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?1:e{1ifroxrn{r1ur11dn{ il?J. fld + ariud 2 nrn1.lerru - #uernru 2537
m'r { rfi a ntuai'Lunt € Fr sJ ri'l rfioTo st#Porcine zona pellucida
or3fi r-rrrotni::'oriuo{ frnm"nf
vtU?OOUA
nr:n I rir tff er"Lu#oifi u nr ri fi dr uqi n :ii fi da 6rio rf, u rion Eir.:nTutrj urqiififiq
^d(ctl-tutJlJ 2'lra0 ?DlTtJflfrun:Til UUO:rnr.:noirru:rrluro n:rxnl:yirenun::il orriju* 1 w*A , , a *r -a! o 611 , t d u ed A -o 0J LTzu?1 tltuntl ou:rLrLto rr'r,ro ir-iurrvr moi.Lta lillrn nollril n n'[ri'dn rfr oci'a.:nr:ql s
-.o--o , a i; r !t r V - | X !uonolnuu.luuflflt.ttou.Jtru wqon::rrlnlurrrlolLrJ uruuntrrl{u rdt.:oonr:'l'r.3u1finu rrn:eirtdqirueiaudr.ra.r airuiinr:rir#q{:rfiir{riju#ooiolfirLirrrxaruiu
U
n-ntT.: urn:o'rriiuoia'r'ld:-rnrri.;rar"Lelciornn"rra.rtunr:#.:rnnnr:riiu#oro.:fi'n{ fi.r
rrri'jrf,oq fuorfini:vt-oL u-lnlrel )r nr rfi,ilFr u nr:ff urruiaaaflruuEiifrdfi oro rfi e zu n9qv a g
^d 6l , < <r Arirrrfrr.::r:Jyri.:rure.lirnsi6nr:lii'urffriiuerrrrri,:rrnil:;nrrfi1-r.r4- 1 )^ 3 . ^l ada. u v taveatua d a I n il il ulr I n 01 uar n n r :fl r.J nr ruo Lo 116o r n nr rir.: o'u il o q I u6 I'Loin nrr
^:-,-.^^---ylql]urnrrnelrirtfro Leu"ldi6nr:vrr.:nfi rYrn:uivrurfiu 6a nr:1d' antizona peilucida
antibody tr"l#iunr:nrnrrfio#qiun-,usfiq:run"*uu ifiaufrlruurfurfioornifrnr:erru9u9
^.-.-o ^aJ A ^oX --..tt..oj -jq - d I L dAA q e, d t qvt n,nituel'lDau r.:rruulitjuinnrtvt[fzuno ilnoonu ill6nrtLu';ru ifrueir"["i'siru'l:.ial uns:a q v d uG A -
a
fl at flr tllTn n ntJ&Jtil n n lo o n ul oo o.: nr:nrrru6otunr:triTrur rnnqfioir (zona pelucida) lunr:qrnrrrfio r3ruorn
q
^ J, q ,:_^.e^i^. ^V., ] f , o <t a a a,6- , &n?tilnqm?rlunr::Jfiauluu riusrouufr.lfr.:sirriJu6o nt:f]flrilTilto.tr,.,l-.lur1{ iryur,^..^^.3-v- ---?' ard a T e <tLylnauq'l n1l0lJ1J0.lflt:-lirululirto.: lryilr tnnnfroi'r astiJuoqtiuoi'ulolnr::Jfrauirunro'rriJuqionr:fl.:qirro,:o"'reiau q-.rfuurnfin1:yi.r01unn'[nnr:rfio:.Jfrniur::ra.jr.r
at c & Ta tTJ 5 ru n u lrur n l n n fr oir fr o : ar I r : n fl o.: ri'u n r :rJ fr aui'loi
' ntairtwatfrinn nruz#ntuwnuffisa{ xu1inil't#ulJauuriu {tniarauuriu 40002
