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Phospholipid analysis in testis-epididymis complex after alpha-chlorohydrin administration
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p h a ...h ..l o r o h y d , o~5 ~ g dy ght ...for . 14 days, did no t t er to t pho sph o i i pi d s, ph o s~h a tld yl c ho I i n e, ph o sph a ti 0 yl eth ola ne, pho s p h a t i d y l set e, p h 0 s p h a t l d y l i n o s l t o l d sphin m y e l i ~ in rat t e s t i s d ~aput and epldld s. However m ked in totel p h o s p h o l i p i d s chollne, p h o s p h a t i d y i eth ol e and phosphatldyi inosltoi in da epid is d vas e fe tens ~s " o b s e r v e d , p3a also s h o w e d s i m i l a r e h g e s in p h o s p h o l i p ! d s .
Accepted
for
publication
J U N E 1 9 7 8 VOLo 17 NO. 6
April
i2,
1979
5:23
NTRACE
N iN
CT
ndings emerging from current literature indicate at a l p h a - c h l o r o h y d r i ~ , a reale ....t l f e r t i l l ty a g e n t , h as at I e a s t two s i t e s o f a c t i o n i n the m a l e rat.- at a h i g h s i n g l e d o s e the d r u g p r o d u c e s a c h a c r e ..... s t i c l e s i o n i n the c a p u t e p i did s l e a d l n E to t h e o c c l u s i o n o f t h e al d u c t a n d 6ubsequent tion of ge inal epithelium causlnN a perm ent at low the d r u g a functional i n the s p e r m a t o z o a c si~g 8teriilty. & tuber of s u g g e s t i o n s , b a s e d o n e x p e r i m e n t a l d t h e o r e t i c a l d a t a , h a v e b e e n m a d e i n t h e p a s t few y e a r s to the ~ e c h sm of drug actlon (1-8). A s yet, t h e mech ereb~ low dose6 of aiph c h l o r o h y d r i ~ e x e r t .... tl effect i~ e rats is not cle . obab!y reasonable h othesls has been t e d by "o ....l a (9), o sug s t e d t h a t b e c a u s e of s i m i l a r i t y to g l y c e r o l , ....p h a - c h l o .... h y d r l n act as ant onlst s thesis d met iI . I f it ia 80 tally, ph chiorohydrln Bhould ter e phospholipid c o n c e n t r a t i o n i n the i B - v a s ...d.e f e r e n s c o m p l e x ei e~ qu itatlvely r ...qu . .... e present w e r e m a d e to t e s t t h i s ~ 0 ests. D
ts w e i g h i n g 1 8 0 .....0 were given d ! y o r a l dose, 6.5 body aiph chlorohydr in i ml of 0 . 9 % S e r i4 days. control im s received i ~i of 0.9% s e. e im s we e 8acriflced hrs after the last se. e testis, epidld is d va6 were re weiEhed i e l!pld8 were extracted f ~ epldld is (ca t, c o r p u s c d vas d e f erens £or m ~*m e th ano (2:1 V/V) by e me tho d of ich e t ). Mar ttl' s (i!) ~ e t h o d used r e estimat n o£ total spholi~ids. Different w e r e sep at ed b y t h i n I er c h r o m a t o g r a p h y r th e l f i d e n t ficatio~ and tiflcation. Each isolated phospholipid s m e a s u r e d b y the m e od of M e radio-isotope (p3E) obt .... as carrier-free sodium orthophosphate ( a a Ato C Reae Centre, ). e 18otope 8 tr erlto~e i~ 4 hours e exper e~tal d control m s at a of uCi 0 wt. dete ine of p3R into oration tot , a o a ~ o u n t of l l p l d s e x t r a c t e d is d 6 ~eferens t e n on pl chefs and c o u n t s w e r e made t h t h e G.M. tube c o u n t e r . Similarly, p~R incorporation to rious phospholipid fractlon8 calcua~ter scrap rlous spotG f~~ C plates.
