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Cyproterone acetate and seminal vesicles in the regulation of male fertility
CONTRACEPTION
CYPBOTEBOHE ACETATE AND SEMINAL VESICLES IN TBE REGULATION OF BALE FEETILITY
S.
K, Nayyar and Tultul Chosh*
University Colleges of Science & Technology Department of Physiology, University of Calcutta, Calcutta-700 009, India
ABSTRACT
The effect of continuous release of microquantities of cyproterone acetate from subcutaneously implanted silastic capsules on the fertility of surgically manipulated seminal vesicles of rat has been studied. The experimental animals showed infertility for 3 weeks, as a consequence of removal of seminal plasma, following which normal fentility returns as efficiently as that of operated,control. However, a significant decrease in weight of the testes and epididymidee, were recorded, It appeans that seminal vesicles can synthesize seminal plasma avnn at very low levels of androgen in-ths presence of cyproterone 'acetate.
Accepted for publication January 11, 1980 * Reprint
FEBRUARY
requests
to:
1980VOL.21N0.2
Tultul
Ghosh
183
CONTRACEPTION
INTRODUCTION
Early experiments of Steinach ( 1894) and Walker ( l.900) showed that mating and also facundity are possible in rats after excision of either the prostate or the seminal vesicles (for reference see 1). Later Nalbandov (2) also reported that both the boars and rats did retain their fertility in However the the absence of seminal vesicles and rostate. st$.mlausefulness of seminal plasma In exer e ing a distinct ting effect on sperm motility has been repeatedly suggested In normal fertile male rats, surgical removal by Mann (1). of plasma from seminal vesicles make the animals infertfle and after a time lag of 3 weeks normal fertility returns (3). This emphasises that a certain amouut of vesicular plasma Is needed for retaining male fertility and the ssmlnal vesicles possibly take about 3 weeks to accumulate the amount of required plasma. Cyproterone acetate (CPA), an antiandrogen (41, was found to decrease the uptake of radioactive testosterone in male accessory glands (5 6) and it can also prevent the accwnulation of both testosterone and dehydrotestosterone in the seminal vesicles (7). Steinbeck & &,. (8) have reported a suppression of accessory sex glands and fertility in rats by both high and low dose of CPA. Furthermore, failure of CPA in producing antifertility effects in rats has also been observed by several workers (9,101. Our experimental whether t i)
ii)
184
design
was aimed at investigating
CPA can alter the expected time for restoration of seminal plasma thereby affecting the fertilfty of seminal plasma-removed rats, and CPA can suppress
accessory
sex glands.
FEBRUARY 1980 VOL. 21 NO. 2
CONTRACEPTION
MATERIALS ANDMSTHODS
Sexually mature colony bred albino male rats (200-350 g) were kept in good husbaudry conditions of 12 h light per day and fed on a standard rat diet. CPA was kindly supplied by Schering AG Berlin. Four silastic capsules containing CPA (111, were implan(30 mg eachj, a s prepared by Prasad & a. ted in each rat subcutaneously under light ether anesthesia. The animals were sorted Into different groups and were left In lace for 90, 120 and lS0 days, respectively (Table I, II), The vesicular ! efore they were operated on for plasma removal. plaza was removed by squeezing out the vesicular content through a small cut made on the body of the seminal vesicles. Fertility tests were made on the animals on a weekly basis until Cyclic feanales on the day of their they were sacrificed. vaginal proestrus were paired overnight with the test animals. Sperm positive vaginal smear was designated as day 1 postcoitun Unilateral uterine traumatlzation was done on the mated (p.c). females on day 5 p.c. for inducing decidual cell reaction (DCR). Laparotomy was performed on day 10 p.c. to confirm pregnancy or pseudopregnancy by examining the presence of implantation site (s) or DCR, respectively.
