exquisite dopa mine cel l p expression were sought by cloning more than 2 b of this seq uenc , and d cel ls examin ing expression i n ifferent cultured and in transgenic animals . Tyrosine hydroxylase promoter , -s ecific which can provide catecholamine p gene expression , was used to drive expression of dopa m ine rt e P transporter, interesting transpo er mutants , and mu opiate receptor DN A s . relimi nary analyses appea r to mR N A reveal brain expression of but less expression of protein , while initial behavioral studies indicate a rt over ex r ession - varia ble i nfluence of transpo er p on exploratory , habituation , and cocai ne responsive behaviors . PUBLI CATIONS

U M r i r r oe nke h a l in hl G R , Takem ura . G ene mechanisms in p ma y afferent p p regulation i n nucleus caudalis .

i na a Y T oh a ma M . P . l nzl noki R , S h ge g , y , eds rocessing and inh i bition of nociceptive inform ation The

- 8 . V . N S B . etherlands: Elsevier cience P ublishers ,

U r i : I n : hl G R . A review of in situ hyb dization techniques relevance to com bined im munocytochem ical studies . 1 993 9 - A I . : W S S 1 3 00. Cuello C , ed . mmu noh istochemistry England J ohn iley ons , ; econd Edition , C hapter , 28

- - M AJ U . S c B arota J , Crosby G , hl G R elective effects of pentobarbital and halothane on fos and jun gene - 1 7 . expression in rat brain , Anesthesiology

FO S W M U . t alther D , Takem ura , h l G family mem ber changes in nucleus caudalis neurons af er pri mary

- - : B c M B 9 . afferent stim ulation enhancement of fos and fos , ol rain Res

U G I l L L u . I n : . hl R . dentify ing and oca lizing gene expression i mportant for th e actions of a bused dr gs ondon ed

- . B a : P 404 . I magi ng d ru g action i n th e brain oc Raton CRC ress ,

’ a r a B F UM L I B M M A KI a u nber BA S O H , Donova n , ind berg , rannock T , Ricker DD , offatt C , g , chind ler C , Chang

S K N elson R J U hl G . Pr oenke h a lin : r T , , R p transgen ic mice a short promoter confe s high testis expression and rt M R e r od - reduced fe ility , ol p Dev 84 .

h l P r e r oe n l r G M JJA U . ke h a lin s ina Crosby , arota , G R p p gene expression i n rat p co d followi ng su barachnoid 9 n analgesia with morphine or clon idine , Anesthesiology 1 93 ; i press . SUMMARY OF WORK

Psychostim u lants and O piates are know to induce behavioral dependence syndromes that are presu med to r o - m A be mediated by cor esp nding long ter changes in the central nervous system . lterations i n protei n / synthesis and or gene expression are strong candidate mechanisms to play rol es in th ese processes . Work in this laboratory during previous FYs has documented that psychostimulants and opiates can m odify the N - H A . expression of several D binding transcription factors in specific brain regions owever , which genes are i k regulated by those transcr ption factors remain largely un nown . Differential display foll owing polym erase chain reaction am plification (PCR - DD) provides a powerful n ew tool to identify such genes whose expression

evels . Y v v l are altered by d rugs this F , we have identified interesting candidate genes for in ol em ent ‘ During t in dependence mechan isms hrough this approach . PUBLI CATIONS

' l i P A M S W O H a r a B F B k M U h . B : ersico , ch indl er C , , ran noc T , G R rai n transcr ption factor expression effects of M ol B 1 99 0z91 -1 00 acute a nd chronic am phetamine and injection stress , rain Res 3 ;2

P A S W Z k B k M U hl . B r er sico , ch indler C , acze R , rannoc T , G R rai n transc i ption factor gen e expressi on , neurotra nsm itter levels and novelty response behavio rs : Alterations d u ring rat ampheta mi n e withdrawal and 1 94 i S 9 n . following ch ron ic i njection expression , ynapse ; press

PUBLI CATIONS

H S n e r S I - t i S h i e b . O eid breder Ch , G old berg R , p p g T n hibition of cocaine induced sensitiza ion by th e delta p oid

- na ltr ind ol e m 7 1 . receptor antagonist , Eur J P har acol

U 9593 m h i en ber S . 6 Heid breder C h , S pp g T prevents cocaine sensitization by nor alizing basal accu m bens

r e or t 1 994 . dopami ne , N eu r o p ; in press

S hi en ber S . S N I A Heidbreder C h , pp g T ensitization to the conditioned rewarding effects of cocain e , D Research

M onograph ; in press .

H S hi e n ber S . H eidbreder Ch , pp g T igh basal dopam ine tone in the n ucleus accu mbens blu nts th e response I n : L u ilot - . o L . I n profile to repeated coca ine exposure , Cador M , eds vivo methods for th e mon itoring of

B l nserm e 1 994 . molecules , ordea ux , ; in press

h i enber - S A E S T El G . M r chutz C , m brosio , pp g S , mer , H eidbreder Ch o ph ine ind uced locomotor activity and l r w L F : . I : L o o L dopami ne ove flo i n th e n ucleus accum bens i n ewis and ischer rats a comparative study n u i t , M - B er r n e 1 994 I n i l ns . Cador , eds . vivo methods for the monitor ng of molecu les , ordeaux , ; i n press

- i enber TS B a Is K . I mesolimbic S h ipp g , u bik R nvolvement of the dopam ine system i n m ed ating th e aversive P l 94 effects of opioid antagonists in the rat , Behav harmaco 1 9 ; in press .

enbe TS . l l O S hipp rg , Heid bred er Ch Ro e of kappa and de ta pioid systems in modu lati ng sensitization to the P rewarding effects of coca ine , Reg eptides 1 994; i n press .

e nber TS . S : S hipp g , Heid breder C h ensitization to the conditioned rewarding effects of cocai ne pharmacological J P m and tem poral aspects , har acol Exp Therap 1 994; in press .

nber TS . K S hippe g , Heid breder , Ch appa opioid receptor agonists prevent sensitization to the reward ing effects

. N I A M 4 of coca i ne D Resea rch onograph , 1 99 : in press .

enber TS S a na el M S hipp g , p g R , Heidbreder C h . od ulation of mesolimbic dopa m ine release by endogenous l ru - - r o e I : L ou ilot L M . l n opioids; in d g i nd uced sensitization and dependence . n , Cador , eds vivo m ethods for the r l a l nse r rn e 1 994 . monito i ng of mo ecul es , B orde ux , ; i n press

- a ib M S h i en ber TS . S ho , pp g Contrasting effects of adrenalectomy on n icotine ind uced d opa mi ne release and

- . I : L ouilot L M I n i locomotor depression i n rats n , Cador , eds . vivo methods for the mon itor ng of molecules , e 1 994 B I nsemi . ordeaux , ; i n press

a ib M S a na e l S S h i S S S ho enber . ; p g R , tohr T , pp g T train difference i n th e rewarding and dopa m i n e releasi ng P 1 994 effects of morphi ne i n rats , sychopharmacology ; in press .

a na el A O FX S hi enber S L - - m es l i b S p g R , l media , pp g T . ong lasting cha nges in morph in e ind uced o m ic dopa mine t S - 5 release af er chronic m orph ine exposure , ynapse .

S a na el A O FX B rt S hi enber S k : p g R , l meida a l C , pp g T . Endogenous appa opioid systems in opiate withdrawal mesolimbic P role i n aversion and accompanyi ng changes in d opam ine release , sychopharmacology

- 7 .

T - S a na el S hi enber S . v bina ltor im ine B N p g R , pp g E idence that nor ph (nor I) can fu nction as a n antagon ist at r ece tor t es J P m 1 994 multiple opioid p yp , Eur har acol ; in press . SUMMARY OF WOR K

The effects of drugs on pupil size are among the most easily detected and characteristi c sig ns of drug S l a nima l A ri action . evera studies conducted at the RC have cla fied the neu ral mechan isms through which i l psychoactive d rugs i nfl uence pu pi l diameter and the light refl ex. Th is research has been extended to clin ca stud ies where the pupil la yr effects of several classes of abused drugs were com pared to their performance and su bjective effects . Additional research on r etina l physiology using newly developed pu pil l ometers has yielded new i nformation of the retinal processing of the light reflex . S ince reti nal neural organ ization m imi cs u brai n neural systems , it is proposed that the influence of dr gs in this system indicates drug mecha nisms

l th e br a in . l e sewhere i n Dependent measures of these stud ies i nclude pu pi l size , constriction a nd di ati on _ s velociti es of the light refl ex , mooth pursuit , and saccadic tracking . These studies are typically within su bject u repeated measu re desig n , and d r gs are adm inistered in a double bl ind and placebo control led desig n ' P to d a te v rogress incl udes studies of the effects of marijuana , ethanol , cocaine , opiates ampheta min e ha e ri been st udies after va ous routes of admi nistration . PUBLI CATIONS

t S . E . B . r W . B . i J . B k k Fosnaug h , un er , , P ic wo h , Daily var ation and effects of am bient light and circad ian factors

- . M F P 553 992 . on the h u ma n light reflex ethods and ind ings in Cl inical and Exp eri mental harmacology , 1

X . Y . . . B . M G k . A . G P k T K I . R . B . W . B . Rothma n , , orelic , D , uo , , H em ing , R ic worth , Gendron , , oeppl , , Thomson

- - E . H en ni fl eld J . L k i I l l n . L . E. , and g , ac of evidence for context dependent cocaine induced sens tization

P harm acology B iochemistr y and B ehavior 1 994 (in press) .

A J . P J R E . k B E . S v i W . B . h s s ol o ic ickworth , , oh nson , H olic y , , and Cone u bjecti e and p y y g effects of Cl in ica l -576 . P 1 993 . bupren orphin e in humans harmacology and Therapeuti cs , SUMMARY OF WOR K

A major research emphasis i n the last year has been to uti lize the behavior genetics method and a classical pharmacological approach to determ ine genetic and pharm acological contr i butions to characteri sti cs - e rt . I n impo ant in th e acute effects of opioids particular, delta opioid subtypes have recently b en proposed

' ba sed u on - DPDPE p d ifferential antagonism and no cross tolerance between the delta agon ists , a nd

- deltor phin I I . I n order to determ ine the existance of delta receptor su btypes and determ ine the deg ree of genetic covariance between the proposed subtypes full dose- response curv es for DPDPE and d e ltor ph in I I P were determi ned in eight in bred mouse strains . The correlation between sensitivity to DPD E and deltor phin l I - ind uced analgesia was not significant thus suppo rt i ng a pharmacological a nd genetic separation

- - of the two delta receptor su btypes . Two in bred strains that lacked a delta 2 response and a strain in wh ich the mu agon ist morphi ne acts like a delta -agonist were used in com bination with other i nbred strai ns to -2 - - - investigate the interactions of the delta su btype with delta 1 and mu agonists . Cross over pretreatment experiments with the two delta -agonists suggest that deltor p h in I I is not u niq ue in its pharmacology d ue to rt I n partia l agon ist prope ies but is pharmacologically and genetically distinct in its mechan ism of action . the 2 l - interaction experiments , th e strains lacking de ta 2 agon ist effects and the strain i n wh ich morph in e acts via - d eltor hin I I - I delta m echanisms , p did not shift the morphine dose response cu rve . nitial biochem ical studies done i n colla boration with the Clin ical Psychopharmacology S ection verify the a bsence of a delta - receptor subtype in the C BA strain and may provide a unique 'd knockout' strai n for in vestigating th e r ole of this d r ece tor su bt e r ovied n p yp in the acute and ch ron ic effects of a bused drugs . Th e data p i these experi ments are being com pared to transgen ic mice that display increased mu-receptor B ma x (superoxide d ism utase transgene addition , M olecular N europsychiatry Section) in order to explore the i n vivo conseq uences of altered a nd delta receptor reg u lation on the acute and chronic affects of a bused d rugs PUBLI CATIONS

f th r - B ri S k v 8 S . o otr o in o sova E , uda o , El mer G I , Goldberg R Action y p releasing hormone on m orphine induced P B B v 1 994 analgesia a nd preference in two in bred rat strai ns , harmacology , iochemistry and eha ior ; in press .

F v I . : Elmer G , George R Rate depressant effects of ethanol in selecti ely bred mice relati onsh i p to acute J A t - u r sensitivit 1 9 . ne o y to ethanol , ou rnal of ddic ive Diseases

G I F A - i R O 5 45 1 3 . 1 A : Elmer , G eorge R ntagonism of ethanol induced narcos s by a nd indom ethacin lcoholism 1 994 Clinica l a nd Exper i m ental Research ; in press .

- G G I Y H YT L H B . M a ntio i i Goodma n C , Elmer , ang , ee C , Rothman R odulation of opioid receptors by p o d m L F . P A I : rw P 1 994 . peptides . n Tseng , ed ha r acology of opioid peptides , Ha ood cademy u blish ing , ; in press

M M G I G S . h a rrn a co enetic a r l ey J , Elmer , old berg R The use of p g tech niqu es i n drug a buse research , P T -37 harm acology and herapeutics .

M M S K Elmer I M L arl ey J , himosato , G , iner L . P harmacogenetic approaches to d rug dependence . I n :

- W S L GG . N P P L 1 993 9 onnacott , unt , eds eurochemistry of d rug dependence , ortland ress , ondon ; 5 z1 53 72 . SUMMARY OF WORK

The primary focus of th is research is to develop a better understanding of the pharm acological mechan isms u nderl yint h e behavi oral effects of cocaine that lead to its a buse and th e conseq uences of that S : 1 T a s' indica ted abuse . tudies have i nd icated that ( ) he psychomotor sti mulant effects of cocaine , by

D 1 D . increases in locomotor activity , are mediated by both and Z dopam ine receptors For increase in v DZ learned operant behavior produced by cocaine , howe er, receptors have a greater i nvolvement than do T v 2 D 1 receptors . he respective roles of other dopamine receptors are cu rrently under in estigation . ( ) There are differences among dopami ne upta ke inhi bito rs in the relationsh ips of their bea vior a l effects a nd their affinity for the dopamin e transport er . For drugs that have a str uctu r a l similarity to coca ine there is a d i rect - rt v v I n relation between affin ity for the transpo er and in i o potency . contrast , for structu ral ly dissim ilar compounds their is no simple relationship . These data indicate that cocain e and its congeners bind to the k 3 dopam ine transporter i n a manner that is disti nct from that of other dopam ine u pta e i n hi bitors . ( ) The “ m D subjective behavioral effects of cocaine are ediated by both 1 and 0 2 dopamin e receptor systems , a lthough actions through either system alone are not sufficient to fully r epord u ce the su bjective effects of 4 t cocaine . ( ) The su bjec ive behavioral effects of l ow doses of cocaine are mediated by both D 1 and DZ

D l . I n dopam ine receptor systems , although actions through the dopamine system a ppea r to predom inate addition , the su bjective effects of low doses of coca ine have a significant noradrenerg ic compon ent that is 5 nh not evident in the effects of higher doses of cocaine . ( ) A ydr oecgonine methyl ester (A EM E) is a major pyrolysis product of cocaine and has been detected in high concentration in the urine of su bjects who have " - . I n A EME stru ctu r l l a r esembl ence A . smoked crack add ition , has a to anatoxi n , a known cho in erg ic toxin ri A EME " T The cont bution of to the u nique pharmacological actions of crack was i nvestigated . hese studies found that A EME probably does not contribute in a significant way to the stimu lant or su bjective effects of 6 crack or its toxicity . ( ) Tolerance to the behavioral effects of cocai ne is not accompa nied by changes i n l rt the fu nction of dopam i ne D receptors . Fu her studies are being conducted to assess th e effects of cocai ne on functiona l 7 U treatment _ aspects of other dopamine receptor su btypes . ( ) niq ue com pou nds have been synthesized that have h ig h affi nity for the dopami ne transporter and in hi bit dopam ine u ptake . However, I - these d rugs do not have behavioral effects that are similar to those of cocaine . nitia l stru cture activity studies ind icate that the binding of these compounds to the dopamine transporter is different from that for

- cocaine . B ecause these compounds bind to the dopamine transporter but do not have cocai ne l ike t r v behavioral effec s , they may serve as cocaine antagonists , wh ich is cu rently under in estigation . PU BLICATIONS

k A N H S S W i Erzou i H K A A . , llen C , ewman , Goldberg R , chindler C Effects of coca ne , Coca ine m etabolites and i r - b cocaine pyrolysis products on the hindbra n cardiorespirato y centers of the ra bit , Journal of Card iovascular

I n 1 994 . P harm acology , P ress ,

“

wa sser S r P B W k J M K J L . lzen , Ter y , Heller , it i n , atz Differential relationships am ong dopam in e transpo rt er r i k P I P affi nities and stim ulant potencies of va ous upta e inhi bitors , European J ou rnal of harm acology , n ress

1 994.

l e nwa sse r 3 -d i h e n l m eth ox r o a ne N AH l l e n A z S K J L . N v t ewman , A C , , atz o el alpha p y y p a nalogs are potent k - k M dopam i ne upta e inhi bitors without cocaine li e behaviora l profiles , J ournal of edicinal Chem istry -91

N A A W J M lzenwa sser S M K J L T ewman H , Allen C , itki n , , ash D . atz . h e therma l decom position product of " A EME i crack , , a nd analogs do not a ppear to contr bute acutely to the pha rm acological or toxicol og ical actions Medicina l r - 1 1 of cocaine , Ch em ist y Research 0.

T P W J M K z J L . P i i err y , itki n , at harm acological character zation of the novel discr minative stim u lus effects of a w J n l P rm E I P 1 994 . lo dose of coca i ne , our a of ha acology and xperimental Th erapeutics , n ress ,

- i l l E J M . T rs f T re i , Witki n ransient hype ensitivity to apomorphine induced gnawing a ter term ination of acute

- l B P 98 . effects of a sing e hig h d oses of cocain e , ehaviou ral harm acology

Ph a mi a coth er a W k J M . P N B iobe h a it in py of cocai ne abuse : reclin ical development , eu roscience and vior a l -4 Reviews 2 .

