ON TESTIS AND EPlDlDYMlS OF RATS - Pondicherry University ...

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0.1 00 01 " 0.8 I . .lo Dl* rcnn IW *c wl ~*rbtl I:#* ? lin'iw or rrDo on I& nnimty d mlah in ~hc ~llll~el~nJri;~l and micmaml fmh nf nt lalb. Thc ;rlirilh.i .In cxpn%ud tn pmllmin pr mp pmKin 11 35 .C. I Ihr V~IIIM IR. PIF%~(R( ns mn f SD Inr six mntd anl is cr~riltrnl~~l iu~ltn~ill. rn #mu*. 'Pz0.05 hmnridcltd lo h .~pmik.!nl npilm.il mnlml numbcr or Lcydip dl. germ cell and steroiilogcne enzyme activity. as well na impaired Lcydip ell LH mponsivencs (Mably el al., 1992: Joli~tw~n ct 211.. 1992). fhc weight or the epi. didynlo und u~crssory sex orgsns rquirc r continuous rlndrogcnic stimulation Tor Ihcir normal gro~vll~ and l'u~~u~io~~s (Kli~ulcltcr und Ha. I'FlUt. 7'hc wcigllts or u-ry sex organs have hrn uwd its b~wswy or andrugens (Milrkur and I lc I I IILY~ or 1'c'1)1) Im ~ kwh d hydm(m Fox& .... f,.,., ,,, IIIC IIIII~I.~IIY~~~~:#~ awl oranwwlu!l rt.aliutr ,,,,,.,,, IIK ,.l~lr. rtr srl~r\w+~l ~knwl lwn I-r nbp tn8*r?8l .,I I* '1 .Is I ~ !\I) K ~ rlv $1, ~ conlrd ~ ~ and us crpmlrltlsl ,1,1. ~r Fvq,p .I'rllll5 n ~~,n~nlmrl In k stfnikionl ,1. l,,,.l 1111111111 4.4. Elrm dTCDDon lk*wkdUpidpraiiblbnin milochondri~l and miaolavl tnnia d N W. The nlua mn apnrd in Whh w mt pmccin nl I5 T as rmn f SO Icr ua mnlml and Sia riprirrnUl anirmls pl mp. 'P < 0.05 nl ranidnal lo be sbiiknt .pin. mnrml. Chattopadhyay. 1982). TCDD has ken reported to reduce plasma tcatwlerom. Sadihydrotcatosterone (DHT) and LH (Mably el al.. 1992). The observed reduction in the weights of ~k amnory su orpm rdkts reduad bioavailability and/or production of andmgm in the TCDD-lrci~tcd MIS. TCDD has bccn shown to ad through Ah mptor. a lipnd aninled nuckar lrnnrriplion raclor (Safe. 1990). The nuclear Ah wptor conrplea is a heterodimcr conlnining the Ah receptor und Ah naptor nuclear lrnnsloetltor prolcinr and the molcculur mechanism or Ah +or sction has bem shown to be assodated with bindinp or \he Merodimcr lo dioxin mponsin ekrmts in rcgulr~ry regionn of Ah-mponsive gene (Sure. H)OI). It hu bacn repod that TCDD and its mngnm bind to the Ah mqlor, which mulls in the induction of m m I enzyme rysremr that may be mpomiMe Tor the production or rcaftive intmndnta such 1s Tree rndbh IKimbmu~h. 1974: Neben a 01. 1981: Slohr. I WO: Slohs el ul.. 1991). The mechanism of TCDD-mediatnl rcuctlve oxypn spdn production has k n p r o w to involve the cytmhmme 1'450 IAl and lA2 (I)iliherlc~ CI ul.. 1997). 1'1% Ah nwptor mdiolnl lndwtion of runthine oridnx'nnd xnntllinc Jchydropnnr activity by TCDI) ~IIS ~llro bccn rc- ~I,,CI~I 1,) k n.*p~\~ihk. lor !In' pr~uhlni~~!~ 111'
'lstrr,tb~xr 171 i?lYJ?I 127 /.IS Ill rcuctiw oxygen rwim in liver (Sugihilrs ct ill., 2OUI). In the pracnl aludy adminiutrution ol' TCDI) dccwd tla ttctivitim of ci~t~ilitw, xc~pr. oxide diuilutuw, &IuIathio~ic rcducttt~, itlid 8111- tutliiolw proxiduw. wl~ile the lcvcls of bydrop11 peroxide and lipid peroxidntion incrwscd in tlrc mitochondrial and microrom~l fnctlonr of the testis. Mitochondrial rapintion is the main biological wurm of tuproxide anion radicals undcr physiological conditions. The mtioxidanl enzyme supemaide dimutasc hps bmn rhown to aculerute the dimutation of the superoxide anion into hydrogen peroxide (Nirscn and Knyscl. 1983). Cntulase allows degmdation of hydrogen peroxide into water und oxygcn (Jeulin el al., 1989) and llic glutathione pcroxidue/reductue system ha^ been rhown to calalysc the degradation of hydrogen peroxide and lipid hydroperoxide by using re. dud glutathione (Alvam and Storey. 