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No slgnlfic t ch g e s in to tal p h o s p h o l l p i d s , p h o s p h a t l d y l e h o l l n e , p h o s p h a t i d y l eth ol e, serlne, p h o s phatidyl inositoi d s p h i n g o m y e l l n in testis, a n d or corpus epidid s as eomp ed to c o n t r o l v a l u e 8 observed ( b l e I). p32 incorporation showed similar changes these ~ol e s. A mark dee r e a s e In to tal ph o s p h o I i p l d s pho sp h a t l d y i choline, ethanol Ine and phosphatldyl i n o s i t o l in c ~ u d a e p l d l d y s a n d vas was ( e I) were II). serine owed a m a r g i n a l d e c r e a s e e r e a s s p h i n g o .... ~ y e l i n i n c r e a s e d s l i g h t l y in C da e p i d l d i8 a n d vas d e f e r e n s (Ta e I); P 3 2 i n c o r p o r a t i o n s t u d i e s also s h o w e d s i m i l a r results (Table Lysolecithin, phosphatldic acid and lysophosphatldyl eth oi n e co d not be d e t e c t e d in suffic i e n t q u a n t i t i e s so a8 have their qu titatlve estimation. DISCUSS
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0 h e o f the i m p o r t t f u n c t i o n s of the e p i d ! d s is to pro v i d e ....en ent r spe ma turation, en spe atozoa pass through different segments of epidid is, a c h e m i c a l m ieu is ma tailed by the e p i d i d is for the ~ o r p h o l o g i c a l and b i o c h e cal ch g e s i n the s p e r m a t o z o a . Hormones, electroly t es, rlo us en z es a o en z e s, ph o spho l l p i d s ....d m y c o m p o n e n t s o f un own origin d s i g n i f i c a n c e c o n t r i b u t e to thi s eh e m l c al en v l r o n ~ e n t o f the e p l d l d Is (12). t era tion s in a n y of the c o n s t i t u e n t s c o u l d i i b i t spe T motility, r e s p i r a tlon, gl yc o i ysl s d m a tu ra tlon vi s-a vi s f e r ti I i zl ng pot e n t i a l i t i e s o f sp e r m a t o a. O n s e t of s t e r i l i t y in m a l e rats, i.e. w i t h l n 48 h r s a f t e r f i r s t d o s e of d r u g a d m i n i s t r a t i o n (i3), g i v e s indic a t i o n t h a t s p e r m a t o z o a in the d i s t a l s e g m e n t e affected by a - c h 1 o rob yd rln. i s o bs e r t io n i s f u r th e r su p p o r t e d by the f a c t th at ...a.l p h a - c h l o r o h y d r i n h a s ...~.a x i m u m ae c lation in the e a u d a e p i d i d y s t h l n 48 h r s (!4), t h u s p r o v i d i n g ma u m op r t u n l t y to a l p h a .... c.h i o r o h y d r i ~ f o r c h e m i c a l i n t e r ac tlon, i.e. ch 1 oroh yd t i n ac tin g as a ph o s ph o l i p i d an t ago nl s t. it w o u l d be r e a s o n a b l e to b e l i e v e that ...ch . ge8 in phospholipids, both q u tltative d qu i t a t l v e , m a y c o n t r i b u t e in the i n d u c tlon o f i n f e r t i l i t y by w a y o £ a l t e r i n g the m a c r o m o l e c u l a r ore i z a t l o ~ of s p e r m corn n e n t s d / o r ~ e d i a in ieh sperm mo sentlaily, a phosphollpid molecule embodies a glycerol (or y other cohol) f a t t y a c i d (e.g. o l e l c a c i d d p tle a c i d ) p h o s p h o r i c acid d base such a s e h o i l n e or eth 1 he. Interference i~ y o f these ietles leads to a b n o r m a l p h o s p h o l l p i d ~e boiism.
J UNE 1978 VOL. 17 NO. 6
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3751z 142 902~ 110
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6391~ 290 361% 176 917+_ IC~3 78~ 80 930+_ i35
7545~ ~18 4 213 67~ 90 772+_ 95 1 !42
7470~ 212 40~ 120 66% 79 705! 72 1375~ 11t
699~ 211 387% ~35 6~.,+ 70 700+_ 85 t400+_ 150
3920+_ 3~ 175% IC~
521% 314 372% 28~ 2605~ 140 1!7 125 67~ 76~ 75% 70 65 3i 50 59~ 5g~ 257~ 45~ 104 39 26 40 1425~ 1 4 4 % 167-5& 15t5~ 210 66 105 57
6925+_ 581~ 180 360
* gicrograa of phospholipi•/gmof tissue weight. ~t~ndard deviation of the me~
Phospholipid Fractions: ~hospha¢i~l choline Phosphatidyl eth~ol~ne Phosphatidyl serlne Phosphati~yl inositol Sphingo~el~n
Control
~p.