PUBLICATIO NS SUMMARY O F‘ WO R K

The dopam inergic system is clea rly impl icated as important in mediating the effects of many d rugs of Pa rk ifisons O m abuse , as well as disease , and schizophrenia . ur studies have as their goals the deter ination w i a of the functional sign ificance of the dopam ine system in normal functioning , as well as ho t cts to su bserve dru g a buse . O ne specific objective is to better characterize the pharmacology of the va r ious subtypes o f C N S dopam i ne receptors . I ncluded in this goal is the identification of drugs that act selectively h and with h igh efficacy . I n many cases t e pharmacological tools for the study of these receptor su btypes in

. A r esu lt rt v r ew i wrl l vivo and i n vitro are lim ited s a , one fu her goal is the d isco ey of n synthetic ent ties that : 1 allow analysis of the pharmacology of these dopamine receptor subtypes . These studies ind icate that ( ) W f e D 1 e vi hile it is possi ble to d i fer ntiate dopami n agonists in tro on the basis of their intrinsic efficacy , “ differences i n the efficacy of the d rugs have not been correlated with any other observed pharmacological

e . 2 S S KF 38393 effect of thes drugs ( ) tudies of the behavioral effects of the D1 dopam ine agon ist , , have T indicated few of its behavioral effects are mediated by actions at D1 dopa mine receptors . his fi ndi ng is S KF 38393 D1 3 D1 important because is often used as a prototype agonist . ( ) Dopam in e , but not dopam ine DZ v v W , receptors are in ol ed in the l ethal effects of acutely administered cocaine . e a r e preparing D l antagonists that do not penetrate the blood brain barrier. These drugs may serve a s a ntid otes to acute cocaine overdose and wil l also serve as tools for the investigation of the fu ncti on of the peri ph era l 0 1 dopamine system . (4) Dopamine DZ agonists can stimulate this behavior in a manner si mi lar to the ma n ngr v a D1 . in which cocai ne or a mphetamine stimu l te beha ior, in contrast , agon ists do not _ These d ifferences a n DZ r suggest involvement of dopamine recepto s in the stimulant behavioral effects , a nd a lack of D 1 5 involvem ent of dopa mi ne receptors . ( ) The pharm acology of dopam ine DZ agonists is being character ized in a cell li ne that expresses a DZ receptor that is functional ly simil a r to brain dopami ne DZ

’

. B DZ receptors ecause th ese receptors are expressed in th e a bsence of other dopam i ne receptor su btypes , l a n T this cell i ne is a model system for studying the function d regulation of the DZ receptor . h e findings from these studies suggest that the dopamine DZ receptor regu lates cyclase in hi bition predom inantly via the Gil - and/or Gizsu bu nits of G alpha . Using th is system the intrinsic efficacies of dopamine DZ agonists have s been characterized . Th ese studie also indicated an intrigu ing link between the Gi3 su bu nit and the G il or

- Giz . 6 D3 -O H P T su bunits ( ) The pha rm acology of the putative receptor agonist , 7 D A , is being

- - characterized . These behavioral and in vitro studies have i ndicated that 7 O H DPA T is a weak pa rt ial DZ agonist at receptors , and its several of its effects may be mediated by that activity rather than DS agonist PU BLI CATI O NS

S K - - T i D2 - L se r z JL . 7 O H DPA lzenwa s , at antagonizes dopam ne receptor inh ibited adenylyl cyclase activity , ife

- Sciences 1 994; 55 : PL 2 57 9 .

S I D a se r . 2 : lzenw s , Cote TE nhi bition of adenylyl cyclase by a homogeneous population of dopami ne receptors S G i1 / G iz N r I P 1 elective blockade by a ntisera directed against and or , J ou rn al of eurochemist y , n ress , 994 .

J M A K W k K J L . P Pel l O n R , Flores P , ll ing , it in , atz harmacolog ical analysis of the scratch ing prod uced by DZ k J n P Ex e r im enta l I P dopam in e agonists in squi rrel mon eys , our al of harmacology and p Th erapeuti cs n ress ,

1 994 .

i - T e P K z J L . A r D 1 S H 23390 S H 39 6 ny , at compa son of the effects of receptor antagonists C and C 1 6 on D- 1 S KF 38393 P 1 994 1 1 3 z328- 3 suppression of feeding behavior by the agonist , sychopharmacology ; 3 .

Vertica liza tion E W J M . r Tirelli , itkin of behavior elicited by dopamine g ic mobi lization is q u alitatively different 7 6J DBA /ZJ w CS B L/ P I n P 1 994 . bet een and m ice , sychopharmacology , ress ,

J M . w Tirelli E , Witkin Differentiation bet een direct and indirect dopami ne agonists via their effects on gnawing C57B 1 /6J J n P m E I n P 1 9 4 in mice , our al of har acology and x perimental Therapeutics , ress , 9 .

- E J M . T s f m Tirelli , Witki n ransient hyper ensitivity to apomorphin e induced gnawing a ter ter i nation of acute B P - effects of a single hig h doses of coca ine , ehavioura l ha rmacology 98 . SUMMARY OF WOR K

A r . lmost al l of our knowledge of drug recepto s comes from i n vitro experiments However, it is i mportant to

‘ i o f v v or . A oa ls study receptors in many reasons ccordi ngly, one of our g is to develop liga nds and r P i approaches for studying d ug receptor sites in human populations by imaging techn iq ues . r or to i magi ng in n huma s , ligands need to be developed and tested in vitro and in vivo in animals . I - i R T l -55 n our stru cture activ ty studies of the cocaine receptor , was identified as a very potent com pound with a high affin ity for the dopam ine transporter . It a lso has some affinity for the serotoni n transporter as A h R T l -55 well . ccordingly , our previous pu bl ications s ow that is an excellent P ET and S P ECT liga nd for

. v . v studyi ng dopa m ine transporters in vi o I t has been patented , licensed and is cur rently being used in se eral e P ' centers as a n i maging reag nt for diagnosing arkinson s disease . B rt r I - 55 eca use of its significant affinity for serotonin transpo e s , it is possi ble to use RT as an in vivo rt s W binding ligand to study the occupancy of serotonin transpo er by relevant d nr gs . e have exa m ined the rt u impo ant n ewer antidepressant dr gs which have a selective affinity for th e serotoni n transporter . T I -55 fl uoxetine Radiola beled R was admi nistered and in vivo competition was ca rried out with , paroxetine

. A clea r and sertraline t behaviorally effective doses , it was that these d rugs occupied th e seroton i n

- rt . A k transpo er lso , fluoxetine which is nown to have a longer half l ife in hu man su bjects , had a m uch longer - a nima l I - half life in these studies than the other compounds . These results ind icate that RT 55 ca n be used to rt rt study the serotoni n transpo er, in pa icular to identify d rugs that bind to that site , to determi ne the relative rate of occu pancy , and also determ ine the duration of action of com pounds at this site . We have previously uti l ized radiolabeled WI N as a PET ligand to bind to dopam ine transporters in I n s . k vivo a recent study , we exam ined the inetic and pharm acology of this bi ndi ng in ani mals and h umans . We clearl y show that th is compound is a promisi ng radioligand for future studies of neuropsychiatri c disorders that involve th e dopamine transporter site . PUBLICATIONS

H K rt . T . C . A . I Y B na ls F S E . B F S W D . F . Da n R . . U . t ong ung , haya Rave , Chen , Chan olio cheffel , Ricaur e J I n N . . M . . N J W H . J K B . I G . A . L , eum eyer ag ner , r and uhar Vivo magi ng of a boon and H uman Dopam ine P U 1 1 C W I N S 1 5 1 30- 1 42 1 993 Transport ers by ositron Em ission Tomography sing [ 1 ynapse , , , .

J M . O J . S U K . K S rt S . E . . F cheffel im , Cl ine and uhar ccu pancy of the eroton in Transpo er by l uoxetin e .

- - P S rt : I n v S W 1 25 l R T l 55 . S 1 6 263 268 1 994 aroxetin e , and e raline Vi o tudies ith [ ] ynapse , , . SUMMARY OF WORK

The goa l of this pr oject is to expl ore the i nteraction of coca ine and cocai ne-li ke compou nds with th eir T he " binding sites i n brai n . binding site or recepto r thought impo rt ant for the reinforcing effects of cocaine , a nima ls mi O r at least in , is th e dopa n e transporter. ther effects of cocaine and its congen e s are li kely to be due to interaction at other sites such as other transpo rt ers Besides developing an u nd erstanding of the h rt l interaction of cocaine wit transpo ers , this study a lso reveals potent reversi b e and irreversi ble bi nding v S ligands for i n vitro and in vi o studies of transporter proteins . ome compou nds may be useful as treatment medications as wel l . S everal classes of cocaine analogs were tested in binding and upta ke studies at th e dopa mine transporter , a n r and someti mes lso at the orepineph i ne and serotonin transpo rt ers . These studies were carried out using T rat brai n tissue althoug h sometim e human brain tissue was utilized as well . he methods em ployed were l e standard radio a bel d ligand bi nd ing techniques . b - oxa d ia zoles Cocaine has a car omethoxy group in the C 2 position which is required for its activity . S ince bioisoste r es v oxa dia zoles are excellent of ester groups , se eral were synthesized and tested i n bi nd ing I n assays . general , th ese compounds showed potencies for the dopamine transporter si m ilar to the parent

- r . T l C 50 nM sel ectiveit este s he most potent analog had an of , and y for the dopami ne transporter was e ll high as w . These com pounds may have utility as medications to treat d rug a buse si nce replacement of w will - the ester ith the oxad iazole produce a compound that is resista nt to metabolism a nd long lasting . By produci ng a ser i es of compou nds without the methyl group attached to the nitrogen i n the ring structure we found that demethylation enhanced affinity for the serotonin and norepinephrin e tra nsporters between 2 - r r and 44 fold . This will be ve y useful when t y ing to design cocaine analogs selective for seroton i n a nd norepi nephrine transporters . A C-2 T series of amides in the position were synthesized and tested as well . h e results indicated th e rt S tertiary amides are more potent at the dopam ine transpo er than primary and seconda r y a m id es . ome of the compou nds were both potent and highly selective for the dopami ne transport er. A QSAR and CoMFA study extended knowledge from the str ucture-activity work and elucidated som e ha r rn a co hor e A features of the cocaine p p and provided useful predictive inform ation . dd itional com pounds l wil be synthesized using this information . PUBLICATI O NS

W . M J . . Kuze mko . . J . a sca r e l l a . . Y H L B K M A P A A . F . l . S . W . M . . Carroll , , , . Gao . braham ewin . oja . and uhar ' S L B SA CO M FA S 3 b - S P h e n i 3 b 4 ynthesis , igand inding , and Q R ( and Classical) tudy of u bstituted y ) a nd

- - r . . Disubstituted Ph e nyl)tr O pa ne Zb carboxylic Acid M ethyl Este s J of M ed . Chem in press

L . . . . Kuzemko . W . M J . . 3 b P K J . A H L B K P A M A . J . F l . . . Carrol l , . , otian , G ray , braham , ewin , oja , and u har (4C h lor oph enyl)tr op a n-Zb— Ca r b oxa m id es and Cocaine Amide Analogues : N ew H igh Affi n ity a nd S electi ve

- . M . . 3 468 47 1 993 . Compou nds for th e Dopam ine Transporter ed Chem Res . , 2 ,

K a tic J . o . . B . . K Y H L S W . M . . P I . J . P A A A oja , , uhar , T p j , E a ng , braham , ewin . a nd Carroll . econdary mine ' Analog ues of 3b-(4 S u bstituted ph enyl)tr opa ne-Zb— carboxylic Acid Esters and N - N orcocai ne Exh i bit Enhanced i - A S N rt rs . . M 37 1 220 1 223 1 994 . ffinity for erotoni n and orepineph r n e Tra nspo e J ed . Chem . , ,

- - L . . . Ku I . J . . zemko J F . G P A M A A L B K A J . W . M . . 3 Carroll , , ray , braham , , . H . ewin , oja , and uh ar ryl Z ' Substituted -oxadiazole-S yl)tr op a ne analogues of cocaine : A ffinities at the cocai n e bi nd ing site at the i rt J M 36 2886-2890 1 993 dopami ne , seroton in , and norepinephr ne transpo ers . . ed . Chem . , , .

PUBLICATIONS

S K J W N . S . L G . M . . m Pilotte , harp e and uhar ithdrawal of repeated inter ittent intravenous infusions of cocaine results i n the delayed reduction of binding to dopamine transporters in the n ucleus accum bens of Lewis Rats . - FE 269 963 969 1 994 . J T , , ,

M . P . U K C . N . S GR J . rt mR N A Cerr uti , ilotte , h l , and uhar Reduction in dopam ine transpo er after cessation of - . MO I . B 1 32 1 3 8 994 repeated cocai ne ad min istration rain Research , 22 , , 1 .

M . J . G W M . K U mR N A a mi . D . . R . I C ti , C alther , , uhar, , and hl , Dopam i ne Transporter Expression is ntense in M N O B 1 8 1 8 1 -1 86 M M . M . 1 993 . Rat id brain eurons and odest utside id brain ol rain Research , , , SUMMARY OF WOR K

The smoki ng route is an efficient means of d r ug delivery . N u merous d rugs of a buse are ad mi nistered by this ri M route incl uding cocaine , ma juana , phencyclidine , heroin and methamphetamin e . ethods were developed “ to study the pha rm acokineti cs and ph a rrn a codyna mics of smoked cocaine and heroi n i n h um a n volu nteer subjects . Healthy su bjects with a recent histor y of a buse of the d r ug of interest by the smoki ng route participated I n the stud ies . m l ow Following i nfor ed consent , the su bjects smoked a dose of the d rug u nder controll ed cond itions to

. n assess safety of the procedures The su bjects then participated in a double bl i d , placebo control led study of smoked versus intravenous adm i nistration of drug . B ehavioral and physiological. measures and biolog ical r samples were collected ove tim e . These data provide impo rt ant information to our u nderstanding of the d m s k pharmacologic actions of these g by the smo ing route . PUBLI CATI O NS

R M H en n in fie ld J E . J . Ph e k J . K . E . a r rn a cokin tics m A . A P Jen ins , , eenan , g and Cone , nd har acodynam ics O f

I 99 . S J A . n 1 4 moked Heroin . . nal . Toxicol P ress SUMMARY OF WORK

The Cli ni ca l Psychopharmacology Section conducts preclini ca l and clinica l research into the m echanisms of

t . A ma O r t N I DDK ac ion of cocaine j component of this project , conduc ed in col laboration with i nvestigators at and N I M H , is the synthesis and evaluation of analogs of G B R 1 2909 as putative coca ine a ntagonists or

- . A rt cocaine su bstitution type medications s pa of this project , we synthesized and eval uated the fi rst ch iral " " G B ri e na ntioselect k I t R de vatives as potent and ive inhi bito rs of dopam ine reupta e . n a n ex ension of - previous studies , we showed that daily administration of G B R 1 2909 suppresses cocain e self ad m in istration in Rhesus mon keys without the development of tolerance , supporti ng its potential use i n the treatment of coca i ne addiction and that G B R 1 2909 blocks the increase in extracellular DA produced by intraven ous cocai ne . Another com ponent involves investigation of the possi ble heterogeneity of DA transporter bind ing

. l a ffinti sites Th is project has id entified m ulti p e and novel binding sites for the h igh y cocaine analog , l - [1 25I] R T 55 . Another com ponent of th is project addresses the role of classical condition ing in cocain e induced behavioral sensitization . These studies demonstrated that associative learn ing mechan isms are -s ecific S involved in the acq uisition of context p behavioral sensitization to cocai ne . tudies with g enetical ly inbred strai ns of mice showed that the occurrence of sensitization is not correl ated with either th e potency or

- efficacy of cocaine as a motoric stimulant . Huma n studies failed to demonstrate cocaine sensitization with a one day tra ining parad igm (l R P Open - la bel studies with phenterm ine a nd fenfl ura m ine showed that these m edications suppress cocai ne craving in addicts seeking treatment . Clinical research protocols are in preparation to deter m ine the efficacy of fenfluramine and phenterm i ne as treatments for a lcohol cocaine addiction . PUBLI CATI O NS

l . r . M H S U B . . K . . F . B . A ku n ne . Pa rt il la . . . Dersch , C . . C . , J . , G Char , R d e Costa C Rice , Ca r ol l . and R

S . k k Roth man tudies of the biogenic amine transpo rters l . dopam ine reupta e bloc ers inh i bit [3Hjma zindol bi nding to the dopam ine transporter by a com p etitive mechan ism : prelim inary evidence for - c em R es 1 9 Z : 201 208 . N e u r o h . different bind ing domai ns . , ( )

I r . . R a d esca . . . . v K M L . B K F M B . Rothma n , R . B . , . . ecketts , . R , R de Costa , C Rice , y Ca roll , and C Dersch S i 3 H - - k - tud ies of the biogen ic amine transporte rs . I I . a br ef study on the use of [ ] DA u pta e in hi bition to

- - transpo rt er bi nd ing inhi bition ratios for the in vitro evaluation of putative cocaine antagon ists .

B M M B k B W A . . . . auman n , . H roc ington , and R Rothman ithdrawal from chron ic cocaine enhances

- - the 5 T 1 C 577 . v H Z/ O I . B . beha ioral sensitivity to agon ist D iol Psychiatry ,

m B M J J . S . A I H . . B . au man n , . , Rutter, and uerbach ntravenous admi n istration of th e serotoni n agon ist chlor oph enylpipera zine (mCPP) i ncreases extracellular seroton in in the diencephalon of awa ke rats . rrn a col o -1 386 N eu r oph a gy , .

A li P . S S . R . B . Ca ri son A J . . . Cadet , L , heng , , Rothman , E . , and C Epstei n tten uation of - N e e . co er/zinc . J . ur och m metham phetam ine ind uced neurotoxicity in pp superoxide dismutase transgen ic m ice , - 383 . 62(No .

C . . . rs e . M J . M B G U J S P K t e . . . u nn L . . . . . r B A k , H , C De ch , Cadet , H au mann , . Char , . a i l l , R de Costa , C . Rice ,

F . . S m r . I . . l . Ca rroll , a nd R B Rothma n tudies of the biogenic a ine transpo ters l l d em onstration of two

3H GB R 1 2935 H J P . 3 BT CP . . m . Ex . 268 binding sites for [ 1 a nd [ ] in rat ca udate mem branes har acol p Ther , -1 475

- H . . . M . J P . L S B E 5 HT Z/1 C Bauman n , , L Cadet , . heng , and R . . Rothman vidence for alterations i n N I A 1 993 . Mono r . . P receptor sensitivity after repeated cocaine injections roblems of Dru g Dependence D Res . g ,

m . k K . M M . S . S ilverth o J M S K L B L . . B . . . :l . . Pa rtilla . . ec etts , , Cadet , C . Dersch , , R de Costa , C Rice , - sca . l . . . o R a de , F Carrol l and R B R th man Resolution of two serotoni n transpo rt er related bi nd ing sites l 1 25 I T I - 55 I A . N . Mono r . with the cocai ne ana og , [ 1R D Res g ,

. . Gor c B k A . B D . S G I G el i k . roc ington , , R Roth man , , R . Gold berg , and . . Elmer enetic factors in acute ‘ - - 41 N I A M ono r 1 : 1 6 . cocaine induced locomotor activity and context specific cond itioned sensitization . D Res . g

J . L . T . W . . L J . B . H ennin fie ld . Cadet , , Gendron , R ang e , E g , and R . Rothman The cl i n ical n eurological i : N I A r . M ono . exam inat on in chronic cocai ne abusers prel im ina ry findings D Res . g ,

- . . l T . . U M B . . K B B . M a . . . . k Char , G H au man n , C Chen , C Rice , R . d e Costa , D . a tec , a nd R Roth ma n The cocaine- induced rise in extracell u lar dopamine in the n ucleus accum bens is reduced by G B R - 1 2909 M N A . . I ono r . pretreatment D Res g ,

C . . . . . Pa rtill a J J S . . K F l A e B . L B e A ku nn . Dersch , , Cadet , , R de Costa , . C . Ric , . l . Carrol , H . . , and R Roth man

- Demonstration of m u ltipl e binding sites in rat caudate m em branes for th e cocai ne a na log [1 251] R T l 55 .

