Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. a eoeahat n oilpolmi eetyasta ed ob retyadesdadi unmet a is USA and the addressed urgently in be 2018). particularly to al., countries needs misappropriate et western that Ezard to in years 2016; leading recent addiction al., pain, in et Drug chronic problem (Wingo of social 2019). need treatment and dependence medical al., analgesics health the psychological et used a in and (Ruzza widely become drugs physical has liability and the opioid of potent abuse and development use most and 2008) the to the However, al., use limitation are et drug important pain. opioids treat (Robbins an severe to related human challenge is to and the severe moderate Morphine becomes in Drug and treating use 2009). problem relapse. for drug clinical al., to major of et a prompting cessation (Aguilar is eliminate, the detoxification abuse after to after relapse easy drug time of seeking not rate long for high is a craving which persist the medication, avoid to can to repeated refers relapse drug by dependence the achieved Psychological of psychological dose euphoria 1993). and the and al., the gradually dependence is et increases addiction physical and (Nestler Opioid drug repeatedly symptoms including drug withdrawal 2016). seeking mainly seeks al., compulsive et dependence dependence, Wu by Physical and 2015; characterized al., dependence. tolerance is et Zhu of which 2007a; disease, results al., brain accumulated et (Shippenberg recurrent craving and drug intense chronic and a is addiction A, Drug dynorphin BAA, Keywords: addiction. drug Introduction opioid and of brain treatment in the expression for A candidate dynorphin potential microglial a of be κ- may stimulation BBA by that dependence suggest psychological specific and and antiserum A physical signs dynorphin withdrawal morphine-induced minocycline, naloxone-induced inhibitor measured activation BAA-inhibited CA1, microglial 4 the hippocampal by and κ- staining. blocked NAc totally immunofluorescence in were double microglia acquisition and in CPP and A expression dynorphin protein induced (300 of and (30-300 expression mice BAA gene BAA the in by of stimulated and morphine injection specifically of signs, BAA subcutaneous injections 3 withdrawal Single daily acquisition. 2 morphine Multiple acquisition. attenuated 1 cellular (CPP) neuronal and results: nucleus preference and Key dose-dependently in place glia measured CA1. conditioned with were and hippocampal A level signs and dynorphin withdrawal A of NAc dynorphin staining in explored immunofluorescence and detected and Double was expression dependence biomarkers CA1. gene conditioned psychological hippocampal Prodynorphin and and mice and used. (NAc) morphine recent physical were in accumbens in signs paradigm in problem withdrawal (CPP) social inhibition Naloxone-induces preference a approach: (BAA) become place Experiment has A action. which Bulleyaconitine of disease, mechanisms investigated brain underlying recurrent study and chronic Our a is years. addiction Drug purpose: and Background Abstract 2020 1, July 3 2 1 Zhao Meng-Jing expression A dynorphin psychological via and dependence physical morphine inhibits A Bulleyaconitine hnhiJatn University Jiaotong Shanghai University Normal China East University Tong Jiao Shanghai podrcpo,ncesacmes(A) ipcma A,microglia CA1, hippocampal (NAc), attenuates accumbens BAA nucleus that receptor, illustrate opioid time, first the for results, Our implications: & Conclusions GNTI. antagonist receptor opioid 1 iY Wang Mi-Ya , 2 eMa Le , 3 hllAiAhmad Ali Khalil , 1 μ /g lottlyalvae opieCPP morphine alleviated totally also g/kg) 3 n ogxagWang Yong-xiang and , 1 μ g/kg) Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. ig hn)wt h uiyn esta 8 eemndb auatrrwt ihpromneliquid performance high with manufacturer by determined 98% than less no purity the with opioid China) of reagentsjing, treatment and the Drugs for 2.1. target methods potential and a Materials is A 2. dynorphin of dynorphin of expression abuse. expression and microglial microglial addiction that brain suggest addiction through and and dependence the A, underlying expression psychological As of mechanisms A and 2016). part the physical dynorphin al., a morphine-induced explored microglial et we of constitute (Wu involvement Finally, to laboratories the subsequent properties. in considered particularly abuse reinforcing effects, is of opioid anti-addictive further acquisition drugs BAA-induced the We CPP of with morphine effect 2005). associated reinforcing drugs, Roerig, process the and addictive test, assess CPP (Lenard other to the loss like in model weight dependence used including psychological body widely signs morphine-induced withdrawal and a inhibited induced BAA morphine-induced excretion naloxone subcutaneous attenuated fecal of whether BAA application licks, assessed psycho- which of genital and in injection physical jumps, model, morphine-induced subcutaneous mouse shakes, in a whether BAA in tested of dependence effects first physical anti-addictive We the blocks assess