i45 14!% 118 5}~_ 42 !4% 37 174~ 75
Changes in total phos~holipid and various ~hss~hospholipid fractions of testis, epidldy~ and vas deferens after ~phs-chlorohydrin ~istration
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osphatidyl choline d phosphatidyl e ol ne have been re rted to be involved in the d r e s p l r a t i o ~ O f sper tozoa b i n o s l t o l has a stimulatory On mO t i l l t y p and E l > c o l y s i s o sper~ato a (I~)Decrease of choline, phosp h a t i d y i eth el i~e d phosphatidyl o s l t o l in the C da s d vas d e f e r e n s in the p r e s e n t I n v e s t i N a t l o n le the ...d.e e r s e motility (9)d respi ra on (16)in cauda is vas A decrease phosphatldyl choline in rat epidld is a f t e r d r U g a d m i n i s t r a t i o n h a s also been by M o r r i s . (i?). observatlons conclusively sugNest at d e c r e a s e in the moie ..... tle8 i6 r e l a t e d to e c r e a s e in s~e me t i l l .
e c o n c e n t r a t l o n o f glycine, actlve p r e c u r s o r of eth el e r plasma increases a~ter chlo hydr treatment ( 1 6 ) S i n c e ...eth . .....o l a m i ~ e is a base component o f eth famine i n c r e a s e in N! Ii lead i n c r e a s e in eth el e and a d e c r e a s e phosphat y! e i i e. A decrea In pho atlayl ....i d phosNatidyl e el n e in c da epidid is d d e f e r e n s a f t e r t r e a t m e n t m a y be p a r t l y r e s p o n s i b l e for i b i t i o n of m o t i l i t y d since these have been I m p i l c a t e d in spe meta l i 6 m (i2). A d e c r e a s e in p h o s p h a t i d y l i tel theels h a s 80 been r e p o r t e d in e c d a epldld is a f t e r drug a d m l n i ~ t r a t l o n
().
A p e r t i n e n t qu e s t l o n ich arl s e s ! 8 h e w a l ph ....ch I o r o hyd !blt~ pld b l o s y n t h e a l e s p e c l f l c i y in the c d a epldid Is. i n f e r t i l i t y ~ hrs after drus admlnist %ion, m a m ace u l a t i o n of the d r u a in the cauda epldid Is a f t e r ~8 h r s d s u b s e q u e n t ch Ees in the c a u d a p h o s p h o l i ~ i d s 6u est that the d ~ N i n t e r f e r e s in phosphollpld i n the c da s an~ deferene, in p h o s p h o l i p i d g l y c e r o l en the ta ilc p a y r e ! l o w e d by its p h o s p h o r y l a t l o n . sslbl$ the ~ glycerol in the testls e p l d i d is c o m p l e x a f t e r c h l o r o h y d r treatment goes in a n o r m y b e c a u s e the g l y c e r o l l e v e l r e m a i n s al (19). it is quite to b e l i e v e the h y thesls h ~. (5) that ph c h l o r o h y d r l n Nets phosphorylated %hi8 , DL l - c h l o r o l-dee phosphate, a c t s as a i i tot Of phosphate because of I t s t al B il ty to the no al s u b s t r a t e of the enz e, 5-phosphate.
Implicitly, these ch ges y be due t o m e c h a n i s m suggested h ri e a l. (~)in pho spho I p i d 8 and ener g era ring systems the c d a e p l d l d d vas after drug
528
JUNE 1978
V O L . 17 N O . 6
CONTRACE
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m i n i s t r a t i o n could alter the c o m p o s i t i o n of the l u m i n a l vesl cl e or i air th elf for mation ( c da epidld al fluid cont s n u m e r o u s lu hal v e s i c l e s o to m nta e i n t e g r i t y of s p e r m p l a s m a m e m b r ( 1 2 ) ) i n f l u e n c i n g the physlco-chemlcal c h of the s p e r m p l a s m a m e m b r a n e d fertilIz pots i a i l t i e s 0£ s p e r m a t o z o a as a consequence. a l y s i s of these luminal v e s l c i e 8 from c da epldid is d Va8 clefstems d spe plasma m e m b r e after c h l o r o h y d r l n treatmen needs rther vest s.
e thors are th ful to Prof. B . R . a, Head, Depar tmen t o f oph slc s, Pan jab U n i v ty dig p fo r p d i n g l a b o r a t o r y faciiitles. Critical s u g g e s t i o n s from . L L o r i a u x (NIH) . T. n are E r a t e f u l l y ac owledged.
I.
icsso~,
R.J. 5897: A p o s t - t e s t i c u l a r tifertiilty rst nual M e e t l n g of the Boc. for the S t u d y of Reprod. d e r b i i t Univ. Nashville, Tenn, 1968.
2.