M ono r . N I A . D Res g ,

G T . . B . J . L . B S K A G k J W B P . . n fi ld . . endron , , R Roth man , Cadet , oeppl , D . , orelic , . E . H enni g e , and ickworth

P N A M ono r . I . ossi ble cond ition i ng effects of intravenous cocaine on pupil la ry d iam eter . D Res g ,

. R . . . . M a G J F W B . K B T tecka . . lowa , , H ojnicki , de Costa , D , C Rice and R . . Roth man h e effects of 1 2909 G B R on respond ing of rhesus monkeys maintained u nder schedules of cocaine and food d el ive ry . N I DA

1 41 : 1 2 . Mono r . Res . g ,

k . . . Ma tec a D . B . B . M L i L M K M B k A S lve . . . , R de Costa , R Rothman , . . rt hom , . R a desca , C . . Dersch , ec etts , " P J . G K . . S G ert , lowa , and C Rice ynthesis and evaluation of the fi rst chir a l B R der ivatives as potent SUMMARY OF WOR K

The C N S synthesizes and secretes several neuropeptides which attenuate the actions of morph ine including CCK-8 T -M I F P -Le u - P -G I n- P -Gl n-A r -P - N HZ N PFF - M S 1 , yr , he he ro g he ( ) , al pha H and dynorph i n( The anti opioid mod el of tol erance and dependence postulates that ad mi nistration of morphine prod uces increased i - secret on of a nti opioids , which attenuate the effects of morp h ine , and thereby maintain a homeostatic

- - balance . A prediction of the anti opioid model is that administration of an anti opioid shou ld atten uate th e

- development of tol era nce and dependence . A major finding of this project is that adm inistration of a nti N PFF

- I G . I n g to dependent rats atten uates naloxone induced withdrawal addition , recent autoradiograph ic studies have shown that the d ensity of th e opioid mu receptor in the brain is u nder tonic inhi bitory control by N PFF . rt m u r mesol imbic F P N P F . l mpo antly , the recepto s in the system are regulated by reli mi nary stud ies indicate N PFF e that antagonizes the r inforcing effects of morphine . Heroin addicts suffer from dysphoric mood i i ri O states pr or to and du ring their addiction , as wel l as dur ng pe ods of absti nence . ne hypothesis to explain

this is increased levels of dynorph in , which , via activation of kappa opioid receptors , produces dysphoria . This hypothesis is bei ng tested i n the cl in ical protocol : Anti-opioid peptide levels in th e plasm a a nd CS P of drug a busers and age matched controls (l R P We continue our colla borations with m ed ici nal chemists to identify potent and selective kappa antagonists . The sig nificance of this project to drug a buse r ese l R Ph is that the d eli neation of novel mechan isms involved i n opioid tolerance and dependence wil l eventual ly lead l to nove , and more specific treatments for addiction . PUBLI CATIONS

B L B H B L I ntr a cer e b r ove ntncu l a r 8 . . Xu . . Roth ma n , R . . rady , , and J ong C hronic infusion of the anti P - L -P - - P - -A - P - opioid peptide , he eu he G in ro G in rg he down regulates m u opioid bindi ng sites in rat P - 1 277 brain . eptides ,

- - H . . C . H . . . G J L . H . B Y L B M Y . B . . . M oodman , C . , Cadet , aumann , T ang , ee , and R Roth man odulation - M on r N P FF . N A . o . of opioid mu receptor density by the endogenous anti opioid peptide , I D Res g ,

- P J Y . B . I O J . . a rtill e , , ou and R Roth man solation of piate and anti opiate peptides from hu ma n

M ono r . N A . plasma . I D Res g ,

A . K K . . B . . B L B . . . . Raffa , R . , i m , C Rice , R de Costa , E E . Codd , and R Roth man ow affi n ity of a mid e Pa R Ps FMR Fa mid e- - F- 8- F e FMR F and fou r ( related peptides) , including the mam malian d erived mide A - 1 8-Fa mide O k -404 PFF 1 kZa kZb . P . (N ) and , for pioid m , d , , or receptors eptides .

- l . G . G . G . H Y . Y B M . . . L . . oodma n , C , Elmer, T ang , C H ee , and R Roth ma n od ulation of opioid - . I n P r O P H a n/vood A P rs G M B receptors by anti opioid peptides ha macology of pioid eptides , cadem ic u blishe , H , in press .

- G . B . J . L . B . B oodman , C , Cadet , Em ilien , and R . . Rothman Down regu lation of m u opioid bi nd ing ICV N PFF : t . N I A r M ono r . sites following continuous infusion of and mo ph in e an autoradiographic s udy D Res . g , in press .

G . B . E J . . B I oodma n , C B mil ien , L Cadet , and R . . Rothma n Chron ic CV infusion of morph in e and PFF - l N down regu ate m u opioid receptors in rat brain : a quantitative autoradi ographic study . Reg ulatory

P . eptides , in press

PUBLICATI O NS

J S P B K . . . B . Xu,H . . a rt i ll e , . R . d e Costa , C Rice , and R Rothman Differential bind i ng of opioid peptides and oth er d rugs to two su btypes of opioid d ncx binding sites i n mouse brain : fu rt her evid ence for (1 - 907 . receptor H eterogeneity . P eptides ,

N J S Pa r til l a P . A . S k F . I . r . A . B r . B . S X u . . i , Q H . , , ta r , Ca roll , G i ne , and R Roth man tereochemical i ever s ible 3 - Meth lfe nta n l requirem ents for pse udo rr inhi bition of opioid mu receptor binding by the y y congeners ,

- - 06 3 . : . S R T l 461 44 and its enantiomers evidence for different binding domains ynapse ,

- C . N S Pa r til l a B . . K . . B . S v u . . i , Q H . X , J , R de Costa , Rice , and R Roth man electi e la beli ng of ka ppa ZA opioid recepto rs in rat brain by [1 25I ] I O XY : interaction of opioid peptides and other d mgs with m ultiple - 1 293 . . P ka ppa ZA bi nd ing sites eptides ,

- . . Xu X ...... B J Fl A B . Y A J K M . . ertha , C . , i ppen nderson , R Roth man , H , Cha , E acobson , a nd C Rice - - l N I A M no 5 a lmor h a ns . . o r . l ndolo ry p as potentia selective opioid receptor probes D Res g ,

Xu X . . S G . Y B K W J B ilsk F . r A ...... P N . . Calderon , . , C eorge , H , Cha , R Rothma n , D ild , E y , o reca , E J K c S s acobson , a nd . C . Ri e ynthesis and a bsolute configuration of optically pure ena ntiomer of S N CBO - non e idi d . N I A . BW373U 86 . Developm ent of , a potent and selective p pt c opioid receptor agon ist D Res

o r . Mon g ,

- i H . . . u C . K K . B . P Y . . X Cha , X . , H , i m , C R ce , and R Rothman robing the opioid receptor com plex with

- - w . N I A u erfit: . M ono r . trans s p resolution of t o dcx bi nding sites D Res g ,

ka a a N J S Pa rtill a . . K . B . B Xu . . . . Z i , Q . , H . , , R de Costa , C Rice , and R Roth man Resol uti on of pp 1 251 I O XY . N I A . M ono r opioid receptor su btypes in rat brain using [ ] D Res g . ,

J S P l e . . K . . B l rt B . . Xu , H . , . . a i l , R de Costa , C Rice , and R Roth man Differentia bi nd ing of opioid peptides and other dru gs to two su btypes of opioid d ncx binding sites in mouse brain : fu rt her evid ence for d Mono r N I A . receptor heterogeneity . D Res g . ,

ma n ' a nd L J E J k R . . R oth F . P T oli odeox n u cl eotide . B ils B . ai , y , _ , orreca reatment with antisense g y to selectivel inhibits dZ- a - 5 ntinocice io N e 1 0 2 . t n . eu r oR ort the opioid d receptor y . agonist p p ,

Pa rtill a . . . . N J S . K B S fi . u . B A . rs s B X . H o zta . i , Q . , H . , , R de Costa , C Rice , o odi , , and R Roth man - - -3- - cl ocinna m ox Dihydrocodei none hydrazon e , di hydrocodeinone oxime , naloxone ome oxim e , and fa il to N e u r och e k . m . . irreversi bly in h i bit opioid appa receptor bind ing Res . , i n press

Xu B . . . : . : . N B I Roth ma n , R , Q i , a nd H uprenorph ine a review of th e bind i ng literatu re n B : U O J W W A W A . L uprenorphin e Com batti ng Drug buse ith a nique pioid , Cowan and . . ewis (eds) , J ohn iley

I N Y . S ons , nc . , ew ork , in press

- . B ilsk S . . . B . F P J . F W L S E J . N A . u . . M cN tt H . Xu . Calderon , , R Roth man , orreca , lippen nderson , R . , , E m ith , y , -4 P K . . P S . Davis , a nd C Rice robes for narcotic receptor mediated phenomena . 1 91 . ynth esis of a R -4-a lI l - -1 - i er a zin I -3- m ethox benz I - N N -dieth l benza mide S N C [( ) y d imethyl p p y ) y y ] , y ( a h ighly d e d elta O on e ti . J M n . selective , p p pioid receptor agon ist ed Chem . , i n press .

P . N . B n S . N . K . . E J r F . . B B ilsk or eca , , R er stein , Calderon , C Rice , R . Rothman , a nd . . y A S N C80 - N I A e M ono r i ntinociceptive profile of , a h ighly selective , non peptid ic d elta opioid agonist . D R s . g n press .

- Xu ...... Y . . S N K F P B T Cha , X , H , Calderon , C Rice , orreca, and R . . Rothma n h e non peptid e delta _ i BW373 U86 rts i agon st , , enant omers , and related compounds: i nteractions at multi ple dcx bi nd ing sites in rat

M ono r . . N A . brain . I D Res g , in press

Xu . til l a . B . J . S . a r K S kiri H . . Ka a . y , , R Roth m an , H , P , and C Rice ynthesis and biolog ical eval uation of 6a - - -1 7- -4 5a -e o 6 I O O i l x mor i N I A . b h na ns . M n r . and D d hydroxy methy , p y p D Res . o og , in press SUMMARY OF WOR K

P P P rs m t ri l l hencyclidine ( C ) and sigma recepto , thoug h linked i n ter s of their h is o ca deve opment , and their "Rfl e t w - a r ha r ma colo ica ll a nd generally high affi nity e c ions ith opiates , e now known to be p g y

. T P P M biochemically distinct bi nding sites he C binding site , beca use it is but one component of the N DA o d m N M A recept r, is relevant to g abuse research because the D receptor plays im porta nt roles in neu ronal v processes invol ed in su bstance a buse , such as memory , propagation of seizures a nd ki ndl ing , tolerance

. P P P 3H T CP P P and dependence revious studies showed that the C a nalog , [ ] la bels two h ig h affi nity C “ binding sites : PCP site 1 (N M DA - receptor— associated) and PC P site 2 (biogenic-ami n e -tra nspo rt er

- - . F I v I 4793 1 4 associated) Collaborative investigations with . y Ca rroll lead to th e discovery of RT , which has f r P P r l e high potency and selectivity o C site 2 . Th is novel py o has a n eurochem ical profil e consistent with that

. A P of an antidepressant patent application on this compou nd was filed . M ore recent C P site 2 ligands show - promise as potential anti craving medications . PU BLI CATIONS

B B S W H . Xu . B . . B . S C r F . l . . . . . M a sca r el l a a rol l , , E lough , , , C G oodman , and R Roth m an ynthesis and

- - - - - 1 b r r ol es . P P h exa h d r o Z l m eth l ind e no Z M . . ligand binding at C sites 1 and 2 for y substituted y [ , jpy ed Chem

- 1 82 . Res . ,

l . . . . . M a sca r el l a B N . P B . J . L . F . B B S W Good man , C . D . Thomas , A ert , Emilien , Cadet , Carrol l , E l oug h , , RT l -4793- 1 4 M ki S S . B . A o a ws . . . R g , u brama n ia m , and R Rothman , a new liga nd with h ig h affinity and selectivity for the - M K- 801 -i nsensitive [3H] 1 -thienyl) cycl oh exyl] pipe ri di ne bi ndi ng site (PC P S ite 2) of

- 65 . . S guinea pig brai n ynapse ,

T h u r ka uf . . . . . A M . K M L A A . . F P C . . . . . r A kun ne , H . , J onn , , E Jacobson , C Rice , J T inders , Q J iang , or eca , and B A - M K801 P P 1 P P R . . Roth ma n n electrophilic affi n ity ligand based on distinguishes C site from C 9 -3 . 1 89 . N eur och em . site 2 . Res ,

l . wski . . E B G F . r B M A S S B 8 . . . R o a . B . . . milien , , C oodman , Ca roll , lough , g , u bramaniam , a nd R Roth man - 2 4793 1 4: t P P . N I A . M ono r TI . R a new ligand with high affinity and sel ec ivity for C site D Res g ,

J C . J . P . r S . F S l . . . r e . B W . M a sca r ell a . B DiCesa , , u g ess , , Carroll , and R Roth ma n ynthesis a nd S H -benzoc l - - J r a l c ohe ten S 8 . . structu determ ination of y p , i mines Heterocyclic - 1 92 . Chem . ,

- . B . P P S 2 MK801 . 30. Roth man , R C ite : a hig h affinity i nsensitive phencycl idin e bi nding site - N eur otoxicol . T er a tol . , 353 .

- h J . B . G . B . . E . M J S P J L J W G Uhl B . . . rt e ...... oodma n , C , m il ien , C D ersch , a ill , Cadet , D Vandenberg , a ng , R , b F l B . B K . P . ine rr . B . enza . . Ca ol l , loug h , Constable , and R Rothman Discovery of a n ovel ch iral zep R T I -4793-41 ena ntiosel ectivit P 2 derivative , , whose enantiomers bind potently a nd with moderate y to PC site

l A . N I A M ono r . . and c oned D transporters D Res . g , i n press

- - M . . . Xu Y . . . B . M M J . . F A h . X K B K ert ha , C , V attson , L l ippen nderson , R . B . R ot man , H , Cha , ecketts , and . C Rice A m ark ed change of receptor affinity of the 2- methyl -5 -hyd r oxyphenyl)mor pha ns u pon -8- E m v 5 . attach ment of a n benzyl idene oiety: the synthesis and e aluation of a new class of receptor ligands J .

M . . ed . Chem , in press

- M . . . Y . . B . M G . B H u K B k K t . . r X X . e ha , C , C attson , C eorge , R . Rothman , . , C ha , ec etts , and C Rice I ntroduction of a n - 8- benzyliden e moiety in the Z- methyl -S - ph enylmorp h a n system a bolishes opioid effects and N I A Mono r . affords a n ew class of potent sig ma receptor ligands . D Res . g . , in press SUMMARY OF WOR K

N oninvasive brain imaging is used to study biological corr elates of d rug a buse in hu ma n vol unteers . Positron emission tomography (P ET) and electroencephalography (EEG) show that euphor i ants reduce

"

CM R lc r CM R I c . global and reg ional cerebral meta bolic rates for glucose ( g , g ) , and i ncrease a power Changes in rC M R glc ind uced by morphine or coca ine and in EEG a power produced by morphi ne are

- correlated with positive affect . Cocaine induced changes in rC MR g l c but not in b power are related to

. B a onist/a nta onist CMR Ic rC M R I c mood uprenorphine , a mixed opioid g g , decreases regional g ( g ) , as do I n other a bused d rugs , such as morphine , cocaine , and nicoti ne . su bjects with histories of cocai ne a buse - 3 ongoing stud ies suggest that cocaine related stimuli produce d rug craving , arousal (desynch ronization of rC M R lc P activity) , and i ncreased g in areas of the lim bic system . atients with a cqu ired i m mu nodeficiency A I S syndrome ( D ) show metabolic defects in the lim bic system , consistent with an involvement of the d A I S hippocampus a nd relate areas in cognitive deficits associated with D dementia . Drug a busers that are seronegative for h uma n immunodeficiency virus manifest lower r C M R gI c i n. visual association co rt ex and higher rC MR gIc i n orbitofrontal co rtex than non -d rug a busi ng controls matched for age and socioeconom ic

. . M R lc status To determi ne wh ether these differences predate or result from d rug a buse , rC g and reg ional ' blood ow w t M fl . cerebral _ _ are studied i n t ins discordan for drug a buse agnetic resonan ce imaging is used to r study brain st ucture , as related to personality and cognition in su bstance abusers . Vol u metric ana lyses O reveal no evidence of ventriculomegaly in su bstance abusers who are othe rw ise healthy . ngoi ng stud ies u r assess potential differences in str cture of the orbitofrontal co tex , a brai n region that shows a difference in MR l c rC g i n d rug a busers as compared to controls . Studies i n newborn sheep assessed the effects of O cocain e on cerebral blood flow ( EF) . A sing le dose of cocaine almost dou bled CB F at 30 5 whil e arterial O EF 5 blood pressure was increased ; returned to control withi n min . I n sheep that received cocai ne C B F 30 f repeatedly , s a ter the fifth dose was stim ulated as after the first , and remained el evated for 1 h . w - Cerebral vasodilation , com bined ith hypertension , may contri bute to coca ine associated n eonatal neurologic damage . An i mproved form of a model for calcu lation of (CMR gI c) from PET data has been developed to obviate th e need for art eria l blood samples or any blood samples collected early after i njection of the rad iotracer. PUBLI CATIONS

M MJ Ca scel la N G S J M P R L Y B CK W DF S EK . L . S v or gan , , tapleton , hilli ps , ung , ong , haya ondon ED ensiti ity : i to su bjective effects of coca in e in d rug abusers Relation to cerebral ventr cular size , A m J Psychiatry - 1 7 1 7 .

I n : L S . ED ondon . eeing the brain : The role of P ET , autoradiography , and other i maging tech niques

B A H t . iomedical pproaches to I ll icit Drug Demand Redu ction , ar el CR , ed P roceedings of the I nternational - - 4 03 3698 2 1 . : N I A N I . Research Conference , G eorgia D , H pu blication no ,

'

P A M EJ L T r a stm a n R J . O B r ien T , G leason C , J ones D J r, Cone , ondon ED , y Cerebral responses to si ngle and - a r Pedia tr 343 . multiple coc i ne injections in newbo n sheep , Res

L E P N M ono r . u I A ondon D ositron emission tomography in studies of dr g a buse , D Res g Z4 .