? rlnf;nr*r&'qrunmri nr*:fiorunmuersn{ rnrinora'urouuriu o-.rur-o:rouuriil 40002
a9
90
Zona Pellucioa (ze) riJudula.:vat-4,
fl1:no1u[on1qu (gelatin) ry.]aUToU'lut, - (N ehit o.: d'qi 16 u.: nnoi'r uul tiiusr::Jrin ou
qJ
lnoInLJ:6u drfinrualuGtiju tissue-q
specific ttsililtilu species specific firarir
d rfl u specific receptor rilaiulv{ariJillua,a r& e t v ll r
flil56t1t1{ 6l il'tou unlta1i9{'11{t?1 UJT'Ilrd x, a ^!q?nuil'rrnRUfllan t1 rnqn'r:uilfl1151j1*
Tmir rn anfi oir :J*n ousT'r ulnnTnhj:du.U
Ad'3 5Uq ne ZP . ZP . ZP s{ ZP t:Ju
1' 2' 3 3
sperm receptor rafi'ndrirrurlfHfi qindu
nilnh rfiq (contraceptive vaccine)
n1:loligil Porcine zona pellucida
lqifi nr:ilTfl ulnr:tqriu il Porcine
zona pellucida a'rH:-utfrfl uuou6 tail
ranru?f uqrr#nnr:vr-rLjda vrhtvfT v sU, A A :Lor.: flT't.tr 0{T.l [u n n o qfi T 0u 9t til ua ilci'rurfin 1 ufrr6.:rfiubjuog zona pellucida
n1R follicle
n1Tlaiuil Porcine zona pellucida
(Henderson uaiflili, 1 988). uu,i r 6 lle Z <
u't r.l t1J s.t ttT ttlj.l t? tJ't 0 r n1 u u't [tlj.:evXAvA
un?aqtuotuoTau 6J oan uqo?u[nTo.]Xeauotilo uei'rtdr ice-cold saline (O.9Eo)
TaIl6lui{ thomoginize) riruorrrir u'[un ou
llu'ret 5o0 rroi ZOO 'lruIn:rrue: ,duXAAa
rah tfl r*rfr a ril ofr ilr ur olraqj o on u#r n:or
iiruqrdrururo 75 hTqr:rilq: rdo
rfiuciruto.:'lri #rtsirl ice-cold saline
KKU" Vet" J. Vol. + No.z July - December 1ee4
rrd'rriru Percoll gradient rn-.ta1nfi'u d1
tdd'ld'rur ir u n pe rco I I o a n lah 6'luii'lu*[qi'
zona 6s:; rrd:#rteiruil:;no:rttneidu roan yi''t zonaheat-solubilized d t6'n,ZO
aAqdr,1*'t11 t1^t0 Lu [oaluta.: zona pellucida
eglycoprotein a''lnil ilYt't zona
ultracentriluged tt#ri tn:r rvirar rJi lr nrfrAeadaffrAAoLilTOUO? U?! O[An LOT Lil nfl fl il',] fl11.{1 0.]
<VtduAo$,Qezona [nt L? Ll.tflR']?to9rtt14.: tilaul LIJ Lr
CIA Lil
AAd?fin1TilO
o f , ov A ueu'r [sl{'r tl^tflfls9l't ytttun Lo ttnt< lld 9 v
tnu ['] Luflnl?30qttil.iil1nsn1u tta')q,
tlflil9i?U Freund's complete adjuvant tuA , u qqlllva
ilTil10Ty1t141nU !U L9lilTil'lqr 1 ildnnq:
(r-l::lrcx 2000 zonae) dorriT nd'rrarilg n*r Iquvirrfu:Jrsil'rru 1-2 fr'rjqrf
rrsioslf Freund's incomplete adjuvant luAe&,u'-Y