dependence. injection to logical aimed dependence. A we psychological bullatine 2017a). study and and al., this properties dependence et BAA In therapeutic physical potential Huang addition, have morphine 2016a; may In of BAA al., including treatment 2018). et aconitines the that (Li al., in hypothesis mice et the to and Sun lead rats A reports 2017; in These dynorphin al., tolerance et microglial antinociceptive Li morphine-induced spinal chronic 2017a; stimulating aconitines, al., through BAA, et that directly uncovered (Huang further pain al., activating studies et subsequent alleviated recent Huang and Our 2016b; lappaconitine expression al., 2020b). et and al., Li et A 2016a; Huang al., bullatine et 2020a; (Li bone al., pain of pain, et visceral neuropathic Huang and induction including 2017a; pain, hypersensitivity, diabetic without pain pain, inflammatory of antinociception pain, al., models cancer produced et rodent various China Xie bullatine (C18-diterpenoid) and in (C19-diterpenoid), 2004; in analgesic tolerance lappaconitine aconitine Nava-Ocampo, prescribed antinociceptive nonnarcotic analogs and and widely its a (Bello-Ramirez and (C20-diterpenoid) been decades is BAA four has that A it over demonstrated BAA As evidence pain aconitine, Accumulated chronic 2007). than of 2018). al., window forms various treatment et treat (Wang wider alkaloidto receptors and C19-diterpenoid opioid toxicity a lower is to bulleyanum, has binding Aconitum of of rhizomes activities the without from isolated dynorphin/ (BAA), A the Bulleyaconitine of role al., et complex Carey a 2005; suggest al., development. et reports (Beardsley These acquisition On CPP 2007). 2009). or suppressing Sullivan, self-administration by and cocaine-induced function Mysels that of 2008; reward reinstatement reported al., brain was dynorphin/ et inhibited it (Chartoff the pathway contrary, and that reward the abuse mesolimbic reported drug the also from was in release It effects dopamine rewarding 1994). repressing antagonized al., by system et monkeys (Maisonneuve rhesus reported tor in dopamine was self-administration of It and release conditions. acquisition the (CPP) certain preference under place abuse alleviates conditioned drug tion, peptides reduce 1995). opioid al., but addiction. et related effects drug the Hooke reinforcing other that 1993; of produce and al., development not et A (Takemori do the dynorphin dependence agonists, participates physical of morphine and administration of dynorphin/ system that symptoms The withdrawal anti-reward evidence 2010). growing major Bodnar, is a 1986; There also al., et is (Pfeiffer pathway Bruchas functions 2007b; receptor al., neuroendocrine et and Shippenberg emotions dynorphin/ 2004; ature, the al., different et and (Yuferov with 2010), system subtypes al., nervous central et opioid the three in to distributed binds widely also A dynorphin to system, especially nervous central affinities, the in distributed Widely κ- podrcpo gnssehleoylzcn n 5,8 teutdcciebhvoa sensitiza- behavioral cocaine attenuated U50,488 and ethylketocyclazocine agonists receptor opioid κ- podrcpo ciaini ri.Orrsls o h rttm,ucvrta A inhibits BAA that uncover time, first the for results, Our brain. in activation receptor opioid κ- podrcpos(alnadLsi,18;Shazr 2009). Schwarzer, 1986; Leslie, and (Fallon receptors opioid ulycntn BA a ucae rmZln i-hraetcl(Nan- Bio-Pharmaceutical Zelang from purchased was (BAA) A Bulleyaconitine . κ- κ- podrcpo ytmpasa motn oei anaagsa temper- pain/analgesia, in role important an plays system receptor opioid podrcpo naoit o-N n rdnbokdstress-induced blocked arodyn and nor-BNI antagonists receptor opioid κ- podrcposi ait frdn oeso anhypersensitivity pain of models rodent of variety a in receptors opioid 2 κ- podrcpo ytmi h rgabuse drug the in system receptor opioid κ- podrcposare receptors Opioid κ- podrecep- opioid κ- κ- Opioid opioid Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. h CDcmr n nlzduigteEhVso T80 h odtoigsoewsepesdb the by by recorded expressed was was score chamber conditioning each The in for spent 8.0. compartments XT time three 2020). 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(Marszalek-Grabska striped 40 were with The baseline white cm) x decorated exclude and was to black 40 25 2019). chambers adjusted with x x was main al., one 5 the (25 et other of x chambers the Shi one 25 equal-sized other, 2008; compartment, each Two (null al., distinguish chamber et compartments. protruded (Bahi three white rewarding of drug minutes consisted of 30 test for effects paradigm. recorded the and and explore observed apparatus were an CPP which by seventh weight, jumps, followed 2.4. the shakes, by body included On a.m. of mg/kg) signs 10:00 loss withdrawal 100 days. injection. and at The naloxone and consecutive excretion mg/kg) later. after 80 6 fecal (100 hours and for 4 40, morphine licks mg/kg) of 20, (5 genital p.m. injection naloxone 10, 4:00 of subcutaneous (5, and injection single 2019). intraperitoneal doses a al., a.m. received escalating et 10:00 mice Bobzean with at day, 2011; mice injections al., in et subcutaneous administered (Goeldner twice-daily established was previously morphine as Briefly, University performed dependence Tong was physical Jiao dependence morphine Shanghai physical of of Induction randomly experimental Committee and were Welfare 2.3. conditions group) and housing per Care All (n=10-12 Animal manner. Mice blind China). the a (Shanghai, by experiments. in approved the performed were were before room tests procedures days standard behavior at 3-5 the water to and and assigned acclimatized food to were access mice a free in All with maintained p.m.) were a.m.