W S L G S F J S J M P R L Da nna ls F S J P KL G alsh , ilson , asinski DR , tapleton , hillips , R , ch midt , reston , rayson R , B G E S JT Contor e i L E B igelow , u lliva n , gg C , ondon D . upren or ph ine reduces cerebral gl ucose m eta bol ism i n rs N eu r o s ch o h - u a nn a colo 70. polydr g a buse , p y p gy 1

P R L E . M l hillips , Hem ing R , London D orphine effects on th e spontaneous electroencepha logra m in po ydrug - h lo -1 8 1 : N eu r o s c o ha rrn a co . abusers Correlations with subjective self reports , p y p gy

M S J L E . I A . tapleton ondon D mag ing techn iq ues , Encycl Drugs lcohol , i n press

I G B J K 8 P R L L - EG Hem ing R , lover , oeppl , h il lips , ondon ED . Coca in e i nduced increases i n E al pha and beta

t e u r o s ch o h a rrn a col . ac ivity evidence for reduced cortical processi ng , N p y p ogy , i n press

E . P n L . ondon D atter s of nicoti ne action in the brain , Tobacco Control , i n press

ri J N B P M B W C S J M L . A ewlin D , reto us , ong , tapleton , ondon ED cute intravenous coca ine educes cardiac N I A M on o r . vagal ton e i n cocai ne a busers , D Res g , in press

P R L Y W DF L n A hillips , Chen C , ong , o don ED . n improved m ethod to calcu late meta boli c rates for gl ucose

P ET J N ucl M . using , ed , i n press SUMMARY OF WORK

I n vitro , ani mal and h u man studies are perform ed to elucidate mechan isms of opioid action a nd to develop t new modalities for treati ng opioid addic ion . Chron ic mo r phin e treatment prod uces dependence a nd tolerance as demonstrated in electrophysiological assays of synaptic activity in the isol a ted spina l cord from neonatal rats . Chemically and electr i cally evoked responses of nociceptive n eurons show opioid tolerance ' b co- . B that is not prevented y treatment with d izocilpine ased upon studies in slices of rat brain , that show

‘ inh ibition of nitric oxid e synthase (N O S ) dim inishes the development of tolerance to m orphi ne in r cem l eus L - no adrenerg ic neurons of the locus ( C) of adult morphine treated rats , new studies i n intact anesthetized rats investiga te the role of LC as a cellular su bstrate for the in volvement of nitric oxide in opioid “ f withdrawal . N ew behavioral experim ents demonstrate that various N OS inh ibitors consistently reduce r a t ' a nd ‘ certain withd rawal sig ns in the , yield pharmacological profiles similar to that of clonid ine . 7 N itr oinda zol e 7 -N l N OS N OS ( ) , a in hibitor specific for cerebral , atten uates more withdra wal sig ns than other

- N OS . F rt 7 N I k v O N O inhi bitors u h er , lac s the asopressor activity common to ther S inh i bitors , suggesting that 7-N I O k holds prom ise as a possible treatment for opioid . addiction in h u mans . ther wor i n rats shows that behavioral signs of opioid withdrawal and widespread stimu lation of cerebral g l u cose m eta bolism occu r when “ meth l na loxoniu m n r L - y , a q uater a y opioid antagonist , is injected directly into C of morph ine depend ent rats . The find ings dem onstrate that increased metabol ic activity i n regions of the brain when withdrawal is i precipitated by per pheral opioid antagonist ad min istration is most likely due at least in. part to sti m ulation of - L . I n u i the C d r g naive rats , buprenorphine m mics the effect of morph i ne on cel l fi ri ng , but i n morph ine r dependent rats bupreno phine mi mics naloxone , consistent with the hypothesis that the actions of rt A buprenorphine , a pa ial opioid agonist , vary with the state of opioid dependence . n assessm ent in h uman volunteers of the interactions of the calciu m channel antagonist verapamil with morph in e shows that verapam il antagonizes th e positive affective changes produced by morphine and potentiates th e analgesic effects of morp h ine . PU BLI CATIONS

K AS B L . N r i A imes , Va upel D , ondon ED it ic oxide in op oid withdrawal : ttenuation of som e withdrawal sig ns by r B -524 inhibitors of nit i c oxide synthase , Psychopharmacology ( erl) .

- B L WR L . Vaupel D , ang e , ondon ED Effects of verapamil on morphin e ind uced su bjective effects analgesia J - and respiratory depression i n humans , P harmacol Exp Ther 1 993 ;267 : 1 386 1 394 .

B L WR L . r r Vaupel D , ange , ondon ED Verapamil potentiates mo phine analgesia and red uces eupho i a i n human

N I A M ono r . subjects , D Res g , in press

L K AS B . I ondon ED , imes , Va upel D nhi bitors of nitric oxide synthase and the opioid withd rawa l syndrome ,

N I A ono r . D Res M g , in press

- B K AS L E . ri 7 nitr oind a e i Vaupel D , imes , ondon D Compa son of zol with other nitr c oxide synthase in h i bitors as Ps chO h a mi a colo atten uators of opioid withdrawal , y p gy , in press .

PU BLI CATIONS

- rt : S u T P Delineating biochemical and functional prope ies of 5 receptors Emergi ng concept . CRC - 3 N 20 . C ritica l Reviews in eu robiology ,

W A J M c JF L P S u A eissma n D , D c ann , orden and T n absence of changes in sigma receptor su btypes d 33 t . . . P . 2 . in the brains of genetically dystonic ( ) rats Eur J harm acol ,

- J S rv r I n : S S u T P and JL u nien igma receptors in the central ne ous system and the pe i phery . The igma -4 2 1 4 . Y z k . A P L Receptors , Ed . lt ha , pp , cadem ic ress , ondon

J A W T -P S u S 1 z : M cCa nn D , D eissman a nd igma and sigma sites in rat brain Comparison of

- . S 1 89 . regional , ontogenetic , and su bcell ular patterns ynapse ,

S N S lzenwa sser B H ell er JS G utkin M M - N N P . S u d T . A Calderon , , , , , attson , and H ewma n ovel 1

' box lic a cid de v a lka neca r ri a tives 1 . J . M l c cl o . . hen . p y y y are potent and selective sig ma ligands ed Chem , i n press

M T T -P S u DW P ri A P ri P E-084 5 P P r - aurice , , a sh and vat R , a selective C de ivative atten uates M K 801 I ea min in m . P m B B . induced impair ent of g m ice har acol . iochem . ehav . , i n press SUMMARY O F 'WO R K

T l l l here a re large individ ual differences among humans and ani mals in behaviora , physio ogica and toxicolog ica l responses to drugs of abuse . These ind ividual d ifferences in behaviora l responses to dru gs

' also display su bstantial genetic and environmental influences , are continuously distributed r a nd appear to “be two For influenced by many g enes rather than one or major genes . th ese reasons , appl ication of several of the techn iq ues of geneti cs and molecular biology could be helpful in identification of genetic i nflu ences in - ex er im e nta l readily stud ied p an imals that could shed light on hu man interindividual differences , and identify genes potentially responsi ble for behavioral variation in drug responses in these a nima ls .

' Differences between several inbred mouse strains nor ma l responses to psychomotor sti m ulants and " " kind li ng ind uced by repetitive d mg admi nistration have been soug ht and cor related with strai n -to -strai n i GA BA A rt d fferences in biochem ical parameters related to opiate receptor , receptor, a nd dopam ine transpo er / - densities and or function . Recom binant in bred strains have been tested for cocaine i nd uced locomotor

- responses a nd sensitization . An imals from a heterogenous stock have been tested for cocai ne ind uced locomotor activation , a nd have been used to esta blish a genetic selection parad igm to ask if selection for

- - cocai ne i nduced locom otion also selects for other cocaine ind uced functions , incl uding psychom otor stimu lant conditioned place preference measures . Each of these approaches wil l allow im proved assessment of genetic i nfluences in su bstance a buse , and analyses of the recom binant in bred strai n comparisons will al low chromosomal mapping of specific genes contri buting to the strai n differences throug h quantitative trait locus a pproaches .

R PHS 6040 ( ev. PUBLICATIONS

JO M R J . i Miner L L , Elmer G I , P ieper , arley Aggress on mod ulates genetic influences on mo rp hi n e a nalgesia as

— 2 . assessed using a classical M endelian cross analysis , Psychopharm acology 2

M J S K Fr i M S . v arl ey R , h imosato , eman , Goldberg R Time course for the d e elopment a nd persistence of the b B anticonvu lsant effects of ca r amazepine against cocaine seizures in three strains of mice , ra in Res .

- 200.

M A I S S K B k McC lea r n P k R W S a rl ey R J , Coll ins C , El mer G , udakov , el nap J , G E , ic ens , G old berg R . G enetic ru I n approaches to u nderstand ing the actions of d gs of abuse . : P roblems of d ru g dependence 1 992: Proceed i ngs of the s4th An n ual S cientific M eeting of the Col lege on P roblems of Dru g Dependence 1 992; N I DA

- Res Mong r 5 1 .

J S M l R S erd ikoff S L A A J S . annerud CA , ar ey , , lastra G , Cohen C , Goldberg R Tolerance to th e behavioral : P m a J P effects of chlordiazepoxid e har acological and biochemi c l selectivity , harmacol Exp Th er 1 994;267 : 1 3 1 1

20.

M J S K E G I M L arl ey R , h imosato , l mer , iner L . P harmacogenetic approaches to d rug dependence . I n : - S L GG . B t L P P W . : onnacott , u nt , eds ioch em is ry of Drug Dependence ondon ortland ress , 1 72 , 1 993 .

t R J - G w M M r T E G l S S . GABA e iss , a ey , horndike B , o d berg R , chindler CW receptor l in ked ch loride channels and P l the behaviora l effects of naltrexon e i n rats , harmaco B iochem B ehav 1 994; in press .

- M iner L t ; M arl ey R J . C hromosom al mapping of loci influencing sensitivity to coca ine i nd uced seizures i n BXD Ps ch O a 9 4 in Recom bi nant I n bred stra ins of mice , y ph rm a coI 1 9 ; press .

- K . r R J . Shimosato , S aito T , M a ley G enotype specific blockad e of cocaine induced weig ht loss by th e protein S 1 994 in . synthesis i n h i bitor, a nisomycin , Life ci ; press SUMMARY OF WORK

t The goal of this projec is to elucidate the nature of the dopami ne transporter, a key cocaine receptor , in term s of its function and as a protein . The character ization of this protei n as a cocaine bind ing site and as an entity of the dopami nerg ic neuron is essential i n understandi ng its fu nction . O ne of the key findings a bout the dopamine transport er is that there appear to be different post translationally modified forms , at least with regard to its mol ecular weight . The d ifferences in molecu lar rt r weig ht appear to be due at least in pa to changes in glycosylation , a lthough it is not possi ble to u le out we l l n . I n v n other sources of heterogen eity as a rece t study , we ha e found that over developme t and aging ,

'

rt . A a n i the m olecular weight of the dopam ine transpo er changes gain , a significant pa rt of th is ch g e s due to differences in glycosylation . ,Th ese results i ndicate that if dopam inergic neu rons are used from i m mature rt l l v species , for example for transplant , th e transpo er is not the same as that i n the fu y de eloped adu lt . T B he mechan isms that regu late the dopami ne transporter are not ful ly unde rstood . ecause of the new w l uta ma ter ic r kno ledge that g g synapses result in the elaboration of nitric oxide , and beca use dopam in e g ic l uta ma te r ic ri w synapses have been fou nd adjacent to g g synapses in st atal tissue , e tested whether or not d . W nitric oxide would in hi bit opamine uptake e found that nitric oxide inhi bits dopamin e uptake in a ti me , dose and tem perature dependent fashion . The in hi bition of dopam ine upta ke occurs at lower concentrations r i k of nit c oxid e generator than does the inhi bition of gl utamate u pta e . These are the first demonstrations that nitric oxide affects neurotransmitter uptake and could have profou nd im plications in the way dopami ne

. oa l transport is reg ulated The next g is to show that this regulation occurs in vivo . P rimary cell cultures of em bryonic rat mid-brai n cells can be used to study the transpo rt er with certa in

. S cl ea r cells w advantages ince it is that these are not withi n the brai n , we wanted to test hether or not the r r dopam ine transporter in these p i ma y cultures was simi lar to that in mature rat brain at least , i n rega rd to their pharm acology . We fou nd that cocaine analogs inhibited dopam ine u ptake in these cu ltures i n a fashion l that did not differ from that in adu t rat brai n . We a lso showed that the solu bilized dopam in e transpor ter can T will read ily be studied by standard receptor binding techniques . h is facil itate a variety of experim ents . W to l e continue make su bstantia progress in understanding the nature of the dopam in e transporter protein , how it is processed and possi ble mechanisms for its regulation . PU BLI CATIONS

B A Pel a r a t B JW Fl Via l M K MJ R osten e W . P A I rouard , p D , oja , Carrol l , , uhar , otent cocaine nalogs , n hibit _ [3H] Dopa m ine U pta ke in Rat M esencephal ic Cells in P rima ry C ultures: P harmacological S electivity of 75 1 3 - 1 7 1 993 S . . B Em bryon ic Cocain e ites Dev rain Research , , ,

T -5 N eu r or e i 1 25 R l 5 B rt . or t 5 1 57 Patel , A . Character zation of [ ] inding to Dog Caudate Dopamin e Transpo er p

1 60, 1 993 .

A . P R . K P . u M . J . O A . f atel , , Cerr ti , C Va ughan , , and uhar , Developm entally Regu lated G lycosylation

. . B Dopa m in e Transporter Dev rain Research , in press .

H J . B M K M . N O I U k B S . r 3H . 1 Rogun , , au man n , and uhar, it i c xide n hi bits [ ] Dopam i ne pta e rain Research 64 - 4 83 91 , 1 99 .

- . M S . K .J . i O 3 H G S R ogun , , Dawson , V and uhar , N itr c xide I nhibits lutamate Transport i n ynaptosomes .

S . ynapse , in press SUMMARY OF WOR K

The purpose of this project is to continue to assess the neurological status of individ uals who have a bused dm s i coca ine and other g over a long per od of their lives . The acute n europsychiatri c disor d ers include h H w seizure , psychosis , su barachnoid hemorr age , and throm boem bolic phenomena . o ever, there are few studies which used the detailed classical neurological exam ination in order to classify possi ble different W v neurolog ical syndromes that might be associated with the use of drugs of a buse . e ha e thus started to carry out thorough neurological and neuropsychological exam ination i n su bjects who are chron ic coca ine or r I polyd rug a buse s and who are seronegative for H V . “

The results of these exam inations continue to show evidence of nystagmus , a bnormal eye pu rsuit , sa cca d-es abnormal , decreased reflexes , a nd increased jaw jerk . Vi bration and position senses were also a bnormal . Th e presence of nystagmus and increased ja w je r k i n these su bjects may be related to cocaine effects on brainstem pathways . Th e reflex and senso ry abnormal ities appear to correspond to a

' ri bilateral sym met c neuropathy . These r esu lts suggest that cocaine may cause deleterious effects on th e v r v u ner ous system by causi ng const i ction of th e vasa ner or m which supply the per i pheral n e rvous system . These findings suggest a new line of investigation which wi ll focus on the per ipheral effects of this wel l known vasoconstri ctor . PUBLI CATIONS

L e n in n N J . L W . R H n fie ld . a d R E . Cadet Gendron , T ange , g . J Rothma n . Th e Clin ica l eurolog ical ,

i i i A : P F . N I A M S i 1 41 : 3 1 1 Exam nat on n C hron ic Cocaine busers relim ina ry indings D onograph er es 994 .

PUBLI CATIONS

- A l l . R . S P . S L . a nd J . A Cadet , , heng , Roth man , Carlson Epstein C . tten uation of metham pheta m in e i nduced co er/zinc m - N e u r och e . 62 380 3 3 . . : 8 1 994 neurotoxicity in pp superoxide dism utase transgen ic m ice J .

- h e im B . l . L . L a d en J . B C uZ . n C u ZnS Cadet , , au m , Carlson and Epstein C superoxide dism utase ( O D) transgen ic mice show resistance to the lethal effects of m ethyl e nd ioxya mph eta mine (M DA) and of B 1 994 methylenedioxymetham phetamin e rai n Res . ( ) in press . SUMMARY OF WORK

These studies conti n ue to assess the distr i bution of [1 25I1R T l -55-I a beled dopami ne a nd serotoni n upta ke

- . o of A sites i n the rat brain Coca ine exerts its addictive properties throug h the bl ckade D u ptake sites . S r i n tudies have been don e to characte ize the specific effects of cocai ne on the brain , usi ng l ga ds wh ich bind

at more than the DA uptake sites . Effort is being made to develop ligand that are more specific to DA

- . O e 1 25I R T l 55 W i uptake sites ne of th se ligands is [ ] . e have determined the specific d istri but on of these I n 6 sites i n th e rat central nervous system in a num ber of studies . addition , we evaluated the effects of A O H D lesions in the n igrostriatal DA pathway . U n ilateral lesions of the nig rostriatal dopam inerg ic pathway 6- O A - 6-O H DA k were performed by the stereotaxic application of H B i n the caudate putam en . caused mar ed tota l 1 25 I I b rt decreases in [ ] RT inding sites in the N A c and the C Pu . Fu her analyses indicated that these e r 5 - HT k d creases cor esponded to changes in both and DA u pta e site s . T hese stud ies a lso h elped to n 1 25 I I -55 5- HT - demo strate that [ 1RT bind to sites in the caudate putamen a nd in the frontal co rt ex of rats . These experiments stress the impo rt ant of both biochemical and lesion studies in order to characterize the

anatom ical d istri bution of monoami nergic uptake sites in the rat brain .

v PHS 6040 ( R e . PUBLI CATI O NS

A . t l Pa rt il a J . S . B . R . L O r d inez S . . M . r B k B k K . M . J . . K . C Cadet , , , C De sch , roc ington , , ec et s , , . , de Costa , Rice .

- k nne . B A utor a a h ic R . F . I A u H Q dio r O H Carroll , , , and Rothman , Quantitative g p Evaluation of th e Effect of 6 l - L B S L A 1 25 l I 55 . N I A M S r Dopam in e esions on inding ite abeled with the Cocaine nalog , [ ] RT D onograph e i es

1 41 : 252 , 1 994 .

J . L . G . U e J . S . . . R . K . C . P I n e H . C . M P B . A kku n , Dersch C , Cadet Char artill , de Costa , Rice , Carroll and rt I l l . B . S . B 5 Rothman R . tudies of the biogenic am ine transpo ers Demonstration of two bindi ng sites for G R 1 293

- . P . 268 1 462 1 47 994 3H B T C P . Ex . . : 5 1 and [ ] in rat caudate mem branes J harmacol p Ther ,

k L . A u nne H . C S il er thom . J . v L B . M . e J . S . . . R . M B P B R K . C Rothman , Cadet , auman n , artil l de Costa , Rice ,

S . J U l a J . h Pa rtil . B . l . . l . W G J S Carroll P . , , ang , G lowa and Dersch C M . tudies of the biogenic a m in e transpo rters . I V . Demonstration of a mu ltipl icity of binding sites in rat caudate for the cocai ne a nalog [1 25 I1RT I PET 55 . J (i n press) SUMMARY OF WOR K

Th is project assesses the safety and efficacy of pha rm acological treatm ents of cocai ne and opioid a buse/d ependence in clin ical tr ia ls . Evidence from preclinical studies suggests that the rei nforci ng effects of coca ine are related to its i nhi bition of dopamine reuptake . M uch of the work to develop pha rm acol ogical treatments of cocaine dependence has th us far focused on dopam inergic agents; though no dopam in erg ic

A . agents have yet been shown to be effective i n red ucing cocaine use . n open tr ia l testing th e safety and r r efficacy of com bination treatment with buproprion and bromoc i ptine , agents with dopa mi ne g ic activity , is nN a l ow u nde y and has so far shown a incidence of side effects among treated su bjects . This study is the first to apply the strategy of com bining pharmacologic agents to increase the efficacy of ind ividual agents in the treatment of cocaine dependence . Data collected to date suggest that the com bination of bupropion a nd bromocripti ne is safe and possi bly effective for treatment of cocai ne dependence

T a S he pa rt ial opiate agon ist buprenorphine is a s fe and effective treatment for opiate dependence . ome preclin ical stud ies and uncontrolled clinical case seri es have suggested that buprenorph i ne mig ht also be - w . A r effective in reducing coca i ne use by opiate addicts double blind , control led cl inical trial is u nde ay that directly eva luates the effi cacy of buprenorph ine in r educing both opiate and cocaine use in dually opiate and

- cocaine dependent patients .