n1Tilq nTr qtilnT.: ql 0 [!u analnnT.] ttTn3
, ll da e o a0 u''t.: lTn q 01Q0!q 0.:il1n1Taa
uYanrrd'usilAnua{aln 3 rfiruu:n 9 r6ou
AAAddquT0nlTqotilu.ttljil to u? [uluttTnl a{:3 ut
proestrus frorovir'[ai' 6'oo'.lnzuan1:r q e s u E u I4v
vl q n 0{}lU?'l t14 9{ nLJ 0.: n iln1:9l .lYl 0.: [91y,yn.: 3 ttilll1Jo.:nr:n:lgtu
n a"[n nr :fl a s fi]$nr :rJ fi ar+BAq
rfr o.:orn.Lunr:rJfr auG qiolfi nr:t at6 u & L Av
0unuT!14'r'1.: fl !:JTil nLTu L{u1 til n nTgl't- Z " ,i , r -t9ltuu141nfllnlTt! nuuttil n.: tflT.l fl:1.11J 0.1
v d a,a - d?153{1T6lsl1rry{nUFl'ldn5 il?J. ilyi 4 ouuYr 2 n:ngrelru - iuererr 2s3T 91
Iryr.ir ir^rnnfiai'r fio:arrur:nil0.:riunr:qt
ard Vl e v Ltnl31J0.lfrtilTu [o nlTnT;ou9l1u porcine
zona pellucida orvrrld'ir.:nrrair.:Joqvazona antibody lJ urJ'r 11t [u tn o
vimmunoprecipitation uuT1.[u0nt0.] zona
pe|Iucida fi zu n r or-.ru3on-oIrr.r qir rruu.rdAd a a <, V! A o q tflLilTilA:iltA! 0.1t1lunn Inilil't Lun]5
Ju,u-ililflufrno1Ju Lil [o
619/ 5l v drunZo0nlT[5 zona pellucida [Ua{Ot
n?'l,r Turner !!n:fln{: ( f OOZ) toi:ru.:ru
e.r fi I a { nr rtd'io 6un ru rir rfi olun I r lo'r ur uI
d vl ,o t o o J q/ ol 2nfl 0 LN:.Jnl:o.tyt0.ltno:i]uLun?liY--vt,La
il.t 7 a't ttn: Llyluzundl.ltnu.:
nrvsilu Bhatnager L[n:flo{: ( i OOZ)6)q
:'l U J lunlT tT deglycosyiated zona
peilucida tunr::l o.:funr:ilfr au81u
n::qir ri r^rriir'loizunvrn d'r uqifi zu ni!n?iln r :n'r J'r 1.r1 o.: i.: iri Il u o u -, r* n r : e ;r.r td
avj r Z - ovl , grioilnq Lil il't14un?J8nT.t'L?ilom.t T?lJli.1
" vt ,i011{?14 t"}l?1fin
Jones Lrniotru; (tsoz) Ttunrlr'd1't tJi{afloatu"tuta{ primary, secondary,
tertiary follicle Iou'[lriolvfrfionr:'j uu, - & a
til n uiluil n.t?t''1.:o nil u'tr? o.:i.t'[d e.:uuilq
ariunrlunr:"[ri' degtycosytated zonac
pellucida rfl u urouGiourdor^ieiurindu,u,
n3.Jn1tuo9,t0lil
#r Kirkpatrick !!a3nru3 (1992) 11u.t1u
nrrld' porcine zona pettuciOa (eZe) luvtal 9
nlTO8JnltilOrJ't?'t n1T[14 PZP OqCIonuq
...df(e3 il nouno9.lflil$rur 6[]il1Tnil0.:nunlT9
&euv,oi-.:yio.r'[oi'lrnnr'] 9oo/o usr arolruil3rur rxnr:n ff.: r o sTo :ro il0 o n.rtuflu'r.r o--l
Liu rrnrnil: ( t sa0) Ttu.tlunlTqo & u e, tL5 PZP 4 flTn ill,rnu 2-4 flilo-rl4 rrn?