-7:00 animals (22 7:00 The temperature period China). (light (Shanghai, cycle light/dark Sciences 12-hr Biological All for Institute saline. 2013). Animal normal al., validatedExperimental 0.9% et animals also in Yamada Experimental was diluted 2010; or al., specificity 2.2. dissolved et Its (Wakabayashi were laboratories reagents sheet. other and data from drugs manufacturer’s test the the absorption (0%), to antigen B antiserum the according dynorphin anti- to by The polyclonal (0%) not USA). leu-enkephalin rabbit but CA, (100%), or The (Burlingame, A (0%) USA). dynorphin Pharmaceuticals to Phoenix Mo, specific from was Louis, purchased serum naloxone was (St. and A Sigma-Aldrich (GNTI) dynorphin 5’-guanidinonaltrindole Sinopharm neutralizing from Both and obtained respectively. China), were China), (Shanghai, hydrochloride (Shanghai, Biotech the Yuanye Co. from Reagent China), purchased Chemical (Shenyang, were Group sodium pentobarbital Pharmaceuticals and Northeast minocycline, hydrochloride, Morphine chromatography. ± )i h hnhiJa ogUiest xeietlAia etr(hnhi China). (Shanghai, Center Animal Experimental University Tong Jiao Shanghai the in 2) aeautSismc 89wes 02 )wr ucaefo h Shanghai the from purchase were g) 20-25 weeks, (8-9 mice Swiss adult Male . P a oiinpeeec o ieetcabr n sue to used is and chambers different for preference position a has CPP h opieamnsrto aaimfridcinof induction for paradigm administration morphine The . 3 β- nopi (0%), endorphin α- neo-endorphin Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. b-- FP rNu-muooiieclsi AS n ipcma A,teiae eeacquired were images in the A CA1, in dynorphin hippocampal brain of and the intensity brain NAcSh relative to the in the according to quantify cells mm, according To NeuN-immunopositive mm, 0.98 -1.94 or microscope. to visualized to GFAP- confocal mm was mm Iba-1-, a NeuN 1.70 -1.46 and under bregma bregma GFAP, (from coordinates) (from Iba-1, microglia, CA1 stereotaxic A, (NAcSh) for hippocampal dynorphin accumbens and antibody of an- nucleus coordinates) secondary Expression secondary stereotaxic of anti-rabbit anti-mouse 2018). shell goat goat al., the Alexa-555-conjugated et Alexa-488-conjugated PBS, in the (Qi the with with neurons washing and 37 or After A at astrocytes neurons. hour dynorphin for 1 antibodies for mon- Millipore) for tem- primary mouse tibody polyclonal; incubated room (1:100; applied mouse were markers: GFAP (1:60; at The sections microglia, cellular NeuN hour the for and and antibodies. Germany) 1 Pharmaceuticals) astrocytes Darmstadt, primary for for Phoenix Millipore, with (v/v) oclonal) polyclonal; monoclonal; hours X-100 rabbit mouse 24 Trixon (1:100; (1:100; for 0.5% Iba-1 antibody 4 and A di- at (v/v) dynorphin embedding 30% incubated serum included temperature and then cutting goat optimal 20% and 10% then the (10%, in perature was in Paraformaldehyde solution incubated embedded 30- were sucrose was The into 4. brain gradient sections cut at paraformaldehyde. dehydrated and with hours The 4% Microsystems) dehydrated (Lecia 12 of 4. was agent for at mL brain paraformaldehyde PBS) 20 the with 4% by luted and followed the PBS saline, in mL/kg), with 0.9% fixed (5 removed of and sodium dissected mL pentobarbital was 20 1.5% brain with intraperitoneal confocal mi- SP8 by perfusion with TCS anesthetized method intracardially a deeply published using and were previously out Mice the carried to was modifications. according hippocampus nor Germany) and Wetzlar, NAc Microsystems, in (Leica neurons microscope and astrocytes, et microglia, of (Leitermann ers manual operation the staining and 2.8.Immunofluorescence 2012). to al., China) according (ELISA) et assay USA) Jiangsu, determined Nocjar were immunosorbent Biotechnology, CA, 2004; enzyme-linked hippocampus Burlingame, al., of fluorescence and Pharmaceuticals, Institute NAc commercialized (Phoenix (Beyotime in using kit concentrations assayed assay protein at was protein centrifuged total A and acid The dynorphin 7.4) seconds bicinchoninic (pH minutes. 