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Some Aspects of Amino Acid Regulation in the Tobacco Hornworm, Manduca Sexta

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Some Aspects of Amino Acid Regulation in the Tobacco Hornworm, Manduca Sexta University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Masters Theses Graduate School 8-1981 Some Aspects of Amino Acid Regulation in the Tobacco Hornworm, Manduca sexta J. Michael Henry University of Tennessee - Knoxville Follow this and additional works at: https://trace.tennessee.edu/utk_gradthes Part of the Zoology Commons Recommended Citation Henry, J. Michael, "Some Aspects of Amino Acid Regulation in the Tobacco Hornworm, Manduca sexta. " Master's Thesis, University of Tennessee, 1981. https://trace.tennessee.edu/utk_gradthes/3008 This Thesis is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Masters Theses by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a thesis written by J. Michael Henry entitled "Some Aspects of Amino Acid Regulation in the Tobacco Hornworm, Manduca sexta." I have examined the final electronic copy of this thesis for form and content and recommend that it be accepted in partial fulfillment of the requirements for the degree of Master of Science, with a major in Animal Science. Arthur M. Jungreis, Major Professor We have read this thesis and recommend its acceptance: James M. Liles Accepted for the Council: Carolyn R. Hodges Vice Provost and Dean of the Graduate School (Original signatures are on file with official studentecor r ds.) To the Grad uate Cou ncil: I am submitting herewith a thesis written by J. Michael Henr y entitled "S ome Aspects of Amino Acid Regulation in the Tobacco Ho rn wo rm , Mand uca sexta ". I recommend that it be accepted in partial ful fi llment of the req uirements fo r the degree of Master of Science, with a major in Zoo logy. We have re ad this thesis and recommend its acceptance: Accepted fo r the Cou ncil: Vice Ch a ncellor Grad ua te St udi es and Research SOME ASPECTS OF AMINO ACID REGU LATION IN THE TOBACCO HOR NWORM, MANDUCA SEXTA A Thesis Presented fo r the Master of Science Degree The University of Tennessee, Knoxville J. Michael Hen ry August, 1981 3053788 ii ACKNO WLEDGMENTS The author is grateful for th e assistance , gu ida nce andpat ien ce of Dr. Ar thur M. Jung reis and for the mo ra l su ppor t pro vided by Jo Am He nry. also th ank Cathy W ebb for typing th is docuent m . iii ABSTRACT The nature andro les of th e hig, free amin o acid level s foun d in th e haemolymph of in se c ts is poorly un der stood. In or der to elicit aspec ts of th e reg ul ation an d me tabol ism of hae moly mph free amin o acids, we ha ve mea su re d the concen tr a tion s, hal f-l ive s, tu rno ve r ra tes and ra tes of in corpora tion in to hae moly mph pro te ins for eig,tam ino acids in th e to bacco hornwor m, Mmdu ca � immed ia tely pr ior to and th roughou t the larval-pupal tr an sformation. Throughou t th is per iod th e con centra tions of alanne, i arg in ine , glu tamate, histidine, leucne, i ly sine, proline and val ine each exh ibited stage sp ecific va riations se eming ly ind epen dent of bloo d vo lu mes an d levels of other amino acids. Hal f-lives for th ese amino acids also exh ibited repro ducible stage spec if ic var ia tions ind ic ating in de pen dent regu la tion. Half-l ives de monstra ted a wide ran ge of util iza tion (2- 170 hours) and do no t chan ge in parallel fa sh io n for th e amin o ac ids in vestigate d. Conclu sion s dr awn fro m th is stu dy are th at a) amin o acids are regula ted and b) al th oug, so me amin o acids su ch ashi stidine appear to be stored in hae molymph, mostare dy namic a lly ma in ta ine d an d mu st be ma jor de te rmin an ts of in sec t ho me o stasis. iv TABLE OF CONTE NTS SECTION PAGE I. INTROD UCTION . II. MATERIALS AND METHODS 3 Ill. RESULTS . • 12 IV. DISCUSSION 31 v. EP ILOGUE . 36 LIST OF REFERENCES 38 APPE NDIX 45 VITA ••.• 47 v LIST OF FIGURE S FIGURE PAGE I. Ty pical plot of data to the exponential decay equation (Equation 3) . 8 2. Fit of log transformed data for 1 4c-p rol in e on pink stripe + I da y to Equation 4 . 