u Y a !.q a Anriouslann;nlutgrOuyr 6- 1 0 lrd{aln
o Y qe Y - q on'']Toq nri ttrn Lu ilail 0.t nunlTo,tyl 0{ Luil1
86o/o L[nsr;q;iln fi ri'rriut-.1srrfi orifi ,: 7gcs
rd ou Io utrinrorr l fi orJn 6r o.r olnr:qrfae
::oltfl aT [ilu u:afl nl*ilsrtJanliu']f6s Sacco LLniFr{ur ( 1 9S7) rru.nunrr"ld'
zP 3 al n fl n:tufi.:'ir ar rr:n :J o.: ri'unr:q
Y - u' T9tJ?18,1 tO t9lU81A1\!nl:T!niUnlT
o o\4 ' u I,?'1 1,:''r 1.tfl 0,1 TJ LlJ t[d :'lr l_J i u n'] : fl T''].1 LlJ tto
- Y a o B ,cufl.3alnuil 10- 15 Lool-r ;.1lflnfl]illTno iu-
n tr rryiru',irfr'loiqr ilil nG
qfio Shivers lreinrut ( t Oat ) :ru.:'ru
nr:hi' pzP'Lusu-r'ir au-lfito"ir nr:99
n::ci'uo"r u PZP a;fl''lrirrnfl orri''unr:Y - uv ro.:yl0J [o Lo ua:wL immunoprecipitate
^ 4 da
formation tnq?u?lil-lo'tuu0nuan zonaA
pellucicla fi,:nro'jr immune complex fi^ A d n,v atnol u el 9Jn t:JlJo??''1.:niauqu.ta otnl3A u arr' u U 'uT0 nt::?tJ 911J 0,: fl tlJ:il nil L1
Mahi-Brown llnrnrut (1985)
nu'j1 adjuvant fl1ilt;nd?un:sei'u:sot-uq
rl6quA-YafLota 0: t14fl.t?t anasuu uinlTrufluoto [o u,u
. Vt ,, q ea e ^ iaalum 0r [iln0 tntnont:[ttltu:t'tilytaq ttqi
astli'loroof'[rin.:rvir Freund's rrns cp-g
20,96 1 adjuvant airunr:nJiuurfi uuntr99
nT;o! uft? u crude porcine zonaeq
92 KKU. Vet. J. Vol. I No.z July - Decembeltgg4
^EN,.lriavrifiu rriu nrrlri' Purified Zp3 alphaq
rata Zp3 beta-oG tun:sqi'rulou'[rjyi'rq ea ,l uvl,Lu t n el nl T til n u u $il a.t il't.l 0 0 1,\ u't T?1'1.: T.t L?
(Jone un;nil!, 1 992) uonalnd Keenan
Lrnrnrus (t OOI ) a-.lwu'jrn'rrrrunrrou6reuquaSJu,ooLfiuTflylt 1{1.[ A; Lilvl't Lfin1Tyl1.1'11.[
& Vl ,a , all r o q d elJ0.tT.J [1J r,r OilnO Lilttnst[1{;U'r 114 [T
Deglycosylated Zona pellucida rai o
zp3 a'[nnInI:J:6utfluuau6routunr:^ualqduauRqlnflunlJnltuo fi'l Lunl:tgr:uil?n{u
I
qrruni rfioluourn q orovir'loilo unr:#r r o :r s fr-n 6 uo r n r rl :J'[ q qf,ti'r r o rr r f
AAfUfi T 0 n1:LqT U il LO U 0',1 fl U tYt fl Ufiyt't.t1 t ut?ern::r lunr:uun DNA a'rn ltr.ir rnn
n8qi'r ttnio'rflu Vaccinia Virus tijunru;qt
q q eu a o a[un1: Lfi 1 Fl{iln iln'] 11.t9 9l.t t5u? 9lsil
Ia q i.lv 'q v
1rfl'ru11.{o Lilnuil txau tul_Jo0:Ju
nlTnririltfioTo uld Porcine ZonaI
Pellucida riutijuif;drJn oon-u virtoiir ur 4 w' Vl ' v d v anl LTA-] U Lil fl ,i rLA! fl 01 fl',] tJ',];nn nu il''lil 0nq., g
vlrd ! - ^e v a t a
tq 0 n til 0T3q! niln Nnuflo 0.t n.t ttlt0stJ{q
:l ru u r t u I r { n r rfi ei o n'r r yf 't .r'r ill o .: :-.: trirra:::qi':taoillu rfi o.:ornu-.:o u'1u
9l
Tr fi 'i 1.1 n 1 :6 n rsr n r :1 {"8r l{t n 1 :d'l'lilld'fqAovd
nl q11 !U0U't fl qa3 il nlTul? nSilfl ilnt tugl
Iq u Porcine Zona Pellucida trtri'niu
adr.:rtniunru
pellucidae (cPZP) n-upurified zona lrac-
tion
(peze) ilui'r n.rrnrreiuei?u cPZP a!{v ll cLnrun [otoo5fl.]uns:J1una1.: ttdsol0 Lno
.il
a <r v.l An?lil iloilnqi I 0.:?{TaunlTLljl{do tunr{u
fitorrqois.r ei'runr:n:;{uoiru pPZP o:
lra*lq ror oi::o-u:Jrun nr.:'].rra un'r:tiJ us&
floilnqt lqun.J CPZP un; pPZP fl'ril]:na Uv
tJ0.:nun1T9t6lilqq t9t
riodzaon,r:l# Isr.irOvdtr,[6{A')
rnaaf Eir
t ) fi onrurJnaqn-uun!fl;q?n. aa q ev a o l! w'
2 ) ?5',n']r 114?nTuYn [9t.]'ru
s) rirlfdru'[r.ia.:
+) fi nrrrudr til'rrei arruufi::n-ufq
fUrd,daAtq u tilil9{nq 0fl::l11 u'r01.{ 6l
5) nrrr:n n d''ururfi p n'[ai'6n rfr o3,t
Iq0{nt;
,t A O !f/l , ^ IuatGluzaf fl1Tt5 tsl{1 rilnafio1
$e a Ur, a
01aytuiln11.:tn u.: Lo lu]J1.:nrileaiveaa
zunll1.tLfi u.:Yt Ttu [o n 0 il un:iln?un1To uU ' o q e p !l ,
Yt'l.l'l u? a.: T.t LlJ 1,1't Lvt 1 u ? unl T fl :'1.1 Lll
fi qiln6 siruru'ld nn unrrseiilrosTorrxu
n9nnt u da v a
0u'1.: [Tnq 10tfiuo.]na1? 01a
ruriltlai'lq unr:ld'uauG reufi fi nr:yirtvi
?1:fl1rfi'o?ulrnsc{1aor{ N". fld + ariud 2 nrnglrrrJ - #r.lernru 2sgl
Rxllind 1eilnu1rr. 2537. nr:virrariu#o{ Tr,:firv{lrariyrur#ur::ilernoi nr.lrlrlte
ZqS uthAitken, R.J. , Paterson, M. and Koothan, P.T. 1993. contraceptive vaccines. Br. Med.
Bull. 49 (1) : BB-99
Bhatnagar, P. , Gupta , S.K. and Sehgal, S. 1 992 . lmmunogenicity of deglycosytated
zona Pellucida antigens and their inhibitory effects on fertility in rabbits.
lnt. J. Fertil. 37 (1) : 53-63
D'Occhio,M.J. 1 993. lmmunological Suppression of reproductive Functions in male
and female mammals. Animal reproductive Science. 33 : 345-372Gupta, S.K., Bagavant, H., Chadha, K. , Gupta , M., Yurewicz, E.C. and Sacco, A.G.
'1993. Mapping of lmmunogenic Domains on Porcine Zona - pellucida
3 - Alpha - Glycoproteins and Beta - Glycoproteins by Murine
Monoclonal - Antibodies. American journal of reproductive lmmunology.
30 : 95-100
Henderson, c.J., Hulme, M.J. and Ailken, R.J. 1988. contraceptive potential of
antibodies to the zona pellucida. J. Reprod. Fert. 83 :325-343Jones, G,R., Sacco, A.G., Subramanian, M.G., Kruger, M., Zhang,S., yurewicz, E.C.
and Moghissi K.S. 1992 Histology of ovaries of female rabbits
immunized with deglycosylated Zona pellucida macromolecules of pigs.
J. Reprod. Fertile. 95(2) : 51 3-25Keenan, J.A., sacco, A.G., subramanian, M.G., Kruger, M., yurewicz, E.c. and Mogh,
K.s. 1991. Endocrine response in Rabbits lmmunized with Native versus
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effects of porcine zonae pellucidae immunocontraception on ovarian
function in feral horses (Equus caballus ). J. Reprod. Fert. 94 : 437-444Lir,l.K.M., Bernoco, M. and Feldman ,M. l gBg. contraception in mares
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94 KKU. Vet" J. Vol. 4 No.2 July * December 1994
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Progesterone Levels in Estrus Cycles. Biology ol Reproduction. 32 :
7 61 *772
Mahibrown, C.A., Mcguinness, R.P. and Moran, F. 1992. The cellular immune -
response to immunization with Zona Pellucida Antigens. Journal of
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