15 Tris-HCl 15 standard for mM for rpm 10 by 4000 in 4 was at China) at change homogenized Shanghai, rpm were Co, fold 1500 Tissues Equipment The (Fluko measurement. homogenizer further 2017). a until with al., -80 an et stored (Reiss and A nitrogen gapdh dynorphin for was of ATG Measurement which GC TCC TCA GTG 2.7. prodynorphin, AGG TGA for CCA TTC AC 2 and the GTC GGA USA); Francisco, using ACA GTG San calculated TCC GCT ATG Biosoft, were and AGG Premier sequences ATG AC 6.0, primer TTA (version ACG using reverse and 6 Real-time Germany) and Premier TC forward Hamburg, by manufacturers. CCT The designed (Eppendorf, CAT the Japan). realplex Toyobo, CGC by I, ep AGA provided Green ATG Mastercycler RT-qPCR (SYBR instructions a ReverTraAce Mix the with Realmaster the to the performed using according was cDNA reagent Japan) PCR into TRIzol the Osaka, quantitative transcribed using Co., reversely mice (Toyobo of were hippocampus kit and and USA) (NAc) accumbens Carlsbad, nucleus (PCR) (Invitrogen, reaction from 2005). chain isolated Animals polymerase Roerig, were over quantitative and RNAs andreal-time slowly covered. total transcription, Lenard to administrated reverse was 1980; extraction, was applied cannula Wilcox, RNA drug was and of 2.6. The cement (Hylden cap recovery. 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No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. yitaeioelijcino aooe( gk, 0mntsatrsbuaeu neto.Subcutaneous injection. subcutaneous after minutes precipitated 40 were mg/kg,) signs (5 Withdrawal naloxone injection. of intracerebroventricular injection first intraperitoneal the by post intrac- minutes (6 30 first saline was the injection 1) 6 received of dilution, group) (1:30 injection antiserum per μ subcutaneous A (n=10 second dynorphin mice 2) the dependence mL/kg), by physical followed morphine injection of erebroventricular groups four Additional BAA- systemic test). restored (P weight completely signs dependence. nearly withdrawal body physical suppressed minocycline and in morphine-induced intracerebroventricular excretions shown influence attenuated with fecal to significantly pretreatment As licks, However, failed mice genital minocycline injection. dependence intracerebroventricular jumps, physical subcutaneous precipitated whereas shakes, morphine loss; after were including in minutes signs BAA signs 40 Withdrawal withdrawal of naloxone-induced mg/kg) injection injection. subcutaneous (5 intracerebroventricular 6A-6E, naloxone first Fig. of the injection post intraperitoneal hours by subcutaneous 4 second was the injection by followed (6 injection intracerebroventricular saline (6 first 1) mice dependence the of physical received injection morphine group) of groups per Four (n=10 separately. 2013), injected al., intracerebroventricularly morphine and et were 2016) Kobayashi BAA-inhibited al., 2002; 2013) and et al., brain (Li et (Wu antiserum in minocycline A A inhibitor dynorphin dynorphin activation microglial of the expression dependence, dependence microglial physical morphine the BBA-inhibited between mediated (Fig. A relationship A/NeuN dynorphin causal dynorphin of expression or microglial 5N) (Fig. Brain (Fig. 3.4. above A/GFAP as dynorphin mice not on but same BAA 5M), (P the of Fig. 5O). 1.9-fold from effects by t-test; CA1 A/Iba-1 stimulatory Student dynorphin two-tailed specific hippocampal of and intensity in same immunofluorescence observed the increased Furthermore, were with 5A-5L), expression 4O). 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All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. yial hksadbd egtls ihED with to loss 30 weight from ranging body responses allevi- doses and BAA jumping In shakes of induce typically bi-daily mice. injection not dependence subcutaneous contrast, physical did single morphine In addition, BAA 300 in In signs subcutaneous withdrawal 1986). loss. daily naloxone-induced al., weight which ated et (Tang in body challenge finding and nalorphine previous following 300 excretion the withdrawal to fecal induced with up naloxone licks, consistent BAA of application of genital and injections and subcuta- jumps, days, subcutaneous physical bi-daily consecutive shakes, 7 following composing for developed including addiction mice dependence signs in induces physical morphine study, narcotics of present injections opioid the neous other In and dependence. morphine psychological of application Long-term test). 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All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. eto H ta. 02 a ta. 05 ue l,21;W ta. 08.Orpeetsuyprovides study present Our 2018). dependence al., drug et attenuate Wu to 2017; A al., dynorphin et state express Wu activation to 2015; alternative microglia stimulates al., an antinoci- have BAA et and Fan neuroprotection that microglia exhibits 2012; evidence that and al., additional shown cascades have et anti-inflammatory (Hu studies reward activates Verkhratsky,ception recent which dopamine and neural hand, Garaschuk state, the 2012; in protective other regulate (Kovacs, or changes the addiction ultimately chemical and On and dependence produces reconstruction, neurons, drug 2019). and synapse enhance apoptotic microglia affect and activates al., of pathway which et abuse phagocytosis signaling (Miguel-Hidalgo Drug factors, and alcohol inflammatory 2012). transduction and Hutchinson, of opiates signal and number as Coller such influ- large 2009; drugs are a abuse al., hippocampus of to and et VTA models exposure Hutchinson NAc, rodent chronic 2002; in the by particularly in activated brain 2018) and in al., pain. enced microglia et visceral that (Sun and indicates lappaconitine evidence pain Cumulative and inflammatory 2016) pain, al., cancer et bone (Huang pain, A neuropathic bullatine specific 2020b), the other al., of and et addition GNTI rats agreement, in that self-administration In and fentanyl the of 2008). 