1989). The reduction in the activitin of antioxidant enzymes and incruuc in hydrogen peroxide and lipid peroxidation could reflect the adverse effect of TCDD on tho untioxidunt system in testis subcellular frnctions. Oxidative stress has been shown to occur primarily in the mitochondria fol\owed by microsomn. Orievau and Lunnou 11097) rcprtnl that ROS such as HIOt uppca,rs to k u key agent in the cytotoxic effects In spemlowe and in udJition to their dim effect on cellular constituent produce oxidative stress by dwrc~~sil~y the c11zymatic JeRm of the testis. Un& high ROS mnrrntrntions pre-dumuged rpcnnutowu an exposed to lipid peroxidatioli by polyunwturuted fatty acids (Ichikawrr ct al.. 1999). The mitochondriul and microsomal membrann have ken reported to undergo permurbility changes following enhanad Lipid peroxidation and Jutathione depletion (Chancc ct al.. 1979). One of the important upatl of anlioxidwt en. rymer ii their nature of synergistic functioning: a decnrsc in superoxide dirmulav clctivity hm been shown to increase the level of supcroride anion, which is known to inactivate catlrlrro llctivily (KOIO and Fridovich. 1982). Similarly. when wtahuc or gluuthbne psmxiduc faib to climinate H,O1 from the all, the accumulated H,O, hru been ahown to caw inactivation of suproxide dismuw (Sine1 md Garber. 1981). Reactive oxygcli spccics Iisr bceci sliown 111 d;~tiii~yc ;llliioal ill1 ~llscromolcculcr. viz. prolein, lipids. ccc.. i111d lllc cliiinpr in llic rcprtnh~ctivc orpiill wciphts 111:ly slso Iw duc lo iIi~~l~ii$c ~IIIIIIIC~~I hy IUIICI~VC Uxygc11 rwiw (&l~~ultdt%rl: IDYO). 'I'llc ~.acllla sugycst thut the low doses or TCDV clicil dcplc. lion in antioxidunt defense system diflerentislly in mitmhondrial and microsonial rrilctions indici~ling TCDD-induced oxidative strcsr ill tculis suhccllular fractions. This may lend to disruption of functional integrity of cell organelles. In conclusion the adverse long. term effects or low doscs or TCDD on male reproduction of mts may be due lo the induction of oxidative stress in testis. Acknowledgments Tlie aulhors ock~~owledge Dr Slcplien Safc. TX. USA, for the generous girt of TCDD and thc still'l' or the Bioinrormutia Center. <strong>Pondicherry</strong> Unlversity. for various incilitics. C.L. acknowledges the Indian Council of Medicnl Rcseilrch. New Delhi. for Senior Research Fellowship. K.C. ilck~iowledges the Lady Tstu Meniorial Trust. Munibai for u Junior Scholursliip ~IIIJ P.I1.M. ucknowledges Ihe Population Council. New Yolk for financial assistance (Grant Nos. B99.047P-91 ICMC md B99.048RIlCMC). Ra. 23. 77-90. Bury. I.A.. Ausl. S.D.. 1976. Lrclo~roridlw cill;tlyml Ilpal proriblion of micrororn~l and ~~rlilicrrl n!nn~brnnn. Biochim. Biophy+ Acts 444. 192-201. Curlbrre. I., Mmnmik. 8.. 1975. Purifimlion und chlmtctcrl. ulion d Ur Ilrvanzym plulrlhiom rrJurlax from nil livcr. J. Bbl. M. 250. Yl4-5480. Chainy, O.B.N.. S.nunwrry. 6.. Salnanla. L. 1991. Tnlov temne-induced sbnp in taticulur nnlioxiJilnl ~~(ICIII Androlopiu 29, 341-MY. Chuncc. 0.. Sie* H.. Bonrit. A,. 1979, llydrapcro~de nlslubolirm in murnmulwn orylnl. PII).riol, Kcv 59. $27- 514. Chw. S.E.. 2WO. Endwrinr disruploi. innd nlalr rrpruducltsc futction - a &an ncuka. tat. 1. Andrul. 21. 45-46.
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FUNCTIONAL STUDIES ON THE EFFECTS O
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ACKNOWLEDGEMENTS I wish to express
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TABLE OF CONTENTS Page No ACKNOWLED
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TABLE OF CONTENTS (Continued) Deter
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TABLE OF CONTENTS (Continued) Effec
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TAULIC OF CON'I'ISN'I'S (C'untinued
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xi LIST OF TABLES (Continued) I'agc
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Table 24a Table 24b Table 24c Table
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Fig. 1 Fig. 2 Effect of TCDD on the
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Fig. 12c Fig. 12d Fig. 13 Fig. 14a
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LIST OF FIGURES (Continued) Fig. 23
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LH M PL mg mRNA N AD NADH NADP NADP