M edically su pervised withdrawal from opioids is a common ly used treatment but is usual ly not effective in establish ing long -term a bsti nence b ecause patients frequently relapse soon after com pletion of the withdrawal . A procedure for initiating naltrexone maintenance during withdrawal treatm ent is being developed to provide a more effective post-withdrawal treatment that includes pharmacological treatment ' bu with opioid a ntagonists . The effica cy of prenor phine/na ltr exone combi nations is being tested in a clinica l rs the l tri al . The fi t phase of study has been completed . The results suggest that administration of na trexone on day 2 may i ncreases O piate withdrawal at the beg inning of treatment but l eads to decreased withdrawa l thereafter, com pared to treatment with naltrexone placebo . This r esult may bear on the hypothesis that rt antagonists may produce pa ial resetting of the opiate receptor, resulting in a shorter duration of withdrawal . These data also support the development of cost-effective short-term inpatient opiate detoxification a nd - earl y transition to opiate free treatment using naltrexone . PUBLI CATIONS

L . T P a . d l K S v . . B . B t a one P n . . reston , , ul li an , J , erger, , igelow , G E Effec s of cocaine and in com bination wrt h n e r ime nta l mazindol i n hu ma n cocai ne a busers . Jour al of P harmacology and Exp Th erapeuti cs 1 993 ; 267 2 96 7 30 .

i bson A . C . . L . L e . . . S . P K B tra in E , reston , , , I , and igelow , G E P recipitated withdrawal by pentazocine in

- methadone mai ntained vol u ntee rs . Journal of P harmacology and Experi mental Thera peuti cs 1 993 ;

634.

S C K . L . S . L . L ie bso B A . . . . . P M n I . train , E , reston , , titzer, , , , and igelow , G E The effects of cocai ne i n - : buprenorph i ne maintain ed outpatient volu nteers Resu lts from cli nical experience a nd la boratory challenges . A J n t 1 994 - The me r ican our al of Addic ions ; 1 43 .

W S . L . P K . L . S M . L . B E . J . . alsh , , reston , , titzer, , Cone , and igelow , G E Cli nical pharmacology of f - : . P m 1 994 buprenorph in e Ceiling ef ects at h ig h doses Clinical har acology and Therapeutics ; 580.

P K . L . J . . S M . J k P tr a n . . . sna sa l reston , , T ull iva n , Testa , , and D R asins i sychopharmacology of butorp hanol . A c 1 994 - Drug and l ohol Depend ence ; 1 67 .

L . F . . . R . S G S J . S J P Da nna ls S P W M . . . L . . E alsh , , ilson , , asinski , D , tapleton , , hill ips , R , , R , ch m idt, J . , reston ,

R . . E. T or e C K . G B S . L B L . Cont i . . . rayson , , igelow , G , ullivan J , gg , , and ondon , E D uprenorphin e reduces r N - r . 994 cerebral glucose meta bolism in polyd ug abuse s europsychopharmacology 1 ; 1 70. SUMMARY OF WORK

- The section has continued to investigate two major non ph a nr ra col ogica l treatm ents of su bstance a buse .

‘ v Last year a project was completed showing that a beha ioral intervention , contingency management , was P effective i n decreasi ng cocaine use among a group of methadone mai ntenance patients . atients i n the contingency management group achieved substantially longer periods of sustained a bstin ence from cocaine - use than persons in a yoked control group . A follow up study is unde rw ay to refine th e rei nforcem ent protocol to i mprove rates of initiation and maintenance of abstinence . Th e original rei nforcem ent schedule was designed to escalate in value so that the incentive grew as the length of ti me that patients were

. th e r e wa s initia l v abstinent grew Thus , a strong incentive to remain abstinent . The va lu e of th e ouchers , v n v i I howe er, was small , and a um ber of su bjects never ach ie ed in tia l a bstinence . n th e current study , a bon us was added for th e fi rst few cocaine- negative ur ines and then faded out i n order increase th e proportion of su bjects achieving abstinence; the escalating incentive val ue was retained to maintai n absti nence .

The contingency management procedure is also being appl ied in a second study to i m prove compliance with na ltrexone treatment . Cli n ical experience has shown that compliance with naltrexone treatm ent tends to be i cl inica l T fi very poor, ser ously com promising its utility . he purpose of the study is to test the ef cacy of contingency management for im proving patient compliance with naltrexone treatment .

Counseli ng is an import ant element of vi rt ually a ll drug abuse treatment; thus standardization and eval uation of cou nselin sol e _ g procedures is needed to establish effective treatment , whether counsel ing is the treatment h a nn a cother a - or is given i n com bination with p py . A 1 2 week study was com pleted i n wh ich th e effi ca cy of a standa rd ized individ ua l Cognitive/B eha vior a I/l nter persona l counseling program adm i nistered accor d ing to a k a . specified therapy man ual , given either twice weekly , once wee ly , or every two weeks , were comp red O - rt utcome measures i ncl uded cocaine and other drug use (by self repo and urine toxicol ogy) , coca i ne P craving , psychological state , and psychosocial functioning . rel imi na ry a nalysis i ndicated that there were - w k fewer in itial drop outs in the t ice a wee treatment group , but no significant differences in d rug use outcome measures between grou ps after twelve weeks of treatment. The manual is currently bei ng adapted for use in a group treatment format . ' PUB L ICA T I O JN ‘S

PUBLI CATI O NS

L . . . K . G . K k B . P K . S A S G 8 . M m J . . M . K . umford , , Evans , , a i ns i , , reston , , annerud , C ilverm an , and riffiths R

i a . R . Discr mi native sti m ul us and su bjective effects of theobromine c ffei ne in hu ma ns

- Psychopharm acology 1 994; 8 . SUMMARY OF WO R K

The psycholog ica l assessment of su bjects pa rt icipating i n cl ini cal trials has become an im porta nt part of the ' section 5 research interest . Severa l studies were conducted : a comparison of d rug a busers see ki ng treatment and not seeking treatment; an evaluation of personality disorders i n cocai ne depend ent individuals rv seeking research treatm ent; and a su ey of motivation for treatment . An im po rt ant issue faci ng research ers and treatment providers is the repo rt ing and representation of women S 6 1 and m i norities i n cocain e a buse pharm acotherapy clinical tri a ls . ociodemographic representation in r En lish reports of outpatient pha macotherapy for cocaine abuse , pu bl ished in refereed , g la ng uag e journals between 1 983 a nd 1 993 was com pared to the epidemiology of frequent cocaine use i n th e 1 990 N I DA " - National Household S urvey of Drug Abuse (N I DA H S ) . The fi ndings showed that i mportant t rt rt sociodemog raph ic data is of en not repo ed in papers , and raise issues of quality of research repo s , l general izabi lity of resu lts , socia eq u ity , and accessi bility for certain groups in cocain e a buse pha r m acotherapy research .

I n proced ural studies to id entify im proved outcom e variable in cl i nical trials , th e usefuln ess of q ua ntitative

' urinalysis for cocaine meta bolite and creatinine co r rection tech n iq ues and the relationsh ip between these data and self-reported d rug use were assessed with data collected in a clinical tria l (N 37) of a contingency a management beh viora l treatment i ntervention . Rules were d eveloped to differentiate between occasions of rr - P new use and ca y over in positive q ualitative urine tests . reliminary analyses suggest : q ualitative and quantitative u ri ne testing show greater rates of dru g use tha n that shown by self report; quantitative testi ng provides a means of differentiating incidences of new drug use from residual carry -over; the identification of w u ne use with q uantitative testing may help to reconcile differences between rates of dr g use indica ted by . - qua l itative urine screens and se lf report .

Sweat patches have been proven effective in measuring coca ine and opiate use in controlled cl in ical la boratory settings but have not previously been tested under more realistic settings such as our treatment r cl inic . Coll ection of sweat as a method of mon ito i ng drug use may be more accurate a nd rel ia ble over a l ri i r T onger pe od of time than the ur ne testing method cur ently used . he purpose to eval uate the t n effec iveness of collecti ng sweat as an alter ative method of monitor ing drug use . Results wi ll be com pared u w to dr g use as mon itored by uri ne samples collected three ti mes a eek . PUBLICATIO NS

P D P . J . R E : s ch o a t o . M v v S l M I J . F A h o ontoya , H ess , C o i . udala , ; Joh nson Comparati e tudy of y o gy and O r Cognitive Functions between Cocaine and piate Dependent P atients Seeking Treatm ent . A me ica n Jou rn al of 1 994 -42 Addictions ; .

- C . A . M D . H a ertzen P n N I . A I P ontoya , ; , Reduction of sychopathology mong ndivid uals a rticipati g i n on treatment

- - S . n Drug Abuse Residential tudies J our al of Addictive Diseases 1 994; 1 3 : 89 97 . SUMMARY O FWOR K

Sweat was evaluated as a media for use in monitori ng drug use by h uma n su bjects . H ea lthy su bjects with a t histo ry of chem ical su bs ance a buse volunteered for these studies . I nform ed consent was o btai ned and all I B F procedures were approved by the hospital nstitutional Revi ew oard . ollowi ng the ad mi nistration of O k cocai ne , marijuana or opiates , sweat samples were collected periodically . ther biological specim ens li e i a nd a lso S saliva , blood , ur ne hair were collected . pecimens were analyzed by im mun oassay a nd gas a to r a h /ma ss chrom g p y spectrometry .

e in These data wil l provide n ew_ information on this u nusual biological specimen wh ich may b useful

' of m ethod s development for monitoring human d r ug exposure . PU BLI CATIONS

. M . Da J . R rw rn W H v M . k A K S F H D . Cone , E . J i l lsgro e , , Jen ins , eenan , and . . weat Testi ng or eroin , Cocaine

A A . 1 994 . nd M eta bolites . J . nal . Toxicol in press , SUMMARY OF WORK

T v t l he organ preser ation effec of opioids is exami ned in a sing le organ preservation preparation . Rat ungs l A E A L are dissected and preserved in hypotherm ic solutions with or without a de ta opioid peptid e D D L . D D E h oth enn ic T t markedly enhances the yp preservation ti me of isolated rat lu ngs . h e fu nc ions of th e hypother rn ica lly prese rved lu ngs with added DADLE are compara ble to the functions of th e controls which

e . L rv A LE are d issected a nd stud i d immediately ungs prese ed with out D D functioned poorly i n the test . PUBLI CATIONS

‘ S P O elt en J N R K A a nd T - P S u x rv v m u lti r n Chien , R g , Diana , lley E tension of tissu e su i al ti me i n o ga

C a r d iova sc . . 967 block preparation using a delta O pioid DAD LE . J . Thorac . S urg

PUBLI CATIONS

- H K M S M N J L L . F 3 ife n d il ashi moto , antion e C R , pada R , eumeyer . ondon ED urther characterizati on of [H 1 pr o

Eu r J 77 . binding in rat brai n , P harm acol

K L I S S Y Hashimoto , ondon ED . maging igma receptors and cerebral responses to igma drugs . I n : ltzh ak ed . - 4 s . : 2 2 . The S ig ma Receptor M iami Academic Press , SUMMARY OF WOR K

The paral lel li ne bioassay is a classical statistica l technique used in pharmacology to estimate the relative com O Und U t l r d ni . r potency of a test p relative to a standa d g nfo unately , this comp ex a nalysis is not avai la ble mmer cia l t co w . r on statistica l analysis soft are Therefore , sho cut methods , that a re not based on a valid v - . A a statistical process are used to obtain relati e potency estimates user friend ly statistical a nalysis progr m , B I O AS SAY entitled , using the para llel l ine bioassay being as core proced ure , is being d eveloped a nd written ++ in C for use with D O S systems . The program is designed to make more efficient use of balanced ri experimental desig ns , such as those used for human and p mate pharmacolog ical stud ies , but it may be we l A lso used with in vitro studies as l . incl uded are other statistical tests which are useful in analyzi ng dose ' : v S - B response data linear regression , Dunnett s test , missing al ues calcu lations , cheffe ox test for i t- rm OTU S homogeneity of var ance , tests and a data transfo ation routine . Data may be im ported from L

files , and there are three ways to output the final analyses . The uniq ue statistical adva ntages of the program : v include the calculation of relati e potency estimates and confidence l imits based on li near, parallel portions of two dose-response cu rves for equi - effective d rug effects; partitioning out a Between S u bjects va riance w l ' component ith the para lel line , li near regression and Du nnett s tests; calculating missing values for random ized block experimental designs and making the appropriate corrections in the A N OVA ; and having w - i i . e the capability of making inverse predictions ith confidence limits from l near dose response curves ( . d m estimating the dose of a g requ ired to produce a given effect) . Th e scope of the project encom passes l A progra m deve opment , validation studies and preparation of a n instruction manual . t th is tim e , programm ing is complete , validation is in progress , and a d raft of th e instruction man ual has been prepared . PUBL I CATIONS SUMMARY OF WORK

With rega rd to their analgesic effects , opioids have been thought to act on receptors withi n th e centr a l

- rv . ne ous system excl usively Recently , however, we have shown that , within inflamed rat paws , i m mu ne cell

‘ der ived opio id peptides can activate peripheral opioid receptors loca ted on sensory nerves a nd ind uce r a ntinociception . The pu pose of our current research is to examine agents for their capa bil ity of releasi ng S opioid peptides from im mu ne cells and for their potential i n the inhi bition of pain . uch periph eral effects are t w O of considera ble interes in vie of the avoidance of centrally mediated side effects of pioid analgesics , such ri O u r as dyspho a , dependence , addiction , sedation and respiratory depression . most recent experiments have exami ned whether co rt icotropin releasing factor (GR F) or interleu ki n - l B eta (I L -1 B eta) release opioid a d r esult - peptides in inflamed tissue n in analgesia . U pon administration of GR F or l L 1 B eta into both paws ’ hind a w of rats with un ilateral p inflam mation , nociceptive thresholds increase markedly in the i nfla med but

- - - noninfla med . A GR F 1 not i n the paw lpha helical and interleukin receptor antagonist , respectively , t a GR F l L - 1 antagon ize this analgesic effec , indic ting that and Beta act via their specific receptors . These receptors are most l ikely loca lized on immu ne cells within the inflamed tissue because i mm u nosu ppression by cyclospo r in A atten uates the effect . I n experiments with antisera against opioid peptides we have shown cells T that endogen ous opioids released from i mmune mediate these analgesic effects . h is is suppo rted by r the fact that naloxone and other opioid antagonists everse th ese effects . These results suggest that O R F I L -1 B w and eta , by activation of their receptors on i mmune cells , cause a release of opioids hich r rv O subsequently occupy their receptors on senso y ne es resu lting in in hi bition of pain . ngoing in vitro experiments exam ine whether CR F or l L -1 B eta are capable of releasi ng endorphin i n cell suspensions prepared from inflamed and noninfia med lymph nodes and whether this release can be atten uated by the rt " respective antagon ists . To fu her elucidate the mechanisms governing the apparent u pr eg u l a tion of O pioid rv i n receptors on sensory ne es dur ng i flammation , we are examining the permeability of the peri neurial barri er by histochemistry and the expression of opioid receptor genes in dorsal root ganglia . PUBLI CATI O NS

- 1 3 rt S M rt L S . I k chaefer , Ca er , tei n C nterleu i n and co icotropi n releasing factor in hi bit pain by releasing opioids

- N a tl S U S 23 . from i mm u ne cells in inflamed tissue , P roc Acad ci A

A nton r evic M SA S M S . P j l , ousa , chaefer tein C eri neurial defect and peripheral opioid a nalgesia i n 4 J N 1 99 . inflam mation , euroscience ; in press

Cz onkowski E l e - S S M L l A M S n . t k a ntinocice tion tein C , chaefer , Carter , , ousa , pp C Cy o in e induced p mediated cel ls 1 9 - by opioids released from i mm une , Regulatory P eptides 94;5 0: S l 91 2 .

M SA M WM H A H S L S t ousa , itch ell , assan , Carter , tein C . Cor icotropin releasing factor receptors in inflamed P 1 994 . tissue : autoradiographic identification , Reg ulatory eptides ; in press

- - - S M rt L S . l l rt chaefer , Ca er , tei n C l nter eu kin and co icotropi n releasing factor i nduced rel ease of IS endorphin cel ls P 994: r 1 . from im mu ne and inh i bition of inflam mato y pain , Regu latory eptides in press SUMMARY OF WORK

This project assesses the efficacy and safety of new pharmacologic and bio logic treatm ents for dru g ex erimenta l l S ome a nima l dependence using p parad igms in a controlled , residentia envi ronment . and open i- v label hu man studies suggest that the ant convulsant carbamazepine may reduce coca i n e cra i ng and use , l - e possi b y by blocki ng the development of cocaine i nduced kindling . However, a recently com pleted dou bl blind study did not support the efficacy of carbamazepin e i n the treatm ent of coca in e d ependence .