Ericsson, R.J. and er V.F. Male anti f e r t i l i t y comunds o Io sic a I pro p e r tl e s o f U - 5 8 97 U-15 646. J. Reprod. Fert. 2 1 : 2 6 7-2 73, i970.
3.
Gunn, S.A. d Gould, T. C Speci ficlty o f the v a s c u l a r system of the male r e p r o d u c t i v e tract. J. Reprod. Fert Suppl. i0: 75-95, 1969.
~.
Jones, A.R., Davies, P., Edwards, K. a n d Jackson, H. t l f e r t i l l t y effects d m e t a b o l i s m of a l p h a d in e rat. N a t u r e 224: 83, 1969.
5-
Mob H., S u t e r D.A.Io, o odin , P.D.C., its, and Ridley, D.D. I d e n t i f i c a t i o n of the ochemlcal p r o d u c e d by a l p h a - c h l o r o h y d r i n in s ~ e r m a t o z o a . Nature 75 77,
6.
S ojlik, E. and 3-ch 1 o ro-I, 2-p ro 2:
7-
8.
,
I.G.
ang, M.C. t i f e r t i l i t y a c t i v i t y of n e dl o i on mal e rat s. o i. R e pro d.
1970.
Vi ckery, B.H., ickson, G. I. and nne t t, J.P. Mech ism of t i f e r t i l i t y action of low doses of p h a - c h l o r o h driny in t h e male rat. J. Reprod. Fert. 38: i-iO, 1974. glmayr,
J.K.
Al
a-ch lorohyd rln induc e d ch an~e s in m - i n o a l t o l and PG a, and
the d i s t r i b u t i o n of f r e e
JUNE 1978 VOL. 17 NO~ 6
5
S
s o f phosphatld
the
S. 9-
p Life .
oi.
~ep
la, J . A . Sei. 8 :
d. ...ii .
ositol the r a t 600, 1974.
593
irago n a d 1969.
real e
t i f e r ill i ty
en t.
48,
I c h , j . , L e e s , M. d ~Io me tho d for e isol at n and
509,
epldid
from 1957.
tlssue,
St lex, G.H. A sl e p u r ! f l c a t i o ~ o f to t j. . . . . oi era. 2 2 6 497-
II.
M a r i ± e t t i , G.V. romato~raphlc sep cation d ysis o f phosphatldes. 3: 1-15, 1962.
12.
g l m a ~ r , d .K. Meta l i c ch ges In sper~ato a durlng epidid al tr si tion: H a n d b o o k o f (e d. Gtr e e .b , R . O O A s t w o o d ~, E . B . ) e eric ysioi. Soc e in~to~, D.O. 437-449, 1975.
13.
rner,
M.A.
fertillty
the rat. 14.
Effects
£ spermato
J.
d
of
ation, IdentlflJ. p i d ..... Re ... s.
alpha a of
Fert.
h!orohydrin u n the c da e p i d l d is o 2 4 : 2 6 7 - 2 69, 1 9 7 1 the
Crabo, B. d De i gr en, L . A . aipha- chlorohyd n mice 30: 1 6 1 - 1 6 3 , 1 9 7 2
d
s trl but io n o f i 4 Crats. j. Fert.
15
L a r d y , H A. and i i ! i p s , P.H. e interreiation of o dative d giycolytl e s s e 8 as s o u r c e o f e n e r g y for bull sp .e r m a. t o z o a.. .. . y s l o l , i"' '-33~ 60~ ...6!3, .. 1941.
16.
O o d m ...., P . D . C. d it e, .Q. S t u d ies on th e . J male t i f e r t i l i t y d r u g 3 - c .......... 1, 2- ..... R e p r o d . F e r t . 32: 337, 1973.
!7.
M o r r i s , S. , D in .... n n e t t , A., o odm , P. a n d .... ~h ..... l o r ' oh~ ....... ' .......... .....t.~,,e, I.G. E f f e c t of a i..pha--c drin the I n c o r of tO the p h o s p h o l i p l d s and g l y c e phospho r y l c h o l l n e o £ rat e p l d i d is. P r o . ......s t. o e~. C . 8: 69, 1 9 7 6 .
18.
S t e t t e n , D.W. o l o g i c a l r e l a t i o n s h i p s of c h o l i n e e t h a n o l ine r e l a t e d corn u n d s . j. ol. Chem. 1 : ! 43 .... 1 ~ 1941
19.
H o d g e n , G.D. e ~ale rat
~3 0
tal glycerol J. R e p r o d .
in rt
the
excurrent : 27'7:2
JUNE
ducts ,
1978
of .
VOL.