2014), decrease al., al., agreement proportion-dependent et et a in (Negus (Freeman produced are fentanyl monkeys to results in U69,593 agonist remifentanil The and the cocaine dependence. that of psychological publications previous and the physical with the morphine addition, In BBA-inhibited acquisition. systemic CPP antiserum and A 1993). signs specific dynorphin withdrawal al., the highly morphine et of BBA-inhibited injection (Takemori injection intravenous systemic intracerebroventricular dependence eliminated single that physical totally that demonstrated morphine further reported of study previously symptoms present was withdrawal The route It attenuated cerebroventricular A the hippocampus. through and dynorphin injections NAc of intervention around following located the is by the which supported lappaconitine to further and parallel is notion A are The bullatine results BAA, The not injection, of but after effects. injection dynorphin hour anti-addictive subcutaneous 2016b). of 1 its and expression at of intrathecal the hippocampus time-course which stimulated and in the NAc the in findings with whether A morphine- agreement previous dynorphin explored in in BAA we of was of study, expression which injection this the Subcutaneous In stimulated mice dependence. 2016). psychological treated al., and dependence et physical (Assar model morphine scores CPP conditioning the antagonists morphine A/ receptor in dynorphin in D1/D2 especially of decrease the administration a memory of intra-CA1 in microinjection The and and 2013). learning sulpiride, Hippocampal al., lacking and of 2007b). et SCH23390 mice al., Riahi reward-association et 2010; in in Shippenberg Dong, 2005; increased and demonstrated al., (Fanselow are et been (Chefer NAc abuse has of in CA1 drugs McGinty, opioid levels of and dopamine reward (Meshul the dynorphin/ liability Extracellular counteracts the abuse on drug that NAc 2009). in acting revealing implicated driving al., receptors, by been for et have and neurons Volkow NAc that important dopamine VTA that 2000; and is seen addiction. of cortex be produce (NAcSh) activity prefrontal can to , it drugs NAc the Thus, NAcSh for of sites regulates 2018). important in shell al., A are et 1996; release area, the Dynorphin cerebroventricular (LeGates al., the dopamine behaviors to around reward et stimulate in input located (Murphy ventral role effectively hippocampus, VTA Hippocampal a from into drugs play may projecting locally Opiates which 2016). pathway or activity intracerebroventricularly al., dopamine 2012). administering et mesolimbic after al., Sebastian the hour et at 1 (Patyal within act NAc NAc heroin, to and (VTA) alleviates morphine markedly tegmental it as moreover, such acquisition, Opiates CPP dependence. and psychological subcuta- signs and Single withdrawal physical induce both scores. not morphine-induced conditioning does high BAA with that (300 acquisition indicate BAA CPP of remarkable injection induced neous days consecutive 5 for μ- or δ- podrcpo naoit lce h nioietv ffcso A L ta. 06;Huang 2016b; al., et (Li BAA of effects antinociceptive the blocked antagonist) receptor opioid κ- κ- podrcpo ytmi A n ipcmu a lsl soitdwt BAA-attenuated with associated closely was hippocampus and NAc in system receptor opioid podrcpo naoitGT,gvnitaeervnrclry loetrl eliminated entirely also intracerebroventricularly, given GNTI, antagonist receptor opioid μ /g nieyaoihdmrhn-nue P custo.Alteeresults these All acquisition. CPP morphine-induced abolished entirely g/kg) κ- κ- podrcpo gns avnrnApnse self-administration punished A salvinorin agonist receptor opioid podrcpo ytmihbt oaienuornmsinand neurotransmission dopamine inhibits system receptor opioid β- κ- nopi ntesia od(ie l,21a ie al., et Li 2016a; al., et (Li cord spinal the in endorphin podrcpo gns 5,8Hit AS,resulted NAcSh, into U50,488H agonist receptor opioid 8 κ- podrcpo naoitnrBI(u not (but nor-BNI antagonist receptor opioid κ- podrcposi mesolimbic, in receptors opioid κ- podreceptor opioid κ- opioid Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. ramn o loo n rgadcin vlainadpormpann 31:113-123. planning program and receiving women Evaluation in addiction. responses of coping drug and reinstatement and coherence, of alcohol the sense Spirituality, for of (2008) treatment H mechanisms Amaro G, Prado Neurobiological 59:253-277. S, Arevalo reviews (2009) research J Brain Minarro preference. place M, drug-conditioned Rodriguez-Arias MA, approved and Aguilar read authors All paper. experiments; the the wrote References MJZ performed and KAA YXW and and manuscript. LM data; final MYW, the the analyzed experiments; YXW the and designed LM and MJZ, conceived MJZ and YXW work. this in