I w k n hu mans , very lo calorie d iets producing etonemia are associated with the absence of hu ng er, but it is k a lso I a not n own whether this su bjective effect applies to d rugs of a buse . n animals , bala nced low c lorie m - diets not prod ucing keton e ia increase drug self administration . A second com ponent of th is project is k eval uating which of these d ietary effects operates in human d rug a busers , using nicoti ne (cigarette smo ing) “ th e r . P r as ta get drug rel imina y results indicate that a balanced low calorie diet i ncreases cigarette smoking , l i a lter . whi e a low calor e ketogenic diet does not smoking These fi ndings have clin ical i m plications , especially since some drugs of abuse themselves suppress appetite a nd th us may prod uce cal orie depfiva fion .

ri r The p mary enzyme meta bolizing cocaine in h umans is buty y lcholinesterase . I n theory , alterations in ' enzym e activity m ight alter brai n levels of cocaine and its m eta bol ites and th us alter coca ine s effects , with I n possi ble therapeutic benefits . a colla borative study with the Preclinica l Pharmacology Laboratory and the a lt National I nstitute on Aging , com pounds which er butyrylcholinesterase activity are g iven to m on keys to r w dete m i ne hether they alter th e acute effects of cocaine . PUBLI CATI O NS

k M s no m a l M L J F . P r Crowell D , Ches in , usial revalence of gastrointestinal ymptoms in obese and weig ht

- i J Ga str oenter ol 391 1 994 . binge eaters . Amer can ournal of ,

sca te l l os N ikoo b L k L J Mo SA ma nesh P . S a sesses ampo rt RD , Ches in , terile epidural and bi lateral psoas in a ' - e . n r e A Ga st o nter ol 1 089 1 994 . patient with Crohn s diseas merican Jour al of , ,

. I n : i k B k L B Z v . r L J . P P A Ches i n Gastrointesti nal bleed ing ar er R , urton J , ie e D (eds) nciples of mbu lato ry M W W k B 4th . 1 994 . ed ici ne , ed i lliams and il ens , altimore

PU BLI CATIONS SUMMARY O F WORK

i n l Drug a buse h as ser ous , ofte letha , biomedi cal consequences , such as H IV i nfection and ca rd i ovascular

- . t ri t c k rs dysfu nction Th is project studies su bject charac e s i s , h ig h ris behaviors , and other facto associated ' h e O with such consequences , with th e goal of developing interventions to prevent or a melior a te t m . ne study rw v - currently unde ay provides a com prehensi e , non invasive assessment of cardiovascu lar function i n 24- r EKG rt cocaine abusers , using hour a m bu lato y monitoring of , blood pressure , and h ea rate ; high KG r resolution E ; analysis of heart rate va ia bil ity (vagal tone) , and echocardiography . Eval uation of the subcl inical effects of acute and chronic cocaine use on card iovascu lar function wil l help elucidate th e ' v mechanisms of cocai ne s cardiovascu lar effects , hopefully leading to their pre ention or am elioration . PU BL I CATIO NS

PUBLI CATI O NS

K K M W L k A rw WD EJ : ato , H illsgrove , ein hold , Gorelic D , Da in , Cone Cocaine and m eta bolite excretion in i i nonstiu ml a ted -341 . n A sal va u nder st mulated and conditions Jour al of nalytical Toxicology , , 1 993 .

k M 8. k . r k . S tein Ja rv i E , G orelic DA I mpairment of memo y by fluoxetine in smo ers Experimental and 1 : 1 88- 1 93 1 993 Clinica l Psychopharm acology . SUMMARY O F -WORK

Th is project is eval uati ng factors associated with drug users seeking treatment in a hospital em ergency

. u ri c l room Dr g users may present to a hospital emerg ency room for a va ety of reasons , som e cl i ni al y - appropriate a l ife threaten ing med ical consequence of dru g use) and some less effici ent. uses of the - T health care system non urgent medical condition , attempt to enter su bstance a buse treatment) . here is little data available on the relationship between emergency room visits by drug users and th eir seeking of rt and pa icipation in drug abuse treatment . This study , in colla boration with the Depa rt m ent of Em ergency M k B M B M o edicine at the Hop i ns ayview edical Center, altimore , D , collects data on the sociodem graphic , - drug use , and treatment seeki ng behavior of an unselected series of d rug users visiti ng a n urba n hospital emergency room .

Another com ponent in colla boration with the Clinical Dependency U n it at the H opkins B ayview M ed ical

Center, is evaluating the infl uence of medical morbidity a nd prior medical and su bsta nce a buse treatment rt- on the response to sho term inpatient detoxification treatment for su bstance abuse . PU BLI CATIONS

i B J C . W H S u r ! all , . hy as It P roved o Difficult to Match Dr g A buse P atients To Approp ate Treatment - 6 4 2 6 1 99 . Addiction , ,

PUBLICATIONS

J H S J r l l . . . o e H ennin fie d . E . g . , eish man (in press) The addictive of n icotine i n tobacco

Psychopharm acology .

M . . . . m m l S L . S P t ennin fie d . H g , J E chu h , , Heishman , J (in press) har acological deter i na nts of cigaret e

k P B 8 r k M . k F . X . A d l kofe r K . Ed s t smo ing . I n . . . Cla e , Qui , , Th urau ( ) , Effec s of nicotin e on biological i h a u ser Ve ri a 000 B a se l : B r k . systems (pp . g

H S e nn in field . E . . I ! J . H eishman , . , g , J (i n press) s caffeine a drug of dependence Criteri a and

Pha rm a co s choecolo ia . comparisons . p y g SUMMARY OF WORK

T dm s t w he mechan isms by which g affect human behavior are complex , involving the interac ion bet een th e ' d irect actions of the d rug im paired coord ination) and the fu nctiona l beha vior a l effects such as altered m . S motivation pecific d eter i nants of drug response include the drug dose , the route of ad min istration , the ' rt person s physiological a nd psychological state , the pa icular environ mental conditions , and the natu re of the O ne behavior or. test used . approach we are pu r suing is to vary the environ mental conditions by in u l a tin k T ma p g the reinforcement (monetary) contingencies u nder which su bjects perform various tas s . h is ' r can be thought of as ma nipu lating a subject s motivation to perform . By also va y ing drug dose and using tests that m easure different aspects of performance psychomotor vs . cognitive) , we ca n beg in to u explore the complex interactions underlying the effects of d r gs on behavior .

Another approach to investigating the mechanisms by which drugs influence behavior is to focus on

- performa nce i mpairment produced by psychoactive d rugs . The per fo r m ance impairing effects of a bused d rugs produce a large tol l on the nation each year in terms of traffic injuries and fatal ities and lost

t r . w k produc ivity in the wo kplace However, for most d r ugs , e lack basic nowledge a bout the behaviora l m mechanis s u nderlyi ng their impairment of human per formance . A series of stud ies is being conducted that

will . A r address such questions battey of physiolog ical , behavioral , and performancemeasu res desig ned to determi ne whether an ind ivid ua l is behaviorally impaired as the r esu lt of taking a d rug has been tested with

. A S 1 994 f ethanol , marijuana , and cocaine study scheduled to begin in eptem ber, will exam i ne the ef ects of amphetam ine , codeine , and alprazolam on this same test battery as wel l as a cognitive test of a ttention and lso . a i memory By collecting blood samples dur ng these studies , we will be a ble to gai n valua ble inform ation l r concern ing the re ationsh ip between plasma concentrations of d ugs and degree of performance i mpairment . PUBLICATIONS

H S J L : v eishman , . . (i n press) . aboratory performance assessment l mpairi ng effects of psychoacti e drugs .

N A M W U S v n O . I D Research onograph . ash ington , DC : . Go er m ent P rinti ng ffice

C . H en nin field S . J . . . . . N k : A v Heish ma n , , Taylor, R , g , J E (in press) icotine and smo ing re iew of effects on

m . x P human perfor ance E perimental and Clin ical sychopha rm acology .

H S J . S k m eishma n , . . (i n press) trengths and wea nesses in the appl ication of laboratory perfor ance assessment l . A xe . . . I S . Ed s M W k S N I A to wor kplace settings n H D J Crouch ( ) , Research ethods i n or place ettings . D

M . W : U S . P Research onograph ashington , DC Govern ment rinti ng O ffice .

Y . . J . H e . in fie ld F M . S F S nn J f k k iore , , hi , , Heishman , , g , . E . The ef ect of smo ing and sm o ing withdrawal on 994 m : A 1 . v flight perfor ance u pdate Report to Centers for Disease Control and P re ention .

S .J F . R . H en S nin field . J P E. m Heish man , nyder, and g . erfor ance , su bjective , and physiolog ica l effects of - - o . u A 34: 1 1 n ic tine in non smokers Dr gs and lcohol Dependence , 1 8 , 1 993 .

PUBLI CATIONS

J . H e nn i n fie ld E. P k W B . B k . C . . E B Cohen , , ic worth , , un er , and g . Caffeine antagonizes EEG effects of n icotine 7 9 - P r e mi r B 4 : 1 9 926 1 994 . withdrawal . harm acology B och st y a nd ehavior .

H e nnin field J . E W . . P rt B . P I n : Cohen , C ickwo h , and g , harmacologic characteristics of tobacco dependence . 545 - P H et a l 1 993 . 5 revention of Respiratory Disease , irsch (eds) , pp 58 .

W . B . . I E . P k rt P n : N n eld J . H e nn i gfi , Cohen , C and ic wo h , sychopharmacology of n icoti ne icotin e 9 - l . O 1 93 45 . O S 4 Dependence . C . T r ea ns , J lade (eds) , xford P ress , pp . 2 .

R M . H en nin fi d e B . el J . E . : M lch a nis k W . K N m I k P ic wo rt h , , eena n , and g , icotine Effects and s . n : Hand boo of toxicol o 2 M k N Y 1 994 N eu r o gy , Volu me Chang and Dyer, (eds) arcel Dek er, ew ork , (in press) .

B . E B . H en nin . ld E . m P rt W B fie J . ickwo h , , un ker, and g , Transd er al nicoti ne ; red uction of smoking with mi ni mal - . P 9 1 4 1 9 4 . abuse l ia bility sychopharm acology , 9

D e M . F . B . H nnin fi eld J . P S W . B . sch k E. r B ut y , , ailey , , g , and ickworth , moking without nicoti ne delive y d ecreases 1 -h r t w 2 . P B B 1 994 ithdrawal in abs i nent smokers ha rm acology iochem istry and ehavior, (i n press) . “ A ew a p u n st sa rpnts a sa uxur or j srsA reue exec) s r oter oq enoo J no li q a utuo u r 1sa i | ertrugu e ua A tfi ua a q seu ' U O I S JS A a sa uruo 8 l s exa l u r ua uuM se a ben fiu e| usrueds pr ep u ets a ur u r a teJetu seM pue | a A a | O u tpeer ep eJ O utu a A a s 8 mean re p eu us e; a a e a ou etsq nskrod j o M otsru p eu oda J e uwvr sa xel ‘ uotsnoH ur O Jj sunp a to p a sod ur oo seM ejdtu es a ur s emis patru n out ur sunpa 6u r>| ea ds usru eds pue ater a m0 1 a ou eA a ra J s uo e su er s ssa sse o a en eA a bu a s UO S JS A s ued o ° uo en eA a n a , u | t tu1 i p t | t q t l u r s ur n | Burofir a pu a r e ei ep u; pue

‘ oe n o ua a a A e sa n s M eu ur a i fi u l a u u a 1.)t pa to puo q u tp t r u d r t g r sr otefirrsa A ur utrM uotter oqenoo ut pa d0 | a A a p ‘ seM UO lS J S A a sa uruo e pue euma or ea ur sr oi efirtsa /tur li q pa dora A a p seM U0 !S J S A usrueds v t ua ur nnsut ‘ a m utrM p a tdur a ue ua a q seu mom | eJn1| no-ssor o emu tnq M otua A U| oa v out st jertua tod a A rtorpp e Jra ut a zua toer euo p ue a a e to sfimp to swa y a | eor60 | oq sd a ut a teutuenb oi tua ur n ns u r pasn A | a prM 1s0 tu

MB OM : l o A B VWWfl S

A b uu e eo u qoun omoe ucj | r r| o

uouer g morooe u ueucj | B O l Ut| 0

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PU BLICATI O NS

E . J . O f C . Y . H . G . A . M H T . M A , W . D oldberger, B . A . , Da rw in , Grant , , llen , , Caplan , and Con e easurement eroin - - 3 9 670- 67 5 99 . : 1 3 . And Its M eta bolites B y I sotope Dil ution Electron I mpact Mass S pectrometry . Clin Chem . ,

B . A . w W . D . k T . M r k . P B .A Cone , E . J . , Holic y , , G rant , Da in , and Gold berger, , ha rm aco ineti cs and " - 3 l 1 7 : 327 3 7 1 993 . A na . . S . J . P harm acodynam ics of l ntranasal norted Heroin Toxicol , ,

W P S z M . E . J . B G . P r B : K . alsh , S . , reston , , tit er, , Cone , and igelow , , Clinical ha m acology of u prenorphine

1 993 . H . . P . Ceiling Effects at igh Doses Cli n harm acol Ther. , I n P ress ,

S . Pa u l 8 O F F . U M . u L D . M J . Cone , E . J Dickerson , , and itchell , , orensic Dr g Testi ng or piates V ri ne Testing F M A O J 1 7 : - W . . A . . 1 56 1 64 or H eroin , orp hi ne nd Codeine ith Com mercial piate l m munoassays nal Toxicol .

1 993 .

A 6-A cet l r B . m J G E . J . E GC/M S A o h ine P A . J . . . . enkins , , old berger, Cone , and Hoey , D ssay of y p i n lasma - P M S A N M S 9 - Hewlett ackard pplication ote , 3 3 .

- G B A . E. J . Confi r ma tor F u W old ber ger, and Cone , y Tests or Dr gs in the orkplace by gas ch romatogra phy mass J Ch oma to r I n P r 1 994. spectrometry . . g ress , .

R O . B . rw E J . u S M s . F T L L r . S . L . W . D . t mith , . . , Hughe , , evi ne , , Dicke son , , Da in , and Cone , orensic Dr g es i ng

F O VI . U ri T t F O O W . A or piates n e es ing or Hydromorphone , Hydrocodone , xymorp hone nd xycodone ith I O G Chr oma to r a h /M a s S ctr ometr . J . A na l . . P Commercial piate l mmu noassays and as g p y s pe y Toxicol , n ress ,

1 994.

B . . G l B . A L w rw W D . E . J . I ntr a su b ect i O S o dberg er, o enthal , , Da in , and Cone , j Var ation f Creati nin e , pecific I n I n 9 4 . G A H M U i S U . . P ravity nd p easurements Consecutive r ne pecim ens in Heroin sers Cli n Ch em , ress , 1 9

E . P w rt W . B . B W P . E I B P S A T ick o h , , un ker, , elch , and Cone , ntravenous u prenorp hin e Reduces u pil ize nd he L I n S 49: 1 -1 3 9 ight Reflex H u mans , Life ciences , 29 8 , 1 90. SUMMARY OF WORK

u . When d r gs of abuse are smoked , volatile components and pyrolysis material escape into the atmosphere

Depend ing on the loca l environ ment , bystanders may be exposed to the dru g by passive i nhalati on of the contam inated ai r.

Artificial methods have been developed to smoke d rugs of a buse in a controlled environ ment a nd to measure dru g air levels . These methods are then applied to the desig n of human clinica l studi es to assess the hazards of environ mental exposur e to drugs . S ix subjects were exposed to vaporized cocaine in a small unventi lated room Th e study was perform ed under blind conditions with placebo control and two active

. U r o t doses of cocain e i ne , bl od and saliva samples indicated that su bjects were exposed o cocai ne , but sufficient amounts were not absorbed to produce pharmacological effects or test positive at standard cutoff concentrations .

U nknowin dr u ex osu r e g g p could be dangerous to unsuspecting bystanders , pa rticularly to sma ll chi ld ren . i wrll i These stud es esta bl sh limits of exposure to volatile comp onents of drugs under controlled conditions . PU BLI CATI O NS

' - A . O B r ie n J r . . . . T r a stm a n . C . H J . P . M . F Gleason , . , l ida , T , . Douglas Jones , E Cone and R J y etal Responses

- n S . 34 H 9 H 1 4 1 993 . A M a ter na l I I . A . P To cute Cocaine njection heep mer J hysiol , ,

'

A . J r . D . E . J . E . a st a n . J . P G C . J L r m O B r ien , T . . leason , , , ones , , Cone , , ondon . D and T y , R Cerebral S M I n N S t i 3 5 - A I n . P Responses To ingle nd u ltiple Coca ine njections ewbor heep edia r c Research , ; 339 343 4 1 99 .

PU BLI CATIONS

C M L . S err n ine i rt R itzM , antion e CR , ondon ED p interacts with cocaine bind ing sites on dopa m ne transpo ers , 4 4z4 - Psychopharm acology 1 99 ; 1 1 7 52 .

F S U L J I n v J E F J . H lesher , cheffel , ondon ED , rost ivo la beling of nicotin ic cholinergic receptors i n brai n with [

- L S 1 994 54z1 883 1 890 . 3]cytisine , ife ci ;

I L S K B L N L N ngram E , l ijima , uo H , resnaha n E , G reig H , ondon ED . ew pharmacologica l - NY S strategi es for cognitive enhancement using a rat model of age related memory impairment , Ann Acad ci

- 32 .

- - - L A . P N t D NY ondon ED , M ukh in olyami nes as endogenous modulators of th e me hyl aspa rt ate receptor An n

A S . ca d ci , i n press

I DK S L S i K G N L ngram , pangler E l ijima , l kar H , uo H , reig H , ondon ED . Rodent models of memo r y A : M L S dysfu nction in lzheimer s disease and normal aging oving beyond the chol inergic hypothesis , ife ci in press .

‘ S A S E L L E I n r a m S errn idine - himada , pangler , ondon D , g DK . p potentiates dizocilpine i nd uced i mpairment of I 1 4- T - E J P m ea m ing performance by rats i n a unit maze , ur har acol , in press .

B JA L B e la n L : P N M DA - ell , C g , and ED ondon aradoxical effect of ascorbic acid on i nd uced neurotoxicity ,

R e ort . N eur o p , in press SUMMARY O F WORK

e i T Drug residues have been d tected in human hair specimens by a var ety of a nalytical tech niques . hese rt r i repo s have generated su bstantial interest in using hair as a histo cal record of dru g usage . C u rrent studies rs are designed to determ ine th e presence and time cou e of d rugs of a buse in human hair .

v Healthy mal e ol unteers with a h istory of chemical su bstance a buse pa rt icipated in th e study . I nformed n a ll I B S co sent was obta ined a nd procedures were approved by the hospital nstitutional Review oard . u bjects cl inica l resided on the ward of the ARC . Head and facial hair specimens were obtained prior to and after

. B A administration of d rugs of abuse lood , saliva and urine specimens also were obtai ned . nalyses of tissue biofl u ids m and for d rug was perfor ed by radioimm unoassay and gas ch r oma tog r a phy/ma ss spectrometry .

These studies provide thescientific basis for dete r m ination of the usefulness of hair as a h isto r ical record for su bstance a buse . PUBLI CATIONS

O O f I n O f J W W . H A M H rw . . W Cone , E . Da i n . D and ang , , The ccu rrence Cocaine . eroi n nd eta bol ites air 5 -68 9 63 : 5 1 93 . F S . . rs . Drug Abuse orensic ci I nt ,

J . W . S A H A M I H W . D . E L . rw W . ang , , Da in , and Cone , imultaneous ssay of Cocain e , eroin nd etabolite n air , B S U B - S r ma to 1 99 C h o r . 4 . A r M . . lood , aliva nd i ne y Gas Ch romatography ass pectrometry J g , i n press ,

L . W . H I H A W . Cone , E . J . and ang , ow Environmental Drug Exposure Can mpact air Testi ng for Drugs of buse 4 N M 1 99 . I DA onograph , I n P ress ,

W E. J . H u A I W . L . . ang , and Cone , Testing Human air for dr gs of buse , V Envi ronmental Cocain e Contamination l ntl 9 W F S . P I n 1 9 4. And ashing Effects . orensic ci ress ,

a A . G Da rr . Y . H G B .A . . E. J . O M M M oldberger, , j , , Caplan , and Con e , Detection f ethadone , ethadon e eta bolites , I n I A O I O A O M S . N I A M n P 1 994 . nd ther ll icit Drugs f buse Hair f ethadone Treatment u bjects D onograph , ress ,

Y - A . . H . J G . S O O mor B E. . A 6 A cet l old berg er, , Ca plan , and Cone , ignifica nce f Detection f Heroi n nd y p hine I n I A M 1 994 Hair. N D onograph , in press , .