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B.S
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UNI
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p h a ...h ..l o r o h y d , o~5 ~ g dy ght ...for . 14 days, did no t t er to t pho sph o i i pi d s, ph o s~h a tld yl c ho I i n e, ph o sph a ti 0 yl eth ola ne, pho s p h a t i d y l set e, p h 0 s p h a t l d y l i n o s l t o l d sphin m y e l i ~ in rat t e s t i s d ~aput and epldld s. However m ked in totel p h o s p h o l i p i d s chollne, p h o s p h a t i d y i eth ol e and phosphatldyi inosltoi in da epid is d vas e fe tens ~s " o b s e r v e d , p3a also s h o w e d s i m i l a r e h g e s in p h o s p h o l i p ! d s .
Accepted
for
publication
J U N E 1 9 7 8 VOLo 17 NO. 6
April
i2,
1979
5:23
NTRACE
N iN
CT
ndings emerging from current literature indicate at a l p h a - c h l o r o h y d r i ~ , a reale ....t l f e r t i l l ty a g e n t , h as at I e a s t two s i t e s o f a c t i o n i n the m a l e rat.- at a h i g h s i n g l e d o s e the d r u g p r o d u c e s a c h a c r e ..... s t i c l e s i o n i n the c a p u t e p i did s l e a d l n E to t h e o c c l u s i o n o f t h e al d u c t a n d 6ubsequent tion of ge inal epithelium causlnN a perm ent at low the d r u g a functional i n the s p e r m a t o z o a c si~g 8teriilty. & tuber of s u g g e s t i o n s , b a s e d o n e x p e r i m e n t a l d t h e o r e t i c a l d a t a , h a v e b e e n m a d e i n t h e p a s t few y e a r s to the ~ e c h sm of drug actlon (1-8). A s yet, t h e mech ereb~ low dose6 of aiph c h l o r o h y d r i ~ e x e r t .... tl effect i~ e rats is not cle . obab!y reasonable h othesls has been t e d by "o ....l a (9), o sug s t e d t h a t b e c a u s e of s i m i l a r i t y to g l y c e r o l , ....p h a - c h l o .... h y d r l n act as ant onlst s thesis d met iI . I f it ia 80 tally, ph chiorohydrln Bhould ter e phospholipid c o n c e n t r a t i o n i n the i B - v a s ...d.e f e r e n s c o m p l e x ei e~ qu itatlvely r ...qu . .... e present w e r e m a d e to t e s t t h i s ~ 0 ests. D
ts w e i g h i n g 1 8 0 .....0 were given d ! y o r a l dose, 6.5 body aiph chlorohydr in i ml of 0 . 9 % S e r i4 days. control im s received i ~i of 0.9% s e. e im s we e 8acriflced hrs after the last se. e testis, epidld is d va6 were re weiEhed i e l!pld8 were extracted f ~ epldld is (ca t, c o r p u s c d vas d e f erens £or m ~*m e th ano (2:1 V/V) by e me tho d of ich e t ). Mar ttl' s (i!) ~ e t h o d used r e estimat n o£ total spholi~ids. Different w e r e sep at ed b y t h i n I er c h r o m a t o g r a p h y r th e l f i d e n t ficatio~ and tiflcation. Each isolated phospholipid s m e a s u r e d b y the m e od of M e radio-isotope (p3E) obt .... as carrier-free sodium orthophosphate ( a a Ato C Reae Centre, ). e 18otope 8 tr erlto~e i~ 4 hours e exper e~tal d control m s at a of uCi 0 wt. dete ine of p3R into oration tot , a o a ~ o u n t of l l p l d s e x t r a c t e d is d 6 ~eferens t e n on pl chefs and c o u n t s w e r e made t h t h e G.M. tube c o u n t e r . Similarly, p~R incorporation to rious phospholipid fractlon8 calcua~ter scrap rlous spotG f~~ C plates.
524
JUNE
1978
VOL.