contributions interests Author financial competing no declare authors (#81673403) The China of Foundation (16-K01). interest. Science Disorders of Natural Psychotic National Conflict of the Laboratory from Key grant Shanghai a a the by and a is supported is BAA was A study that dynorphin This indicate of results secretion these and treatment. expression all addiction psychological Acknowledgements microglial drug together, for targeting opioid Taken especially the that addiction, addiction for suggest 1994). opioid approach opium also treat potential Qu, or data to and heroin The candidate BAA of (Qu development dependence. duration the clinical years whose promising of 4 and patients administration to unique addict morphine oral tears, 1 drug shedding both addition, yawing, from in alleviate as In such ranged restlessness to symptoms withdrawal and BAA dependence level. eliminated mydriasis for psychological or animal chilliness, reduced base especially days the et pharmacological 6 addiction over at Arevalo solid opioid lappaconitine dependence 1983; a analog solve provides psychological al., to study and et present current physical (Woody few at Our substances relapse approaches a addictive euphoria, On limited. to for the only produce are carving leads Thus, However, 1993). still eventually not are DiGeorge, 2008). which conceptually. patients does many al., and symptoms, emphasized and and withdrawal (Wilson be it, addictive improving use undergo should not after to clinical addiction adrenergic willing is its unfortunately for of are it limits psychotherapy patients activation hypotension since respect, via postural treatments Clonidine other symptoms management of other 2017). the withdrawal well-equipped side-effect al., to et diminishes the in superior Tran although and inpatients 2015; is Jarvis, agent to and and detoxifying provided (Kampman receptors, non-opioid just dependence most physical a is is own substitution approachis however, its treatment methadone to treatment, main and The due symptoms, This institutions 2007b). withdrawal al., relieves drugs et used. and taking (Shippenberg detoxification commonly and cause is seeking be addiction compulsive could drug by it characterized for effects highlight disorder adverse relapsing the results hypersensitivity. and of pain These cytokines chronically regardless and proinflammatory a 2020b). dependence to is psychological al., contrast addiction and et in Drug physical of A, Huang morphine models dynorphin 2018a; inhibits rat releases analogs and al., the and neurotrophins, et its in expresses and Sun minocycline and microglia of 2016; of BAA injection stimulation al., of intrathecal that et by effects (Huang blocked antinociceptive also hypersensitivity were the pain morphine-induced lappaconitine Consistently, A of and BAA-inhibited injection bullatine A systemic intracerebroventricular dependence. bullatine analog blocked that psychological BAA entirely demonstrated cords and the further spinal minocycline We the physical of inhibitor in 2016). activation injection A al., dynorphin which microglial et of the in expression (Huang NeuN. finding neural) rats marker or previous neuropathic neuronal astrocytic of the not and staining GFAP (but and with immunofluorescence marker microglial by microglia agreement stimulated astrocytic identified in in specifically Iba-1, CA1, are only marker hippocampal cellular results expression and microglial The A with NAcSh dynorphin A in stimulated dynorphin neurons of BAA and of astrocytes in injection not Subcutaneous addiction. and 9 α 2 - Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. aslwM,Dn W(00 r h osladvnrlhpoapsfntoal itntstructures? distinct functionally hippocampus ventral and dorsal microglial the spinal Are stimulating 65:7-19. (2010) glycoside Neuron by HW iridoid pain Dong effective MS, neuropathic principle Fanselow the reduces methylester, rotata, Shanzhiside Lamiophlomis (2015) herb YX β- Wang analgesic N, the Gong Journal from TF, The Li brain. rat H, the Fan managed in peptides A enkephalin 249:293-336. (2018) and neurology dynorphin K comparative of and Dolan Distribution of Drug (1986) M, FM use. Shanahan Leslie JH, alcohol CA, Fallon non-beverage Day 1:S184-s194. and L, Suppl cost-savings Burns 37 Acceptability, E, review Australia: alcohol Baldry Sydney, B, in Clifford program Phar- alcohol ME, Cecilio dependence. drug N, of Mech- Ezard treatment abuse: and of 134:219-245. prediction drugs for therapeutics by approaches caused & signaling therapeutic macology immune new central kappa-opioid and of 25:5029-5037. of mediators Endogenous Implications Neuroscience Journal anisms, (2012) (2005) for The MR Society TS Hutchinson the cocaine. JK, Shippenberg Coller of to JE, journal vulnerability official Pintar and the J, dynamics : dopamine Moron neuroscience mesoaccumbal EA, regulate in Bolan systems T, cocaine receptor Czyzyk of selective VI, effects Neuropsychopharmacology the Chefer of molecular to College and Exposure American the (2008) behavioral of the publication Jr. 33:2676-2687. official WA, modulates : Carlezon A Neuropsychopharmacology BM, salvinorin rats. 