10 3. Co ncentrations of valine, alanine, leuci ne and glutamate in haemolymph of M. se xta in feeding larvae and in pharate pupae throug hout the larval -p up al transformation . 13 4. Concentrations of histidine, proli ne, lysine and argininine in haemolymph of M. se xta collected fro m feeding larvae ood in pharate pupae th "'iOiJgflout the larva l -p upal transformation . IS 5. Half-lives for proli ne and leucine in feeding fifth instar larvae and in pharate pup ae throughout the larval -p upal transformation 19 6. Half-lives for histidine in feeding fifth ins tar la rvae and pharate pupae throughout the larva l-pupal transformation . 21 7. Half-lives for valine and glutamate in feeding fifth inst ar larvae CJ'ld pharate pupae th roughout the larval-pupal transformation . 23 8. Half-lives for arg i ni ne CJ'ld lysine in feeding fifth instar larvae and pharate pupae throughout the larval -p upal transformation . 25 I. INTRO DUCTION The concentrat ions of am in o acids in tissues and haemolymph from a wide range of in sect orders exemp lified by numerous spec ies at se lected stages in deve lopment have been published (see citat ions in Flork in, 1959; Wyatt, 1961; Flork in and Jeun iaux, 1974 ). In general, these st udies have been toxonom ic rat he r than phys iolog ica l in outlook in that they descr ib e the distr ib ution of am ino acids at a single po int in time (see Sutcl if fe, 1963 ). Am ino acid metabol is m and regu lat ion in ins ects are poorly descr ibed (see however Auclair, 1959; Wh it ehead, 1969; De sa i and Kilby, 1958; Pr ice, 1961; Ch en and Bachmann- Diem, 1965 ), with few measurements hav ing been made of am in o acid fluxes throug h haemolymph or whole body pools (see Lev enbook and Dinamarca, 1966; Collett, 1976; Irv ing, Osborne and Wilson, 1979 ). The concentrat ion changes of am ino acids in haemolymph dur ing de vel o pment are record ed without references to quant ity (i.e., as the contr ibution of the blood volume to the unit bod y we ig ht ch ooged?) or met abo lic fl ux (tu rnover ). Wh ile the stat ic nat ur e of haemolymph sugar is often im pl icit in the many reports appear in g on the subject in the literature, that for free am ino acids is qu ite expl ic it . In a volume on ins ect phys iology, Patton (1963 ) writ es: ••• the va lues (of am ino acids ) for the blood of im mature stages have been se l ect ed, the reas on be in g that these va l ues are more likely to rema in constant thm those of aduIt blood •••• One char acter ist ic of am ino acidem ia in the blood of in sects is its constancy ••• free am ino acids found in the blood in the largest quant it ies are not those am ino acids essential to the nutr it ion of the in sect. From th is it can be suggested tha t the high am ino acid concentrat ion in the blood of in sects represents the storage of nitrogenous mater ia ls that can be draw n upon , accord in g to the ne eds of the tissues••• or it may ind ic ate that an exces s of am ino acids are produced from the diet and that the am ino acids are stor ed in blood unt il they can be eliminated• •• 2 This view was further supported even by Wigglesworth (1965) who, in discussing hoemolymph amino acids, included examples that demonstrate the passive nature and putative storage functions of hoemolymph: There seems to be no exact regulation of the amino acid patterns: the hemolymph of the aphid Me�uro contains the same amino acids as honey dew a in similar concentrations (Ehrhardt, 1962). If Rhodnius is fed on horse serum with added alanine, olcrune appears in the hoemolymph within a few minutes ood in the urine in half m hour, but the high level in the hoemolymph persists for weeks (Harrington, 196 1). In this study, the rates of turnover and the half-lives of eight hoemolymph amino acids were determined in haemolymph from the tobacco hornworm, Monduco sexto, immediately before and throughout the larval-pupal transformation. The questions I posed were: I. whether these amino acids ore regulated statically or dynamically (i.e., do haemolymph pools represent sinks in which levels change only with tissue needs, or ore they in constant dynamic interaction with tissues), 2.
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