17 NO. 6
TRACE
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N
TIONS
No slgnlfic t ch g e s in to tal p h o s p h o l l p i d s , p h o s p h a t l d y l e h o l l n e , p h o s p h a t i d y l eth ol e, serlne, p h o s phatidyl inositoi d s p h i n g o m y e l l n in testis, a n d or corpus epidid s as eomp ed to c o n t r o l v a l u e 8 observed ( b l e I). p32 incorporation showed similar changes these ~ol e s. A mark dee r e a s e In to tal ph o s p h o I i p l d s pho sp h a t l d y i choline, ethanol Ine and phosphatldyl i n o s i t o l in c ~ u d a e p l d l d y s a n d vas was ( e I) were II). serine owed a m a r g i n a l d e c r e a s e e r e a s s p h i n g o .... ~ y e l i n i n c r e a s e d s l i g h t l y in C da e p i d l d i8 a n d vas d e f e r e n s (Ta e I); P 3 2 i n c o r p o r a t i o n s t u d i e s also s h o w e d s i m i l a r results (Table Lysolecithin, phosphatldic acid and lysophosphatldyl eth oi n e co d not be d e t e c t e d in suffic i e n t q u a n t i t i e s so a8 have their qu titatlve estimation. DISCUSS
N
0 h e o f the i m p o r t t f u n c t i o n s of the e p i d ! d s is to pro v i d e ....en ent r spe ma turation, en spe atozoa pass through different segments of epidid is, a c h e m i c a l m ieu is ma tailed by the e p i d i d is for the ~ o r p h o l o g i c a l and b i o c h e cal ch g e s i n the s p e r m a t o z o a . Hormones, electroly t es, rlo us en z es a o en z e s, ph o spho l l p i d s ....d m y c o m p o n e n t s o f un own origin d s i g n i f i c a n c e c o n t r i b u t e to thi s eh e m l c al en v l r o n ~ e n t o f the e p l d l d Is (12). t era tion s in a n y of the c o n s t i t u e n t s c o u l d i i b i t spe T motility, r e s p i r a tlon, gl yc o i ysl s d m a tu ra tlon vi s-a vi s f e r ti I i zl ng pot e n t i a l i t i e s o f sp e r m a t o a. O n s e t of s t e r i l i t y in m a l e rats, i.e. w i t h l n 48 h r s a f t e r f i r s t d o s e of d r u g a d m i n i s t r a t i o n (i3), g i v e s indic a t i o n t h a t s p e r m a t o z o a in the d i s t a l s e g m e n t e affected by a - c h 1 o rob yd rln. i s o bs e r t io n i s f u r th e r su p p o r t e d by the f a c t th at ...a.l p h a - c h l o r o h y d r i n h a s ...~.a x i m u m ae c lation in the e a u d a e p i d i d y s t h l n 48 h r s (!4), t h u s p r o v i d i n g ma u m op r t u n l t y to a l p h a .... c.h i o r o h y d r i ~ f o r c h e m i c a l i n t e r ac tlon, i.e. ch 1 oroh yd t i n ac tin g as a ph o s ph o l i p i d an t ago nl s t. it w o u l d be r e a s o n a b l e to b e l i e v e that ...ch . ge8 in phospholipids, both q u tltative d qu i t a t l v e , m a y c o n t r i b u t e in the i n d u c tlon o f i n f e r t i l i t y by w a y o £ a l t e r i n g the m a c r o m o l e c u l a r ore i z a t l o ~ of s p e r m corn n e n t s d / o r ~ e d i a in ieh sperm mo sentlaily, a phosphollpid molecule embodies a glycerol (or y other cohol) f a t t y a c i d (e.g. o l e l c a c i d d p tle a c i d ) p h o s p h o r i c acid d base such a s e h o i l n e or eth 1 he. Interference i~ y o f these ietles leads to a b n o r m a l p h o s p h o l l p i d ~e boiism.
J UNE 1978 VOL. 17 NO. 6
5¸2¸5¸i
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89 ~ 96
3751z 142 902~ 110
650~ 520
6391~ 290 361% 176 917+_ IC~3 78~ 80 930+_ i35
7545~ ~18 4 213 67~ 90 772+_ 95 1 !42
7470~ 212 40~ 120 66% 79 705! 72 1375~ 11t
699~ 211 387% ~35 6~.,+ 70 700+_ 85 t400+_ 150
3920+_ 3~ 175% IC~
521% 314 372% 28~ 2605~ 140 1!7 125 67~ 76~ 75% 70 65 3i 50 59~ 5g~ 257~ 45~ 104 39 26 40 1425~ 1 4 4 % 167-5& 15t5~ 210 66 105 57
6925+_ 581~ 180 360
* gicrograa of phospholipi•/gmof tissue weight. ~t~ndard deviation of the me~
Phospholipid Fractions: ~hospha¢i~l choline Phosphatidyl eth~ol~ne Phosphatidyl serlne Phosphati~yl inositol Sphingo~el~n
Control
~p.