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(2019) impairment Psychoneuroendocrinology D2-like reinstate- S stress-induced Chauvin period. and and for M, preovulatory accounting acquisition Lombes D1- during expression S, the Mhaouty-Kodja (2016) dynorphin V, in hypothalamic Grange-Messent A involved 312:394-404. stimulate F, research are Haghparast Billan brain Le hippocampus Z, Behavioural M, Ayrout the Fatahi preference. of place MH, conditioned region Farhadi morphine-induced CA1 of A, the ment Farhoudian in receptors D, dopamine Mahmoudi N, Assar nopi xrsin Neuropharmacology:S0028390815301027. expression. endorphin 10 Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. oaah ,IaaaS hooiT iaoK ciuaK aaooK iaaaA aeciH, Takeuchi A, Hirakawa microglia. of K, polarization Sakamoto M1 K, inhibits selectively Uchimura Minocycline K, (2013) K Hirano Kadomatsu T, N, Ishiguro Ohgomori A, S, Suzumura Different 42:3321-3330. Imagama for research Evidence K, Neurochemical Preference; Kobayashi Reinstatement. Place and Conditioned Period Morphine-Induced Extinction of of Period Characteristics Extinction Reinstate- Attenuated not Agonist but Cannabinoid Intra-accumbal ment (2017) A 149:212-219. Haghparast extinction H, behavior reduced Khaleghzadeh-Ahangar & blockade Physiology receptor morphine. CB1 of Intra-accumbal reinstatement (2015) and A medicine Haghparast addiction of H, Journal Khaleghzadeh-Ahangar Use. Opioid Involving Guideline Addiction Practice of National Treatment (ASAM) the in 9:358-367. Medicine Medications Addiction of of pharma- Use Society the of American for journal (2015) European M Jarvis technique. 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Dis Death Cell 12 Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. agX,LuX,WiHaL,Wn D iLn I(96 TDE NTEAAGSCACTION ANALGESIC THE Sinica. ON Pharmaceutica STUDIES Acta (1986) A. LI BULLEYACONITINE Ai-Ling OF MD, DEPENDENCE Wang PHYSICAL LU, Wei-Hua AND XJ, Liu of XC, pharmacology Tang of peptides Journal A The [des-Tyr1]dynorphin 266:121-124. mice. and therapeutics morphine-dependent A in experimental dynorphin tolerance and by and withdrawal Suppression opiate (1993) of expression NM the Lee HH, expression. Loh A AE, Takemori dynorphin C18-diterpenoid spinal a of Lappaconitine, stimulation (2018a) through Y-X Wang 235:2559-2571. pain X-Y, Psychopharmacology chronic Li T-F, in Li antihypersensitivity Q, place exhibits Huang alkaloid, conditioned Y-R, Wang morphine-induced J-P, the Ao of 379:112349. M-L, in expression Sun research receptors and brain orexinergic acquisition Behavioural of the Role rats. in (2020) in A hippocampus preference Haghparast the F, of Motamedi gyrus S, Phar- dentate Jamali K, addiction. RayatSanati drug M, of Shirazy pathophysiology the and 116:306-321. Dynorphin therapeutics (2007b) & VI Phar- macology Chefer A, addiction. Zapata drug TS, of Shippenberg pathophysiology the and 116:306-321. Dynorphin Therapeutics (2007a) & VI Chefer macology in A, Zapata GSK3beta 371:339-347. TS, Accumbal therapeutics Shippenberg of experimental Memory. Roles and Object Differential pharmacology and (2019) Aversion, of EM Place Journal Unterwald U50,488H-Induced The C, Preference, con- Wolsh Place E, ethanol Morphine-Induced Biology. Weltin versus inhibits Addiction von Cocaine Oxytocin JL, accumbens. Barr (2016) nucleus X, S. the Shi Iain, in diseases. release Bohrer, neuropsychiatric dopamine Kathrin, ethanol-induced in Bowen, and T., functions sumption Michael, their Peters, on T., insights Sebastian, dynorphins–new 123:353-370. of therapeutics years counter- & cebranopadol 30 Pharmacology of 112:101-105. (2009) action Peptides agonist C dependence. NOP Schwarzer physical (2019) Annals G promote Calo to brain. C, liability the Trapella its of EC, acts Gavioli systems VA, memory Holanda 1141:1-21. the C, Sciences and Ruzza of addiction Academy Drug York associating (2008) New in BJ the receptors Everitt of orexin-1 KD, Ersche hippocampal 24:237-248. TW, dorsal pharmacology Robbins of Behavioural Role England) stimuli. (2013) contextual (London, A with pain reward Haghparast of morphine F, journal NO: Khodagholi opioid Patent E, European delta Riahi US and sensitivity. Mu (2017) In: colonic C Gaveriaux-Ruff increased G, addiction. Dietrich show 21:623-634. F, treat mice Barreau J, to knockout Boue medication T, receptor Secher Chronic a RA, (2018) Ceredig as S D, Wu used Reiss accumbens. W, derivatives Wang nucleus Aconitane C, the (1994) Luo R, of P 5,290,784. Li terminals Qu H, Y, presynaptic Liu G, Qu in Cai vesicles T, Sun receptors. glutamate 14:1744806918781259. M, opiate pain the Wang Molecular kappa H, decreases Yao by K, pain mediated Ren inflammatory Psychotomimesis B, Guo (1986) C, HM Qi Emrich 233:774-776. A, NY) York, Herz (New in V, Science concentration Brantl 6:82. dopamine neuroscience the A, terminal behavioral in modulates Pfeiffer in orexin hypocretin-1 Frontiers and Local shell. dynorphin (2012) accumbens dimin- SL of nucleus shows Borgland the and depression EY, system, of