E. J . t W W . L . T H A A t f O ang , and Cone , es ing Human air for Drugs of buse , V . t em pted Dif erentiation f E a F a U M 1 994 . nvironm ental Coc ine Contam i nation rom Active Coc ine se . onog raph , i n press ,

PU BLI CATIONS

A J C W n LA K . v nthony , a r er , and essler RC Comparati e epidemiology of dependence on tobacco . alcohol , : N S controlled su bstances , and inhalants basic findings from the ational Comorbidity urvey . Ex p e rimenta l and -25 1 994 Clinica l Psychopharmacology . . SUMMARY OF WORK

T will om his study assess th e functioning of g exposed infants with in the .context of their u niq ue environ ments l w u C t ri Ea n ( iving ith a d r g using aretaker) and their own charac e stic abil ities to adapt . y m easu res of the ‘ newborns neur obeha vior a l and emotional adaptability along with maternal and environ m ental factors wi ll be “

related to outcome measu res of physiological , behavioral , emoti onal , and cog nitive functioning during the ' second ha lf of the fi rst year of life . The specific functions targeted incl ude the infants a biliti es to regu late their arousal a nd reactivity to stim ulation , and their development of emotional expressivity , attentiona l r capacity , motivation , and visual recognition memo y (an early predictor of intel lectual fu nctioni ng) .

One h undred d r ug exposed -a nd one hundred non -exposed fu ll -term newborns wil l be recru ited at delivery Ver ification of group classification will be determi ned by 1 ) examinati on of prenatal m ed ical records and 2)

. n 4 9 1 6 1 2 . meconi um analysis Th e infants will be tested as newbor s and agai n at , , , and m onths “ A ne u r obeh a vior a l ssessments wil l include m easures of fu nctioning , autonomic regulation and reactivity to v rt - stimulation ( agal tone , hea rate and cortisol) , emotional regulation during mother infant interactions , r r F T I maste y motivation and attention during st uctured play with mother and exam iner, aga n est of nfant

I B S . M n ntell igence , and ayley Developmental cales ater al assessments include interviews , q uestionnaires , - and observations for d rug h istory and socio economic factors , parenting attitudes , hom e and fam i ly l n v environ ment , psychological intellectua functioning , mater al interactive a nd careta ki ng beha ior .

PHS 6040 (R ev. PU BLICATIO NS

' ' PU E'LI CATI O N S SUMMARY OF WORK

I nvestigators who conduct studies designed to assess fa mil ia l transmission of a vulnerability or resistance to a of g a buse often interpret some their findings as evidence of a genetic determ inant . Fami lies n ot only _ P share genes but they also share an environment . arents i nfl uence their children through modeling and the T establish ment of accepta bl e norms of behavior. h is may i nclude th e form ation of expectations about the effects of d rugs . Th is study is designed to determ ine wheth er instru ctions ca n a lter the mood effects of test

. two I n rt 7 d rugs Th e study consists of experiments . both , pa ici pants are trained to discri minate between 5

. I n h v mg tripelennam ine and placebo the first experi ment , they are told that the capsu les t ey recei e might - k be a sedative , stimulant or placebo but that one of them is more sedative li e than the other . They are also r v i info m ed that their discrimination will occasional ly be tested . mostly with sedati es . These instruct ons are ‘ rt - k designed to bias the su bject to repo sedative li e effects . Th e second experiment is identical except that instmctions r - 4 the are designed to bias the subject to repo t stim ulant l ike effects . During both experi ments , d ru s/doses : 5 - I g will be tested and mg diazepam and 5 and 1 0 mg d amphetamine . t is expected that i v dur ng the fi rst experiment these low doses of diazepam , wh ich previous studies ha e shown do not have - k e L robust sedative li e effects , will be identified as tripelennam in and clearly la beled as a sedative . ikewise , the mal lea ble effects of ampheta mi ne seen in previous stud ies shou ld result in most su bjects identifyi ng at

- 5 k . I r i least th e mg dose as sedative l i e n contrast du i ng the second exper ment , it is more l ikely that a test l drug wil l be identified as placebo and labeled as a stimu ant . To date , 7 participants have com pleted both experiments with an additional two completing the stimulant experi ment only and an addition al one who r completed the sedative expe i ment only . Preliminayr analyses Of the data indicate that discri m ination and drug labeli ng results are consistent with the hypotheses . PU BL’I CATIONS

SUMMARY OF WOR K

The a bility of placebo drug capsu les to serv e as conditioned rei nforcer s as a function of being paired with r differentia l rates of point einforcement was evaluated . N orm al volunteers were ad mi nistered two d ifferently

. A f e . colored capsul es orally in separate sessions lthough di ferent in color, both capsu les wer placebos The

volunteers were told that these capsules might contain a stim ulant , sedatives , or placebo . During each a rt k S session , the volu nteers p icipated in performa nce tas s . u bjective effects and physiolog ical measures

. k were also obtained Th e tas s , which were difficult and ambiguous , were programm ed so that following the k adm inistration of one color capsule , the frequency of reinforcement was mar edly greater than followi ng the

adm inistration of the capsule of the other color . Volunteers were told that the difference i n point earn i ngs - t o h eir . rt was related to th e accuracy t performance During the fi rst experiment , pa icipants were exposed two

w . r f r times each to the t o reinforcement conditions Du ing choice sessions , no per o ma nce tasks were done O rt - but participants were offered the ppo un ity to choose which capsule they preferred to self adm i n ister .

‘ l a r e d iff Duri ng the exposure sessions , th ere were g er ences in th e mood effects associated with th e two i capsul es a nd the reinforcement conditions . Dur ng choice sessions , most pa rt icipants chose the capsul e e associated with the hig h fr q uency of reinforcement . A second experi ment was designed to determ i ne whether the d ifferential mood effects associated with the differential rei nforcement conditions could be A conditioned to the color of the placebo capsu le associated with them . lthough si milar d ifferences in mood i were seen d ur ng sessions when performance testing was don e , there were no differentia l mood effects

observed when the two different colored capsu les were administered in the absence of p erformance testi ng . These results i nd icate that dru gs may function as conditioned reinforcers and thus influence th e proba bil ity of

- d ru g taking behavior.

P HS 6040 (R ev. PUBLICATION S

PU BL ICAT I O NS SUMMARY OF WOR K

This study was desig ned to evaluate the association between fam ily stru cture and th e i nitiation of il l icit d r ug v sa m l e 1 - 7 use a mong a representati e p of 2 1 year old adolescents . The hypothesis was that adolescents who do not l ive with either their mother or father might ar e at an i ncreased risk for starting il licit d r ug use . Th is hypothesis was tested using an incident case-control analyses of self- report data from the 1 992 N ational - Household S u rvey on Drug Abuse . A post stratification proced ure was used to hold constant shared neig h borhood characteristi cs . M ultiple logistic regression models were used to estimate relative risk of ' r of a r e nts initiating illicit d ug use among adolescents residing in homes with va ry ing ki nds p present , holding t 43 1 830 constant age , sex , eth n icity , family income , and num ber of si blings . A ota l of 3 incident cases a nd h - contr ls neigh bor ood matched e were identified . The risk of sta rt ing ill icit drug use was hig hest for fath er mOth er/ste fa ther - moth er/fa ther alone famil ies , followed by p families , no parent families and lowest for a lone a nd fa ther/ste moth er families . M oth er p fam ilies did not show any sig nificantly h igh er risk than mother/fa th er fam ilies . PUBLI CAT IONS

PU BL I CAT I O NS SUMMARY OF WOR K

M uch concern has developed recently over the possi ble increase in hallucinogen use a mong children and es n rs n u . I young ad ults , as some r earche war a bout a possible resu rgence of use of th ese dr gs pa rticu lar, the M r F S rv a onito i ng the uture tudy , using a su ey admi nistered i n schools , fou nd a statistic l ly sign ificant increase 1 991 - 1 992 - - s we ll from in the lifetime and one year prevalence of hallucinogen use a mong eighth g rader , as a l l W as an i ncreasing trend for age groups through young adulthood . e set out to see if si m ilar patterns of halluci nogen use could be obse rv ed using data from three N ational H ousehold S u rveys on D r ug A buse 988 1 9 90 1 99 1 2 . W v G M conducted in , , and e employed Generalized Additi e M odels ( A ) to exam ine th e age specific prevalence of hallucinogen use in th e lifetime and past year . These models use a smoothi ng noni ine ' We tech niqu e to account for l a rity in prevalence across the a ge range wh il e ma ximizing precision .

- ' f_ ound n o evidence of an increasing trend in the one year or lifetime prevalence of hal luci nogens over the last In 1 9 four years . each of the su rveys , year olds were most likely to have used in the last year; however , there is no evidence that th is use contin ues as these cohorts age into their twenties . These results i ndicate that the recent ala rm over 'h a l l u cinogen use should be tempered and that focus on th is class of drugs shou ld not distract attenti on from other drugs . PU BLICATI O NS

SUMMARY OF WORK

Earl y in itiation of d rug use has be shown to be associated with i ncreased risk of later d ru g a buse a nd s e o r development of d r ug dependence . The hypothesis te t d i n this four wave l ngitudinal study of u ban B M r e o schoolch ildren , conducted in altimore D conce ned a suspected causal association betwe n n eig h borho d tota l 925 i deterioration and the ri sk of ea ri y i nitiation of dru g use . A of participants com pleted four confident al I n standardized interviews conducted by specially trained inte rviewers in the spr i ng of four successi ve years . i k addition to d r ug use assessment , separate standard scales were applied to measure a n a rray of r s factors , I ri v B 1 992 c ru . n ludi ng neigh borhood dete oration , peer d g use , parenta l super ision and antisocial behavior y , 340 1 2- 1 4 old rt u k a total of youths , th en years , repo ed d r g involvement (smo ing tobacco , using alcohol

- se ri . A K M without parental perm ission , U of in halants , ma juana , or coca ine) n analysis using aplan eier cu rves showed that th roug hout the fol low- Up interval children report ed earl ier onset of d rug use i n areas rated more

— f. : : rt lo k 2 d . U deteri orated ( g ran test with p value nder the Cox propo ional hazards model , r i a v holding constant other factors (peer d ug use , parent monitor ng , ntisocial beha ior, this association r w h ter til e still pe sisted , resu lting in a hazard ratio of bet een the lowest and ig hest of neigh borhood 95% C l : r r rl deterioration ( Thus , neigh borhood dete io ation appeared to be associated with ea y l a n a nd onset of drug use independent from other suspected factors . We p to extent this i nvestigation use GIS w . geographica l soft are ( ) to include census tract inform ation , wh ich we now have availa ble PU BLI CAT ION S

SUMMARY OF WORK

I n a previous study we demonstrated that inhalant use is a good vulnerabil ity ma rker for i njecting d ru g use .

S A T . T h e study was based on the National Household u rvey on Dru g buse . h e major restriction of our

w w rv - A - - u findings ere that the analyses ere based on a su ey with cross sectional character . s a follow p to that we A A i study , are cu rrently a nalyzing data from a recently completed longitudinal study . coho rt of fr can A i S 82% mer ca n school children have been followed from first grade to tenth grade . ixteen yea r s later of 32 r - T -u those ch ildren , now approximately years of age , have been e inte rv iewed . hese follow p data al low us to test ou r hypothesize that those who used inhalants during their youth are at a greater risk for use of heroin . allowi ng for a clear evaluation of ti me sequence . After elimi nation of those respondents who used h eroi n as i 9 P r adolescents and those with m ssing responses , we were left with 6 8 respondents . reli m i na y ana lyses indicate that after adjustment for sociodemographic factors and adjustment for use of ma rijuana , in hala nt use

' was a sign ificant pr edictor of later h eroin use (RR : 95% C I : I n the S am e model we fou nd use of ma rijuana a lso to be a predictor with a slightly lower point estimate of RR (95% C I of N one of the sociodemographic factors was signifi ca ntly associated with later heroin use . Th ese r l rv findings strengthen our a g ument that ear y inhalant use dese es more attention , not on ly beca use of w lon rt u i k toxicity associated ith g and sho term expos re , but a lso as a vu l nera bi lity mark er for increased r s of injected d r ugs such as heroin . PUBLI CATIO NS

PU BLICATIONS SUMMARY OF WORK

M T ost etiologic studies on drug use , special ly on i njecting drug use , are based on treatm ent sam ples . his study ai med to clarify some potential differences between treated and untreated d rug use rs that mig ht be u considered in studies that only have dr g users recru ited from treatment settings as pa rt icipants . Th is study fu r ther shed light on different suspected determ i nants of entry into a detoxification treatment on ly and entry

- into a methadone maintena nce treatment program . O ur a nalysis were based on baseli ne and six month - u rv u I DU s follow p i nte iews of in itially injecti ng dr g users ( ) , who had been recru ited throug h extensive comm u nity outreach . We first examined characteristics associated with being i n treatment at baselin e and

. A up to on e year before baseline further investigation was focused on active i njecting d rug users , who had ri - not been i n treatment in that time pe od , and who returned for fol low u p assessment with i n 9 months after W m baseli ne . e studied hypothesized deter inants of those who entered a detoxification treatment , and those ri v l who entered a methadone maintenance program , compa ng them to acti e DUs who d id not enter th ese

. O u rt i u tota l treatments f d r g users , who repo ed njecting dr gs at the time of the follow u p i nterview , a of tota l 4 1 44 entered a detoxification program , a of 6 entered a methadone maintenance treatment program . M ultipl e regression analysis indicated that enrol lment in a d et0xifica tion program was associated with a u v r recent episode of dr g o erdose , recent higher freq uency of injecting dru gs , and a h istory of a rest or i . B l on treatment ei ng ma rr ed or living with a pa rtner , female sex , g d uration of drug use yea rs) , and history of treatm ent had separate associations with enrollment into a methadon e maintenance treatment program . PUBLI CATION S

PUBLI CATIONS

I n : . U G e r I L a B ud a M P k R W . u . M HY hl R , El m G , C , ic ens Genetic influences in dr g abuse eltzer , ed 994 P N Y k: P 1 . sychopha rm acology: Th e fou rth generation of progress . ew or Raven ress , : in press

- G I P J O G S G r F . O Elmer , ieper , old berg R , eo ge R pioid operant self adm in istration analg esia , sti mu lation and - m 4 P 1 99 . respirato ry depression i n u deficient mice , sychophar acology ; in press

A r S Elmer I . B mbrosi o E , G old be g R , G ehavior genetic analysis of in nate locomotor activity a nd a cq uisition of

- B v P m 1 994 . morp hin e self ad ministration behavior, eha ioral har acology ; in press

I G S G B G v v A . Elmer G , old berg R , orelick D Rothman R enetic factors i n olved in the acute sensiti ity a nd - P ecifi c 1 994 . context sp sensitization to cocaine , sychopha rmacology ; i n press

S S K G S i S r k L i IV v J G I udakov , old berg R , Bor sova EV , u ova A , Tur na , Rusako D , El mer . Differences i n i re rnfor cement two : rs morp h n e property in in bred rat strains associations with cortical recepto , behavioral sra f P - na l e 8 . activity , a g and the cataleptic e fects of morphine , sychopharm acology SUMMARY OF WOR K

e: I n 1 990 B l rt w DZ Rational , um and associates repo ed an allelic association bet een the dopami ne receptor I ri ' gene and alcoholism . A Taq rest ction fragment length polymorp hism (R FLP) loca ted i n the 3 flanki ng region of the gene was found to be mor e common among alcoholi cs than n onalcoh oli cs [B lum A 1 S et a l . et a l This same allele ( ) has now been associated with other types of su bstance use [ m ith , r DZ 8 1 5 ' wh ile another va iant of the receptor gene ( allele) , located more towards th e region of the B gene , has also been associated with alcohol ism and other drug a buse [ lum ,

ri To furth er cla fy the rol e of these allelic associations in suscepti bility to addiction , param eters wil l be l w D S DZ developed that distingu ish individ ua s ho differ in expression of Z alleles . ince dopa m in e agon ists a I n i B - have been shown to ca use ch nges neuroendocr ne in h u mans [ en J onathan , 1 985] and to i mprove motor functioni ng and a bnorma lities in visual evoked potentials (VEP) in patients with P ark i nson 's disease 1 - et a l 1 99 B W . 1 986 O n fri o et a l . Wachtel , ; odis ollner , ; these will be examined as possi ble i ndicators DZ of d ifferences i n dopam ine allelic status .

T o l v . F h is protoc l wi l ser e two purposes irst , it will allow us to character ize changes in _neuroendocrin e - motor r e ns . response profi le , visual evoked potential , and spo e in non addicted individ uals after ad m i n istration

dO a mine DZ l . S l of the p agonist pergo ide econd , it wil allow us totest the hypothesis that ndivid ua ls DZ w l n i who differ in alleles il have different patterns of responses after pergolide chal lenge . I the r will t protocol , pe g olide be adminis ered orally in low doses to normals without a h istory of d r ug dependence l or recent i l icit drug use , and will not be given in com bination with any other d rug . P'U BL I CA T I 'O N S

PU BLICATIONS SUMMARY OF WOR K

These studies are designed to increase our understanding of the neurochemical mechan isms that are rel evant to coca ine and opioid a buse . Coca ine and opioids such as morphine and heroin are widely a bused ri su bstances that activate dopamine ch regions of the brain , and there are ma ny studies sugg esti ng A involvem ent of opioid systems in the actions of psychomotor stim ulants . natomical studies dem onstrate that d opamin erg ic termina ls are co localized with opioid receptors and opioid peptides i n many brai n t regions . I n an i mal models , cocaine induced reinforcement is at en uated by either d opam ine or opioid v v antagonists , suggesting that both systems are in ol ed in the effects of cocai ne .

The specific objective of this project is to examine whether there are functional chang es i n opioid receptors l -w O in rat brain fol owing treatment ith coca ine , morphi ne , or pioid antagonists . S i nce opioid receptors are l n w negatively coup ed to ade ylyl cyclase (opioids i ll inh ibit cyclase activity) , it is possi ble to measure changes r in the fu nction of opioid recepto s in vitro . Chronic treatment with an opioid receptor a ntagon ist , such as naltrexone , prod uces an i ncrease in the nu mber of mu opioid recepto r s in the brain . Recent stud ies in our laboratory have shown that there are a lso I ncreases i n the fu nction of opioid receptors following ch ron ic

na ltr exone a dministr a tion . W e have also shown that there are i ncreases in these receptors fol lowing chronic , t continuous ad minis ration with cocaine , but that these changes are lim ited to specific brai n regions . F f urthermore , the magnitude of the changes appear to be dif erent following daily i njections of cocai ne (a treatm ent regi men that produced behavioral sensitization) than after conti nuous infusions of cocain e (a l T a r e regim en that produced behaviora tolerance) . hese findings being fu rther investigated . Th ese fi ndings suggest that the opioid system may play an important role in the behavioral effects produced by cocain e and r may serve as a ta get for the development of medica l therapies for cocaine abuse . PUBLICATIONS

E l enwa sser S W H B N u r e u la tion 73 1 Cote T , z , eems , altrexone induced p g of m u opioid receptors on 5c cel l and m brain mem branes : en hancement of opioid effica cy in inhibiting adenylyl cyclase . Journa l of P har acology and T 44 menta l 2 . Experi herapeuti cs ,

S lzenwa sser , . I ncreased m u opioid efficacy for in hi bition of adenylyl cyclase induced by chron ic treatment with

o . 1 994 naltrexon e or c ca ine Regulatory Peptides , ; in press

PUBLICATIONS

- W B Y M P S v k U h l . O r ece tor/bind in : cDN A ang J , l ma i , Eppler C , G regor pi a C . G R m u piate p g protein cloning

- A S 4 . and expression . P N

U : cD N A W J PS P A k AL r A h l . H ang JB , oh nson , ersico , Haw ins , G iffi n C , G R uman m u opiate receptor and ri t F B S L 1 994 338 z - m 21 7 Z . genom ic clones , phar acologic characte zation and chromosomal assignmen E ett ; Z

P S W J B W WF U G . Johnson , ang , ang , hl R Expressed mu receptor couples to adenylate cyclase and N e u r o - r R e ort 9 . phosphatidyl i nositol tu n over, p

- U S P r . A N S 1 994 s I 1 7 z89 93 . hl G R , Ch ilder , aste na k G n opiate receptor gene family reun ion , T ;

J-B M J W J B b M L uth in U h l G L . P i Eppler C , H ulmes D , ang , ohnson , Cor ett , DR , R , inden J ur ficati on and pa rt ial 9 3 268z26447-5 J B iol 1 9 1 . amin o acid sequence of a m u opioid receptor from rat brain , Chem ;

B Xu W J B Pa r til la JS Ka a K G - kiri U h l . Roth ma n R , H , ang , , y H , Rice C , R The ligand selectivity of cloned hu man rs : A 1 2 l ox - t R 1 994 in and rat opioid m u recepto are si milar q uantitative study with [ 51 ] y ago , Reg u ept ; press .