i45 14!% 118 5}~_ 42 !4% 37 174~ 75
Changes in total phos~holipid and various ~hss~hospholipid fractions of testis, epidldy~ and vas deferens after ~phs-chlorohydrin ~istration
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osphatidyl choline d phosphatidyl e ol ne have been re rted to be involved in the d r e s p l r a t i o ~ O f sper tozoa b i n o s l t o l has a stimulatory On mO t i l l t y p and E l > c o l y s i s o sper~ato a (I~)Decrease of choline, phosp h a t i d y i eth el i~e d phosphatidyl o s l t o l in the C da s d vas d e f e r e n s in the p r e s e n t I n v e s t i N a t l o n le the ...d.e e r s e motility (9)d respi ra on (16)in cauda is vas A decrease phosphatldyl choline in rat epidld is a f t e r d r U g a d m i n i s t r a t i o n h a s also been by M o r r i s . (i?). observatlons conclusively sugNest at d e c r e a s e in the moie ..... tle8 i6 r e l a t e d to e c r e a s e in s~e me t i l l .
e c o n c e n t r a t l o n o f glycine, actlve p r e c u r s o r of eth el e r plasma increases a~ter chlo hydr treatment ( 1 6 ) S i n c e ...eth . .....o l a m i ~ e is a base component o f eth famine i n c r e a s e in N! Ii lead i n c r e a s e in eth el e and a d e c r e a s e phosphat y! e i i e. A decrea In pho atlayl ....i d phosNatidyl e el n e in c da epidid is d d e f e r e n s a f t e r t r e a t m e n t m a y be p a r t l y r e s p o n s i b l e for i b i t i o n of m o t i l i t y d since these have been I m p i l c a t e d in spe meta l i 6 m (i2). A d e c r e a s e in p h o s p h a t i d y l i tel theels h a s 80 been r e p o r t e d in e c d a epldld is a f t e r drug a d m l n i ~ t r a t l o n
().
A p e r t i n e n t qu e s t l o n ich arl s e s ! 8 h e w a l ph ....ch I o r o hyd !blt~ pld b l o s y n t h e a l e s p e c l f l c i y in the c d a epldid Is. i n f e r t i l i t y ~ hrs after drus admlnist %ion, m a m ace u l a t i o n of the d r u a in the cauda epldid Is a f t e r ~8 h r s d s u b s e q u e n t ch Ees in the c a u d a p h o s p h o l i ~ i d s 6u est that the d ~ N i n t e r f e r e s in phosphollpld i n the c da s an~ deferene, in p h o s p h o l i p i d g l y c e r o l en the ta ilc p a y r e ! l o w e d by its p h o s p h o r y l a t l o n . sslbl$ the ~ glycerol in the testls e p l d i d is c o m p l e x a f t e r c h l o r o h y d r treatment goes in a n o r m y b e c a u s e the g l y c e r o l l e v e l r e m a i n s al (19). it is quite to b e l i e v e the h y thesls h ~. (5) that ph c h l o r o h y d r l n Nets phosphorylated %hi8 , DL l - c h l o r o l-dee phosphate, a c t s as a i i tot Of phosphate because of I t s t al B il ty to the no al s u b s t r a t e of the enz e, 5-phosphate.
Implicitly, these ch ges y be due t o m e c h a n i s m suggested h ri e a l. (~)in pho spho I p i d 8 and ener g era ring systems the c d a e p l d l d d vas after drug
528
JUNE 1978
V O L . 17 N O . 6
CONTRACE
ION
m i n i s t r a t i o n could alter the c o m p o s i t i o n of the l u m i n a l vesl cl e or i air th elf for mation ( c da epidld al fluid cont s n u m e r o u s lu hal v e s i c l e s o to m nta e i n t e g r i t y of s p e r m p l a s m a m e m b r ( 1 2 ) ) i n f l u e n c i n g the physlco-chemlcal c h of the s p e r m p l a s m a m e m b r a n e d fertilIz pots i a i l t i e s 0£ s p e r m a t o z o a as a consequence. a l y s i s of these luminal v e s l c i e 8 from c da epldid is d Va8 clefstems d spe plasma m e m b r e after c h l o r o h y d r l n treatmen needs rther vest s.
e thors are th ful to Prof. B . R . a, Head, Depar tmen t o f oph slc s, Pan jab U n i v ty dig p fo r p d i n g l a b o r a t o r y faciiitles. Critical s u g g e s t i o n s from . L L o r i a u x (NIH) . T. n are E r a t e f u l l y ac owledged.
I.
icsso~,
R.J. 5897: A p o s t - t e s t i c u l a r tifertiilty rst nual M e e t l n g of the Boc. for the S t u d y of Reprod. d e r b i i t Univ. Nashville, Tenn, 1968.
2.