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layers, an outer layer of visceral
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tubular epithelium and continually
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lmn~unization of monkeys against FS
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and this reaction is catalyzed by t
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extratubular environment or synthes
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the most active in protein secretio
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1958). A rapid fall in serum testos
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have also been reported in the rats
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Effect of TCDD on heart muscle has
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1.4 Effect of oxidative stress on m
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2 SCOPE OF THE PRESENT STUDY There
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3 MATERIALS AND METHODS 3.1 Animals
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Subgroup III: Administration of TCD
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choppcd with the help of sharp razo
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anti-FSH antibody. After incubation
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lust sped (500 to 700 rpm) on an or
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3.13.4 Quantitative determinntiun u
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The plates were dried by inverting
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centrifuged at 3,500 g for I5 min o
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3.17 Preparation of tiasue humogena
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supcrnutant was taken. Hlunk was pr
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3.21.4 Assay of glutathione peroxid
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4 RESULTS 4.1 Administration of TCD
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seminiferous tubules was found to b
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fractions of rat testis as compared
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4.1.11.1 Effect of TCDD on the prod
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4.2 Co-administration of TCDD and v
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4.2.8 Effect of co-administration o
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4.2.11.5 Effect of co-administratio
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Table I b. Effect of TCDD on the we
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Table 4. Effect of TCDD on the seru
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Table 7b. Effect of TCDD on the pro
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Table 8c. Effect of TCDD on the lev
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Table 9d. Effect of TCDD on the act
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Table 13. Effect of TCDD on the ant
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Table 15. Effect of TCDD on the lev
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Table 16c. Effcct of TCDD on the an
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Table 18a. Effect of co-administrat
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Table 21. Effect of co-administrati
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Table 24b. Effect of co-administrat
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Table 25c. Effect of co-administrat
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Table 26d. Effect of co-administrat
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Table 30. Effect of co-administrati
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Table 32. Effect of caadrninistrati