Forty days following seminal plasma removal, the animals were sacrificed and the testes, epididymides, seminal vesicles, prostates and coagulating glands were removed weighed and compared with the controls using student’s E test. To ascertain the specific effects of CPA non-operated ZPA 1.n addition to operated capsules-bearing animals were included, animals with empty capsules only for control purposes.
HEWJLTSANDDISCUSSION It was consistently observed that continuous release of microquantities of CPA from implanted silastlc capsules did not affect the fertility of the males at all. Mating records of both operated control and experimental animals showed that during the first 3 weeks following surgical manipulation of seminal plasma the mated females became pseudopregnant as evident from t he usual degree of DCR (1345.8 2 91.3 vs 107.5 2 12.2) without showing any implantation site (s) in the non-traumatized uterine horn. Thus indicating intact sexual
FEBRUARY 1980 VOL. 21 NO. 2
185
160
150
120
I.20
90
90
190
190
160
160
130
_
130
1~~
412.e56.1 (P< 0.01)
142.e3.1 (PC 0.001)
X39.%6.1
42O.s+l9.8 144.e7.4 (PL 0.001) (P< 0.01) 679.e21.8
163.e10.2
148&8.8
--
40.a5.1
138.e15.7
lJi3.ti8.3
45.a3.1
154.0+_11.8122.e15.2
140.8+,10*7121&4.0
44.725.9
47.722.7
X%4.2&9.9
120.*13.1
48.4to.9
44.e8.2
51.e6.7
123.e7.0 l25.*9.3
46.821.8
48.a2.4
l22&17.3
122.e7.8
Prostate Coagulating wt;(mg) gland wt (mg)
1.53.el2.7 l24.ti8.7
145.ti2.7
'ol~j6 139.~5.9 .
193.829.8
149.7212.0 (P < 0.05)
677.2 $30.0 184.725.6
~~~~~j2 .
690.%21.5
500.e28.4 (PKO.01)
:%: at (mg)
* Animals tith 4 CPA capsules. ++ Animals with 4 empty capsules. *** Days followingcapsules implantation.
Operated+ 4CPA capsules (8)
Operated control** (8)
Non-operated control* (8)
Operated+ 4CPA capsules (8)
Operated control** (8)
Non-operated control* (8)
Operated+ 4CPA capsules (8)
Operated control** (8)
Non-operated control* (8)
Epldidymal wt (mg)
t Organ wel&t (mg/lOO gm bw) of seminal plasma-removedrats following exposure to CPA (Mean + SE)
Period of Testicular Treatment Period of (I?o.of animals) rest before ;o"p";~&e wt (mg) operation (Days+**) (Days)
TABLE I
3 8 2
ff
3
8
1W0(100%)
8/8(100%)
90
120
120
150
150
Operated + 4CPA ca sules P8)
Operated control (8)
Operated + 4CPA ca sules P8)
Operated control (8)
Operated + 4CPA ca sules P8)
8/8(100%)
8/8(100%)
8/8(100%)
5th (P) --I__
8/8(100%)
8r(8(100%)
1W10(100%)
8/8(100%)
8/8(100%)
10/10(100%) 10/10(100%)
8/8(100%)
10/10(100%) 10/10(100%) 10/10(100%)
e/8(75%)
8/8(100%)
8/8(100%)
8/8(100%)
8/8(100%)
10/10(100%) 10/10(100%) lO/lO(l()()%) 8/8(100%)
8/8(x30%)
: Pregnant P Ps : Pseudopregnant Non-operated controlled animals of all the three groups showed fertile matings throughout the experimental period .
9/10(90%)
8/8(100%)
lWlO(lOO%)
8/8(100%)
90
Operated control (8)
II :
Fertility Scoring ____-__C___---_-__-----r. Mating records (week following operation) Treatment Period of 3-k (No. of animals) rest before 1s wee 0 eratlon (Ps) (Ps) (Ps) (P) PDays) _-
TABLE
F Q Lp
8.