Ericsson, R.J. and er V.F. Male anti f e r t i l i t y comunds o Io sic a I pro p e r tl e s o f U - 5 8 97 U-15 646. J. Reprod. Fert. 2 1 : 2 6 7-2 73, i970.
3.
Gunn, S.A. d Gould, T. C Speci ficlty o f the v a s c u l a r system of the male r e p r o d u c t i v e tract. J. Reprod. Fert Suppl. i0: 75-95, 1969.
~.
Jones, A.R., Davies, P., Edwards, K. a n d Jackson, H. t l f e r t i l l t y effects d m e t a b o l i s m of a l p h a d in e rat. N a t u r e 224: 83, 1969.
5-
Mob H., S u t e r D.A.Io, o odin , P.D.C., its, and Ridley, D.D. I d e n t i f i c a t i o n of the ochemlcal p r o d u c e d by a l p h a - c h l o r o h y d r i n in s ~ e r m a t o z o a . Nature 75 77,
6.
S ojlik, E. and 3-ch 1 o ro-I, 2-p ro 2:
7-
8.
,
I.G.
ang, M.C. t i f e r t i l i t y a c t i v i t y of n e dl o i on mal e rat s. o i. R e pro d.
1970.
Vi ckery, B.H., ickson, G. I. and nne t t, J.P. Mech ism of t i f e r t i l i t y action of low doses of p h a - c h l o r o h driny in t h e male rat. J. Reprod. Fert. 38: i-iO, 1974. glmayr,
J.K.
Al
a-ch lorohyd rln induc e d ch an~e s in m - i n o a l t o l and PG a, and
the d i s t r i b u t i o n of f r e e
JUNE 1978 VOL. 17 NO~ 6
5
S
s o f phosphatld
the
S. 9-
p Life .
oi.
~ep
la, J . A . Sei. 8 :
d. ...ii .
ositol the r a t 600, 1974.
593
irago n a d 1969.
real e
t i f e r ill i ty
en t.
48,
I c h , j . , L e e s , M. d ~Io me tho d for e isol at n and
509,
epldid
from 1957.
tlssue,
St lex, G.H. A sl e p u r ! f l c a t i o ~ o f to t j. . . . . oi era. 2 2 6 497-
II.
M a r i ± e t t i , G.V. romato~raphlc sep cation d ysis o f phosphatldes. 3: 1-15, 1962.
12.
g l m a ~ r , d .K. Meta l i c ch ges In sper~ato a durlng epidid al tr si tion: H a n d b o o k o f (e d. Gtr e e .b , R . O O A s t w o o d ~, E . B . ) e eric ysioi. Soc e in~to~, D.O. 437-449, 1975.
13.
rner,
M.A.
fertillty
the rat. 14.
Effects
£ spermato
J.
d
of
ation, IdentlflJ. p i d ..... Re ... s.
alpha a of
Fert.
h!orohydrin u n the c da e p i d l d is o 2 4 : 2 6 7 - 2 69, 1 9 7 1 the
Crabo, B. d De i gr en, L . A . aipha- chlorohyd n mice 30: 1 6 1 - 1 6 3 , 1 9 7 2
d
s trl but io n o f i 4 Crats. j. Fert.
15
L a r d y , H A. and i i ! i p s , P.H. e interreiation of o dative d giycolytl e s s e 8 as s o u r c e o f e n e r g y for bull sp .e r m a. t o z o a.. .. . y s l o l , i"' '-33~ 60~ ...6!3, .. 1941.
16.
O o d m ...., P . D . C. d it e, .Q. S t u d ies on th e . J male t i f e r t i l i t y d r u g 3 - c .......... 1, 2- ..... R e p r o d . F e r t . 32: 337, 1973.
!7.
M o r r i s , S. , D in .... n n e t t , A., o odm , P. a n d .... ~h ..... l o r ' oh~ ....... ' .......... .....t.~,,e, I.G. E f f e c t of a i..pha--c drin the I n c o r of tO the p h o s p h o l i p l d s and g l y c e phospho r y l c h o l l n e o £ rat e p l d i d is. P r o . ......s t. o e~. C . 8: 69, 1 9 7 6 .
18.
S t e t t e n , D.W. o l o g i c a l r e l a t i o n s h i p s of c h o l i n e e t h a n o l ine r e l a t e d corn u n d s . j. ol. Chem. 1 : ! 43 .... 1 ~ 1941
19.
H o d g e n , G.D. e ~ale rat
~3 0
tal glycerol J. R e p r o d .
in rt
the
excurrent : 27'7:2
JUNE
ducts ,
1978
of .
VOL.
17 N O . 6