Woo dopamine model R, the Patyal animal of defeat regions social 218:138-153. mesocortical The Neuroscience in (2012) hypothalamus. J orexin Panksepp of P, levels Feng ished J, Zhang C, Nocjar 13 Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. hn ,SiYG od ,Wto ,K - 20)Cnrlkpaoii receptor-mediated kappa-opioid Central (2007) M-C Eu- Ko studies. S, expression mRNA Watson BDNF 570:89-96. J, and pharmacology Re- of behavioral (2004) Woods journal ropean MJ nor-Binaltorphimine: Y-G, Kreek of with J, effects haplotypes Shi antidepressant-like Ott of association SM, H, and Leal structure Zhang A, (OPRK1) Ho gene 14:793-804. D, receptor Pharmacogenetics Gordon opioid addiction. kappa DA, opiate human Nielsen the KS, of LaForge definition journal D, Tohoku Fussell The Impairs V, Amygdala. Transiently the Yuferov Disharmony in Levels Occlusal A 230:49-57. (2013) Dynorphin medicine Increasing M by experimental Onozuka Mouse bulleyaconitine of T, the of Yamamoto in effect Memory KY, and therapeutic Kubo Learning for Y, Mechanisms Ono (2018) 14:1744806918797243. K, Yamada XG pain Liu Molecular Morphine- BT, pain. (2016) Wen chronic J RP, Wang on Pang Y, A HQ, Ma extinction. Zhu AE, and Ryabinin insula MX, L, of Xie Yang 131:192-200. inactivation W, Memory to Zong interleukin-10 Resistance and J, Learning monkeys: Spinal Wei of rhesus C, (2018) in Neurobiology Liu YX preference C, place Wang Li conditioned F, XY, induced Bai Li N, Zhao L, X, Wu Hao Immunity:S0889159118302319-. A, & Evhy Behavior A, Brain microglial through Usman roinflammation. neuropathy XQ, in Tang antinociception XF, produces Mao Peptide- HY, 37:11701-11714. Glucagon-Like Wu Neuroscience Mediates for Interleukin-10 Society Autocrine the (2017) of Jacksonlewis YX Microglial journal Wang 1-methyl-4-phenyl-1,2,3,6- C, Spinal XF, the Receptor-Induced Vadseth Mao in 1 K, XQ, neuroprotective Tang Tieu is HY, 22:1763. Psy- activation Wu S, disease. (1983) microglial Parkinson Przedborski of A of M, model Vilaperell´o Blockade Hole mouse (2002) I, H, tetrahydropyridine DH Herman 40:639-645. Ischiropoulos psychiatry Choi J, P, general V, Blaine of Teismann AT, Archives D, Beck help? CP, Wu it Does O’Brien Animals: AT, on addicts. and Society opiate McLellan Humans the for L, chotherapy of in Luborsky journal Addiction official GE, Drug the Woody and : Seeking pharmacology Novelty neuroimmune (2016) 11:456-470. of Pharmacology MD Journal NeuroImmune Li Molecules. JS, to Choi Behavior From T, detox- Nesil opiate T, in 10:529-535. alone Wingo treatment clonidine abuse versus substance clonidine displays of with combined plant 107:82-90. Journal Methadone Anesthesiology aconitum ification. (1993) from vivo. WS isolated in DiGeorge RS, and A Wilson vitro Bulleyaconitine for in (2007) Society properties GK the Wang anesthetic of SY, local journal Wang long-acting official gonadotropin- P, the Gerner pulsatile : CF, driving neuroscience Wang activity of Journal neural The of arcuate goat. 30:3124-3132. the oscillation the of Neuroscience neurons in H, periodic Tsukamura kisspeptin secretion of in Y, hormone A Mori generation releasing dynorphin DK, and in Clifton B VM, Neurokinin participate Navarro (2010) nucleus K, H secrete Okamura Mogi RA, and S, Steiner Ohkura produce K, K, Maeda astrocytes Murata and Spinal T, Nakada (2013) abuse Y, Wakabayashi V drug Hook in 47:109-115. T, role Neuropeptides Yaksh dopamine’s B, neuropeptides. Fitzsimmons dynorphin Imaging L, (2009) Funkelstein F A, Telang Wahlert 1:3-8. R, Suppl Baler 56 GJ, Neuropharmacology for Wang 37:824-839. Naltrexone addiction. Pharmacotherapy JS, and Women. Fowler Buprenorphine, Pregnant Methadone, ND, in Volkow (2017) Disorder TJ Use Todd Opioid KM, Vest of Treatment RH, Stone the BL, Griffin TH, Tran β- nopi xrsin h ora fnuocec h official the : neuroscience of Journal The Expression. Endorphin 14 β- nopi xrsin eaae rmantineu- from separated expression, endorphin Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. μ multiple 250 in (300 CA1 bar: hour BAA 1 ( hippocampal or Iba-1 obtained in mL/kg) marker were ( neurons A/microglial CA1 (10 dynorphin or hippocampal saline with of of astrocytes stained sections injection in Frozen subcutaneous not mice. after but mg/kg)-treated (10 microglia, morphine in twice-daily expression A dynorphin 5. Fig. t-test. 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ImageJ the using magnification M # P ,dnrhnAGA ( A/GFAP dynorphin ), < .5 oprdt h aiecnrlgopadBAgroup, BAA and group control saline the to compared 0.05, B n selective and ) F-J 16 ± n selective and ) ...(=0prgop.*, group). per (n=10 S.E.M. κ- N podrcpo naoitGT ( GNTI antagonist receptor opioid n yopi /eN( A/NeuN dynorphin and ) κ- podrcpo naoitGNTI antagonist receptor opioid # ± P < ...(= per (n=6 S.E.M. .5 compared 0.05, O were ) C ) Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. Data may be preliminary. i.4 Fig. 3 Fig. 17 Posted on Authorea 1 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159363388.81912664 — This a preprint and has not been peer reviewed. 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