CONTRACEPTION
activity capacity
flthout retaining the potential fertilizing Pregnancy was, however, recorded of the spermatozoa. from the 4th week onwards (Tab1e11). It had been earlier suggested (3) that a certain amount of seminal plasma is needed for retaining male fertility, and in the plasmaremoved animals, exogenously administered gonadotrophlns or testosterone could restore fertility by stimulating the If this hypothesis secretory activity of seminal vesicles. is true, then the expected time for the restoration of seminal plasma in CPA capsules-bearing animals should alter, as CPA is known to act by competitive inhibit ion on endogenously and exogenously administered testosterone at their target But in our experimentation, the mating records organs (12). revealed that in both operated control and experimental group, fertility was restored after 3 weeks following surgical manipulation of seminal vesicles. Thus CPA had no impact whatsoever in the time factor involved in the restoration of seminal plasma following its removal. However, long-term loss of testicular and exposure to CPA resulted in significant epididymal weight and the other accessory sex organs remained In the rat CPA has been shown to unaffected (Table I). modify the release of gonadotroph&s (13)) secretory activity of accessory sex glands( 11)) and mating sequences ( 14). However since the CPA, in our experimental model fails to reduce ihe fertilizing capacity of the rats havink surgically manipulated seminal vesicles, it is, therefore, tempting to suggest that seminal vesicles can synthesize seminal plasma even ,at very low levels of androgen in the presence of CPA.
ACKNOWLEDGEMENT The authors sincerely wish to acknowledge the help, co-operation and inspiration extended to them by Dr.B.N.Kolay, Dr.(Mrs.) J.Kolay, Dr.A.K.Pal and Dr.(Mrs.) M.Gupta. Thanks are also due to Director General, Council of Scientific and Industrial Research, New Delhi, for financing this project.
188
FEBRUARY 1980 VOL. 21 NO. 2
CONTRACEPTION
REPeRENCES 1.
Mann, T. : The Biochemistry of Semen and of the Male Reproductive Tract. John Wiley & Sons, Inc., New York, 1964
2.
Nalbandov, A.V. : Reproductive Physiology. and Company, San Francisco, 2958
3.
Chatterjee, A. and Pal, A.K. : Seminal vesicles and its Importance In the regulation of male fertility. Andrologia 10 (4) : 321, 1978
4.
R, I,, Elger, W., Nemann, F., Von Berswordt-Wallrabe, Steinbeck H., Hahn, J.D. and Kramer, M. : Aspects of androgen-dependent events as studied by antlandrogens. Recent Progr. Horm. Res. X : 337, I970
5.
Garabieta G $%i Ji' z$ !%%th”‘6 : Effeci o;‘c$P”&t&e acetate'on'Z%testoster&e'uptake and enzyme synthesis by ventral prostate of the rat. Endocrinology 84 I 1330, I.969
6.
Stern, J.M. and Eisenfeld, A.J. : Distribution and metabolism of 3H-testosterone in castrated male rats: Effects of cyproterone, progesterone and unlabelled testosterone. Endocrinology 88 t 1117, 1971
7.
Stern J.M. and Eisenfeld, A.J. : Androgen accumulation and binding to macromolecules in seminal vesicles : inhibition by cyproterone. Science 166 t 233, 2369
8.
Steinbeck, H., Mehring, M. and Neumann, F. : Comparison of the effects of cyproterone cyproterone acetate and oestradiol on testicular fun&ion, accessory sexual glands and fertility in a long term study of rats. J. Reprod. Fert. 26 : 65, 1971
9.
Chatterjee, A., Ray, P., Gupta, M., Pal, A.K. and Kolay, A.R. t Cyproterone acetate : I. Microquantity releasing device of cyproterone acetate and Its failure in inducing functional sterility in male rats. Andrologia 9 (1) t 70, 1977
FEBRUARY
1980VOL.21N0.2
W.H.FTeeman
189
CONTRACEPTION
10.