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Table 33c. Effcct of co-administrut
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5 DISCUSSION 5.1 Significance of th
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Measurement of weights of the acces
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compartment of the cells (Chainy el
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the reduction in daily sperm produc
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eported in the TCDD-treated rats (M
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lilnction is to ililcrccpt lipid pc
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in the animals administered with TC
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protective effects on serum testost
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suygcstccl that vivamin 1: conld im
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Fig.4~. ENect of TCDD or TCDD and v
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tE W d In w it I I (,OCX) 3s PUB (,
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Fig. 7c. Effect of TCDD or TCDD and
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u 3 f ~ = m w * N O .- ~Slew aew en
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Fig.lOb. ENmt of TCDD or TCDD and v
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Fig. 1Ir Effect of TCDD or TCDD md
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Fig. 14c. Effect of TCDD or TCDD an
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Fig. 16b. Effect of TCDD or TCDD an
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Fig.18~. EKect of TCDD or TCDD and
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1 1.2 l4 Fig. 19b. ENect of TCDD or
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Fig. ZOb. Elkt of TCDD or TCDD and
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Fig. 21. Effect olTCDD or TCDD and
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Page 175 and 176: Fig. 24b. Eflect ofTCDD or TCDD and
Page 177 and 178: 7 REFERENCES Aitken RJ, Irvine DS,
Page 179 and 180: Chitm KC, Latchoumycandanc,C, Mathu
Page 181 and 182: Flsher JS, Turner KJ, Brown D, Shar
Page 183 and 184: Hornsby PJ. Regulation of cytochrom
Page 185 and 186: Latcboumycandane C, Chitra KC, Math
Page 187 and 188: Moore RW, Panona JA, Bookstaff RC,
Page 189 and 190: Rommerts FFG, Cooke BA, Van Der Kem
Page 191 and 192: Slaunwhite WR, Burgett MJ. h virro
Page 193 and 194: 1.1 Preparation of dosing solution
Page 195 and 196: 1.4.6 Wash concentrate One bottle,
Page 197 and 198: 1.7.3 Estradiol controls Two vials.
Page 199 and 200: 1.1 1.2 Orcinol(6%) 6 g of orcinol
Page 201 and 202: 1.16.2 Horseradish peroxidase (8 Un
Page 203 and 204: 1.20 Superoxide dismutase 1.20.1 Tr
Page 205 and 206: 1.24.2 Thiobarbituric acid (TBA; 0.
Page 207 and 208: Chronic effect: of enclosulfan on t
Page 209: enbw1h1.W mu IUY Wcab unpiliml~t at
Page 212 and 213: cbniuis 11~11 b"nci:~lc ~wlive oxyg
Page 214 and 215: DNA, RNA ;uxl Wcin wcrr abarval, wh
Page 216 and 217: Mt~~m(10-lZsu&)vacob- Wfmn Ihehml ~
Page 218 and 219: New Delhi, llrliil Ib SCI~KK Rcm& M
Page 220 and 221: tiii~lc rcprmluction or rut (Cllitr
Page 222 and 223: wciglit (b.wt.) per day for 45 days
Page 226 and 227: Chilr:~. K.C.. L!olnn!myci!~dum. C.
Page 228 and 229: C. hteboumyeradlos . KC. CMtn . P.P
Page 230 and 231: I. 1 Ilal 01 2 1.7 # I c I ~ ~ L I
Page 232 and 233: Fin. L Corrclatwn bclwfcn "perm cou
Page 235: Iionshii nepmdvctivc mxicity ad upt
Page 240 and 241: o. Glutathione reductage mg pmWn mg
Page 242 and 243: IOWAME -. Tor. #558517 PME: 9 SESS:
Page 244 and 245: JOBNAME: Rqm. Tm. M5W7 PAOB: 11 SBS
Page 246 and 247: t 02 ELSEVIER Toricology OM) (2W1)
Page 248 and 249: ---- - wcre obtu~ncd from E-Monk. G
Page 250: 3.2. A,III~AILNI syrlo~t of ,nIroch
Page 253 and 254: Uc,#!dy. SC. N;tdcru. S , 1% Conlrb
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