B Xu W JB Pa rt ill a JS Ka a kiri K hl G L - Rothman R , H , ang , , y H , Rice C , and U R . igand selectivities of cloned O rs J P l E er 1 9 h uman and rat pioid m u recepto , harm aco xp T h 94;submitted . SUMMARY OF WORK

M s i r u opiate receptor are the pr ncipal brain sites for the analgesic , eupho i c and addicti ng activiti es of h m o morph ine and heroin . P ar ac log ic data suggest that these receptors contain bi nding sites for agon ists " " l l r and a ntagonists , incuding opiate smal molecu e drugs and la ger neu ropeptides , but deta ils of the means whereby these receptor proteins recognize these ligands are cu rrently unknown .

I n the current FY , mutated , deleted , and chimeric vers ions of the rat and hu ma n mu receptors have been r ece tor/l i a nd i created and studied to refine our u nderstanding of p g i nteractions , and of receptor i ntr nsic - N - - activities and G protei n coupling . terminal involvement i n bi nd ing appeared minima l ; 64 N termi nal a mino a t F 33 acids of the r mu receptor could be removed without effects on radioligand bindi ng . urther deletion of C-terrn ina l resid ues yielded a receptor which recognized mu-selective agon ists morp h in e a nd DAM G O ([D M ePhe4 Gl -ol5 enke h a lin A Ia Z . T , , y ] p ) at wild type levels h ree charged transmem brane residu es of the m u A s -1 1 4 A -1 47 -297 cr itica l receptor, p . sp and H is , were for h igh affi nity agon ist recognition , suggesting ionc t m u/ka a possi ble interac ions with aspects of mu receptor r a dioliga nds . Results of studyi ng chimeric pp receptors in which the putative second extracellu l ar loop of the wi ld type mu receptor has been replaced with that of the kappa receptor fit with a substantial role for this extracelu lar loop in dynor phi n peptide recognition .

These studies su bstantially enhance our working knowledge of means whereby opiate receptors recognize u opiate d r gs and opioid peptides , and provide significa nt clues to new pathways for form ulation of selective w t ne d rugs with ac ivities at these receptors . PUBLI CATIONS

S K P S M i S eid l eck B K B l a scha k CJ W U O A J B . : urr a tt C , Joh nson , or waki , , , ang , hl G R m u piate receptor cri tica l i J charg ed transmem bran e domain ami no acids are for agonist recognition and intr nsic activity , B iol - 53 Chem .

- W J B J P S W J M W WF U h l . k O ang , oh nson , u , ang , G R Human piate receptor second extracell u lar loop ' mu/ka a B 994 elevates dynorp h i n s affin ity for hu man pp ch imeras J iol Chem 1 ; in press .

PUBLI CATI O NS

J PS W JB WF U hl ohnson , ang , Wang , G R . Expressed mu receptor couples to adenylate cyclase and

- n N eu r oR e ort 9 . phosphatidyl inositol tur over , p

J S P A k L r U H : Wang J B , ohnson P , ersico , Haw i ns A G iffin CA , h l G R . uman m u opiate receptor cDN A and

m r F B S L - genom ic clones , phar acologic characte ization and chromosomal assignment , E ett 22 .

W JB J PS I Y P rs AM O zenber er BA M U . cDN A ang , oh nson , mai , e ico , g , Eppler C , hl G R clon ing of a n orp han F B S L ri 9 . opiate receptor gene family mem ber and its splice va ant , E ett

i S ei e S K J PS M r dl ck B K ch a CJ W J E U . O u r ratt C , ohnson , o wa ki A , , B l a s k , ang , hl G R mu piate receptor: i t i v J B charged transmem brane doma in am ino acids are cr tical for agon is recogn ition a nd intr nsic acti ity , iol -53 Chem .

W JB J PS J - M WF U G ang , ohnson , Wu , Wang , hl R . Huma n k opiate receptor second extracell u lar loop '

r mu/ka a J B iol l 9g4 in . elevates dyno phin s affi nity for h u ma n pp chimeras , Chem ; press SUMMARY OF WOR K

T r O he m u , or m orphine prefer i ng , piate receptor is the receptor most identified with th e add ictive a nd i analgesic propert ies of opiate drugs . Prev ous studies with antagonists have identified regu lation of receptor / t F binding properties and or func ion following chronic opiate adm inistration . or these reasons , th e regional r i mu O R mR N A d ist bution and regulated expression of rat m u opiate receptor ( ) and protei n , and reg ulation of this expression , are of interest .

Regional d istribution assessment using a ri bonuclease protection assay revea led that th e most a bu ndant R mR N A mu O . mR N A was found i n thalamus , followed by hypothalam us , m idbrain , a nd spina l cord Th is was

. rt i I ri . also detected in co ex , str atum , brainstem , hippoca mpus , but not i n cerebell u m or pe phera l tissues n situ hybrid ization stud ies revealed that su bpopulations of neurons in several thalamic n uclei express m uO R A R N media l . m , with most a bundant expression i n neurons of the aspect of the lateral ha ben u la

n muo R mR N A I n Developme tal studies d etected i n brains from em bryos as early as 1 4 days gestation . t I n l cerebral cor ex , these levels plateaued after birth . mesencephalon , pons and medul a , h owever, postnatal ' development continued to reach expression levels more tha n 2 fold h igher at 6 weeks th a t a t the end of the

first week of life . Withdrawal from chronic morph ine treatments showed modest effects on rat brai n muO R els mR N A lev that did not reach statistical significa nce .

These studies begin to reveal some of the exquisitely detai led reg ulated expression of th is principal receptor r for rewa d ing and analgesic actions of opiate d rugs . PUBLI CATI O NS

- O e e tor/bind in W J B I Y E M P S k U . r c : cDN A ang , mai , ppl er C , Gregor piva C hl G R m u piate p g protein cloning

- R N S 4 . and expression , A

U h l W J S P rs A L G ri . : cDN ang J B , ohnson P , e ico , Hawkins A , ffin CA , G R H uman m u opiate receptor A and m r F B S L 1 994 338 - genom ic clones , phar acologic characte ization and chromosomal assignment , E ett ; z2 1 7 22 .

U w M JM S I Y W J B U h l K k MJ . M nter ald E , Ru benfeld , pa ngler R , mai , ang , G R and ree u opioid receptor mR N A F 1 9 R e ul 94 in . levels following chronic naltrexone ad mi nistration , g ept ; press

- S M I Y U G S . P i l fi mo chafer , ma i , hl R and tein C er phera analgesic ef cacy of a opioid agon ist i n relation

PU BLI CATIONS

G AM W P A U h l . S rt ex r essron : r onzalez , alther D , azos , G R ynaptic vesicu lar monoamine transp o er p dist i bution - 6 M B 2 . and pharm acologic profi le , ol rai n Res SUMMARY OF WOR K

T e r a nima ls h re are la g e individ ual differences among hu mans and i n behavioral , physi olog ical and c u I ndivid toxico logi al responses to dr gs of abuse . ua l differences I n h uman behavioral responses to d rugs st c appear to display su b antial geneti influences , although these influences may be provid ed by several genes . Fam ily stud ies suggest severa l severe limitations to pedigree- based Iika ge approach es in d ru g a bir se suggesting that association studies might be more fru itful . Use of allelic association a pproach es also ma ndated careful exam ination of ethn ic differences in popu lations and lin kage diseq ui li br i-u m at specific loci that can confou nd these approaches .

Association studi es with polymorph ic markers at several different dopaminergic gene loci ca n test the hypothesis that interindividua l differences in genes of dopam inerg ic neurotra nsmission cou ld contribute to r i ff a inte ndividual di erences in vulnera bi lity to su bst nce abuse . Duri ng this fisca l year, th is la boratory has contin ued to work to develop methods that would allow more sensitive detection of m eans wh er ey genes with l l al e ic varia nts predisposi ng to su bstance a buse vu lnerabil ity cou ld be identified . Th is work was f accom pan ied by conti n uing work on possi ble confoundi ng eatures , including r a cia l and eth nic differences in

marker freq uencies , fou nd in association studies .

Studies of RFLP polymorphic markers at the mu opiate receptor locus fai led to reveal al lel ic association in a

. w c num ber of the same research su bjects Ho ever, the asso iation noted between su bstance a busers and " " h igher frequencies of the T a q l A a l lele of the dopamine DZ r eceptor w a s fou nd to be la r g ely d ue to u l individuals who had access to d r gs in severa d rug classes , but who preferred coca ine to O piates . PUBLI CATI O NS

' ' B F S S S B P S B i DZ R FL P a r a rs AM C utt n GR U h l . O H , m ith , ird G . e i co , uarez , g , G R Dopamin e Receptor s haplotypes and their association with substance use in black and caucasian research vol unteers . H um H ered -1 8 .

' DZ ' ‘ P AM a r a B F F S G F S U . 1 A ersico , O H , arm er , ysin R , lanagan D , h l G R Dopamine receptor gene Taq locus ' ' : o u - 4 map i ncl ud ing A 4 va ri a nt relevance for alc holism and drug abuse , D r g Alcohol Depend 3 .

- U G B K N S S . S D2 I N S 8 . hl , l um , oble E , mith ubstance a buse vulnera bil ity and receptor gen es , T

’

G P O H a r a B F Y X U G . E regor , , ang , hl R xpression and novel su bunit isoforms of g luta mate receptor genes - I u R 6 N eu r oR e ort 6 . GI uR 5 a nd G , p

P AM S S S U G . DZ c ri S I n ers ico , mith , hl R re eptor gene va ants and su bstance a buse l iability , em i nars the - Neurosciences 1 993 ; 5 z377 82 .

P AM J S SS U G rt ersico , Vanden berg h D , mith , hl R . Dopam ine transpo er gene polymorp hisms are not t - l l P 7 . associated with po ysu bs ance a buse , B io sychiatry

b S SS w J P E A P J Uhl G N . mith , N e man , vans , ickens R , Wydeven , R , ewlin D B Comor id psychopathy is not associated with i ncreased DZ dopam ine receptor T a q l A or B gene mark er frequencies in i n ca rcerated i l - s B o P l 8 . substance a buser , sycho

L A J U G SA L Da tl ossie C , Va nden berg h D , hl R , Camper . oca lization of the dopami ne transporter g en e , , on 1 3 M G - mouse ch rom osom e , am malia n enome 8 .

T er eniu U G A t . I n: J B Jomva ll r U s . L hl R ssociation strateg ies in subs ance abuse ansson , H , Ryd be g , , Vallee

B L P e 86th . , eds . roc edings of the nobel symposiu m towards a molecu lar basis of alcoh ol use and a buse ir kh a u ser rl B - S witzeri a nd : B Ve ag asel , 1 41 .

U P A . M rt : A u G s u A hl G R , ersico eeti ng Repo Collaborative pproaches i n Dr g A buse enetic , Dr g lcohol Depend - 2 .

Uh l G : DZ ! N Reply , R receptor genes the cause or consequence of su bstance a buse Trends i n eurosciences - 1 .

' r a B F DM L I B M M A KIa u n ber BA S O H a , Donovan , i ndberg , rannock T , Ricker D D , offatt C , g . ch i nd ler C , Chang

'

S K N R J U . Pr oenke ha l in : T , elson , hl G R p transg en ic mice a short prom oter confers h ig h testis expression and M l R e r od - 4 rt 8 . reduced fe ility , o p Dev

P A M W ZW B DW A N U G a rt : r . e sico , ang , lack , ndreasen C , hl R , Crowe RR Dopam ine tr nspo er gene exclusion

A J P 1 994 in . of close li n kage with sch izophrenia spectru m disorders , mer sych ; press

'

P G S M ,Y X O R e a n J P E P J L , U Gregor , aston a ng , g , Rosen DR , Tanzi R , atterson D , Haines , H orvitz H R hl G R B J G GL U R rown RH , r . enetic a nd physical mappi ng of th e S gl utamate receptor gene on h u ma n ch romosome 994 in G 1 . 21 , H u m enet ; press

d er G L a B u a M P k G B F E K J . l Elm . d nr . I n U h G R , , C , ic ens R enetic influences i n g a buse : loom , upfer D , eds 4 i P : N Y : P 1 99 n . sychopharmacology Th e fourth generation of progress . ew ork Raven ress , ; press

P AM ZW DW A U G w rt : W B N . ersico , ang , lack , ndreasen C , h l R , Cro e RR Dopa m ine transpo er gene excl usion

w r A J P 1 994 in . of close li n kag e ith schizophrenia spectru m disorde s , mer sych ; press

U . M r r hl G R olecu lar and genetic studies of the ta gets of acute d ug action , su bstrates for interi nd ivid ual I n : . M d ifferences in vulnera bi lity to su bstance a buse , and ca ndidate m echanisms for addiction olecu lar u : k M S N approaches to d r g a buse research Roc ville , D : Dept . of Health and Hu man ervices , ational I nstitute on

PUBLI CATIONS

M . c i l K . N v k f e d J . E . R . H enning , and eenan , i oti ne deli ery inetics and a buse lia bility Jou rn a l of Consulting

- 743 750 1 993 . Clinica l Psychology , ,

J H ennin field . m J . P R . . E. rf t S . S P Heishman , , nyder, and g e or ance , subjec ive and physiological effects of - 34: 1 1 - 1 8 99 . A nicotine in n on smokers Drugs a nd lcohol Dependence , , 1 3 .

O . L . e n n i W F K . S n d P . r J . H fie l J . E . P . M P A oodson , , age strom , , oland , , lade , , and g , rem ier: new nicotine n - delive r y system studies on its envi ronmental and biological impact . Jour al of Smo king Related Disorders

- 1 9 1 201 , 1 993 . SUMMARY OF WOR K

T l he prevalence of nicotine dependence , its accompanying health prob ems , a nd the difficu lty users have in t t s l T main ain i ng a bs inence com bine to make it one of the mo t prob ematic a bused drugs i n ou r society . hese stud ies were designed to provide a better u nderstanding of the pathophysiology and mechan isms of n icotine we ll ri dependence as as to cont bute to the development of putative new smoking cessation treatments .

S T everal nicotin e delivery systems have been developed to aid i n smoking cessation . wo of these , nicotine ola cril ex tr a nsd enr ra l p and n icoti ne patches , have been approved for this purpose . However , new systems na sa l are bei ng developed , incl uding a nicotine spray and a n icotine vapor inhaler. These systems a r e l r fa ctor tha notable for havi ng a more rapid de ive y rate , a t has been associated with greater potential for d . l a buse Th erefore , a though eveloped as aids for smoking cessation , such systems may be a bused i n their “ - r . T f own ig ht he first study was desig ned to exami ne the physiologi ca l , su bjective and perform ance ef ects of ri na sa l l w va ous doses of n icotine spray and vapor in haler com pared to a regu ar cigarette , h ich served as a - r . T w standa d positive control his placebo controlled , double bli nd , outpatient study il l provide needed l inform ation a bout the a buse iabil ity of these prod ucts .

A n lthoug h it is com monly thought that cotinine , the major metabol ite of nicoti ne , has i sign ificant pharm acolog ic activity -c otinine has been shown to be behaviorally active in both hu ma ns a nd other rt a nima ls . Of pa icu lar i nterest is the recent finding that cotin in e ad min istration induces su bjective changes t abstinent h uman cigaret e smoker s . The purpose of the second study is to characterize the ha nn a cod na mic t r p y effec s of cotinine in cigarette smokers unde going peri odic a bstinence , includi ng w w u ri symptoms of tobacco ithdra al , mood and su bjective state , dr g effect , per forma nce , and endoc ne

. w will effects This study , hich is in progress , provide valua ble inform ation on a neglected aspect of th e

. I n w pharm acology of tobacco dependence addition , if cotini ne is effective in suppressing withdrawa l , a ne rm w pha acologic treatment may be developed , hich could save h u ndreds of thousands of lives each year i n the U nited States . PU BLI CATIONS

a J k P e e nt E. m th e tr a tm . M E . H ennin fi eld J . I : arvi , and g . har acologica l adjuncts for of nicoti ne dependence n : i k : O O T l a d e J D . . N A M . N Y rl eans , C . a nd S , . , (eds ) icotine ddiction P r nciples and anagement ew or xford - 993 . U P . 245 261 1 niversity ress , pp ,

i fie ld B . EB . H en n n J . E . f P . B k C . w rt W Cohen , , ick o h , , un er , and g , Caf ein e a ntagonizes EEG effects of tobacco 1 - 4 P B 8. B 9 9 926 1 99 . withdrawa l . harmacology iochem istry ehavior, ,

H e in ld m K J M E . nn fi e J . E. P . I M R . M . : N . S eenan , , arvik , and g , har acology of n icoti ne dependence n il ler, .

P m o A . (ed . ) har a c l ogi ca l Therapies i n Drug and lcohol Disorders , (i n press)

E. G . e : . S M P P n fi eld J . E m ! H enni g , and i ngleton , anaging d rug dependenc sychotherapy or har acotherapy - S u 31 7 322 1 994 . C N Dr gs , ,

d . A A i fiel J . E rk ! . H enn ng , Do n i cotine replacement medications wo unique standard for n icotine ddiction - 434 436 , 1 994.

M E . H enn K in fi eld J . E P . I : M eena n , and g , . harmacologic treatment of n icoti ne add iction n iller , in i l es . Pr c N a l A M . . et (eds ) p of ddiction edici ne , (in press)

nn in l e J . S H fie d . W I J chwa ndt , and g E. ithdrawa l syndrom e: Description a nd treatm ent , nicoti n e . n : affe

a l . T r A o M P . J . E. et (eds ) he En cyclopedia of D ugs and l c hol . acm i llan u blish ing (i n press)

in L . M . H enn field . J . m S w E. P o t . I : J J et a l ch a ndt , and g har ac logica l treatment of cigaret e smoki ng n affe , . H .

E u A N Y k : M l P l . (eds .) Th e ncyclopedia of Dr gs and lcohol . ew or acm il an u b ish ing (in press)

field . J . E. L . M Henning a nd S chu h , . Tobacco dependence : Pharm aco logic determ inants . Jou rna l of Clinica l

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