Back, D.J. and Shenton, J.C. I Release of ticroquantlties of cyproterone acetate from subcutaneous sllastlc capsules failure to produce an antifertility effect. Contraception 14 : 17, 3976.
11.
M. and Reddy, P.R.K. t Prasad, M.&N., Rajalakshnl Action of cyproterone aceta c e on male reproductive Hormone Antagonist. Gynec. Invest. 2 : functions. x)2, l97l/72
12.
Neumann, F. and Von Berswordt-Wallrabe R.I. L Effects of androgen antagonist cyproterone ace c ate on the testicular structure, spermatogenesis and accessory sexual glands of testosterone-treated adult hypopbysectomieed rats. J. Endocrinol. 35 I 363, 1966.
13.
Schoonees, R., Schalch D.S. and Murphy, G.P. : The hormonal effects of anglandrogen (SH-714) treatment in man. Invest. Ural. 8 : 635, 1971.
14.
effects of cyproterone acetate Agmo, A. t Differential on sexual behavior and accessory sexual glands In rabbits. Acta. Physiol. Stand. 91 I 29, 1974.
190
FEBRUARY 1980 VOL. 21 NO. 2
:
CYPBOTEBOHE ACETATE AND SEMINAL VESICLES IN TBE REGULATION OF BALE FEETILITY
S.
K, Nayyar and Tultul Chosh*
University Colleges of Science & Technology Department of Physiology, University of Calcutta, Calcutta-700 009, India
ABSTRACT
The effect of continuous release of microquantities of cyproterone acetate from subcutaneously implanted silastic capsules on the fertility of surgically manipulated seminal vesicles of rat has been studied. The experimental animals showed infertility for 3 weeks, as a consequence of removal of seminal plasma, following which normal fentility returns as efficiently as that of operated,control. However, a significant decrease in weight of the testes and epididymidee, were recorded, It appeans that seminal vesicles can synthesize seminal plasma avnn at very low levels of androgen in-ths presence of cyproterone 'acetate.
Accepted for publication January 11, 1980 * Reprint
FEBRUARY
requests
to:
1980VOL.21N0.2
Tultul
Ghosh
183
CONTRACEPTION
INTRODUCTION
Early experiments of Steinach ( 1894) and Walker ( l.900) showed that mating and also facundity are possible in rats after excision of either the prostate or the seminal vesicles (for reference see 1). Later Nalbandov (2) also reported that both the boars and rats did retain their fertility in However the the absence of seminal vesicles and rostate. st$.mlausefulness of seminal plasma In exer e ing a distinct ting effect on sperm motility has been repeatedly suggested In normal fertile male rats, surgical removal by Mann (1). of plasma from seminal vesicles make the animals infertfle and after a time lag of 3 weeks normal fertility returns (3). This emphasises that a certain amouut of vesicular plasma Is needed for retaining male fertility and the ssmlnal vesicles possibly take about 3 weeks to accumulate the amount of required plasma. Cyproterone acetate (CPA), an antiandrogen (41, was found to decrease the uptake of radioactive testosterone in male accessory glands (5 6) and it can also prevent the accwnulation of both testosterone and dehydrotestosterone in the seminal vesicles (7). Steinbeck & &,. (8) have reported a suppression of accessory sex glands and fertility in rats by both high and low dose of CPA. Furthermore, failure of CPA in producing antifertility effects in rats has also been observed by several workers (9,101. Our experimental whether t i)
ii)
184
design
was aimed at investigating
CPA can alter the expected time for restoration of seminal plasma thereby affecting the fertilfty of seminal plasma-removed rats, and CPA can suppress
accessory
sex glands.
FEBRUARY 1980 VOL. 21 NO. 2
CONTRACEPTION
MATERIALS ANDMSTHODS
Sexually mature colony bred albino male rats (200-350 g) were kept in good husbaudry conditions of 12 h light per day and fed on a standard rat diet. CPA was kindly supplied by Schering AG Berlin. Four silastic capsules containing CPA (111, were implan(30 mg eachj, a s prepared by Prasad & a. ted in each rat subcutaneously under light ether anesthesia. The animals were sorted Into different groups and were left In lace for 90, 120 and lS0 days, respectively (Table I, II), The vesicular ! efore they were operated on for plasma removal. plaza was removed by squeezing out the vesicular content through a small cut made on the body of the seminal vesicles. Fertility tests were made on the animals on a weekly basis until Cyclic feanales on the day of their they were sacrificed. vaginal proestrus were paired overnight with the test animals. Sperm positive vaginal smear was designated as day 1 postcoitun Unilateral uterine traumatlzation was done on the mated (p.c). females on day 5 p.c. for inducing decidual cell reaction (DCR). Laparotomy was performed on day 10 p.c. to confirm pregnancy or pseudopregnancy by examining the presence of implantation site (s) or DCR, respectively.
Forty days following seminal plasma removal, the animals were sacrificed and the testes, epididymides, seminal vesicles, prostates and coagulating glands were removed weighed and compared with the controls using student’s E test. To ascertain the specific effects of CPA non-operated ZPA 1.n addition to operated capsules-bearing animals were included, animals with empty capsules only for control purposes.
HEWJLTSANDDISCUSSION It was consistently observed that continuous release of microquantities of CPA from implanted silastlc capsules did not affect the fertility of the males at all. Mating records of both operated control and experimental animals showed that during the first 3 weeks following surgical manipulation of seminal plasma the mated females became pseudopregnant as evident from t he usual degree of DCR (1345.8 2 91.3 vs 107.5 2 12.2) without showing any implantation site (s) in the non-traumatized uterine horn. Thus indicating intact sexual
FEBRUARY 1980 VOL. 21 NO. 2
185
160
150
120
I.20
90
90
190
190
160
160
130
_
130
1~~
412.e56.1 (P< 0.01)
142.e3.1 (PC 0.001)
X39.%6.1
42O.s+l9.8 144.e7.4 (PL 0.001) (P< 0.01) 679.e21.8
163.e10.2
148&8.8
--
40.a5.1
138.e15.7
lJi3.ti8.3
45.a3.1
154.0+_11.8122.e15.2
140.8+,10*7121&4.0
44.725.9
47.722.7
X%4.2&9.9
120.*13.1
48.4to.9
44.e8.2
51.e6.7
123.e7.0 l25.*9.3
46.821.8
48.a2.4
l22&17.3
122.e7.8
Prostate Coagulating wt;(mg) gland wt (mg)
1.53.el2.7 l24.ti8.7
145.ti2.7
'ol~j6 139.~5.9 .
193.829.8
149.7212.0 (P < 0.05)
677.2 $30.0 184.725.6
~~~~~j2 .
690.%21.5
500.e28.4 (PKO.01)
:%: at (mg)
* Animals tith 4 CPA capsules. ++ Animals with 4 empty capsules. *** Days followingcapsules implantation.
Operated+ 4CPA capsules (8)
Operated control** (8)
Non-operated control* (8)
Operated+ 4CPA capsules (8)
Operated control** (8)
Non-operated control* (8)
Operated+ 4CPA capsules (8)
Operated control** (8)
Non-operated control* (8)
Epldidymal wt (mg)
t Organ wel&t (mg/lOO gm bw) of seminal plasma-removedrats following exposure to CPA (Mean + SE)
Period of Testicular Treatment Period of (I?o.of animals) rest before ;o"p";~&e wt (mg) operation (Days+**) (Days)
TABLE I
3 8 2
ff
3
8
1W0(100%)
8/8(100%)
90
120
120
150
150
Operated + 4CPA ca sules P8)
Operated control (8)
Operated + 4CPA ca sules P8)
Operated control (8)
Operated + 4CPA ca sules P8)
8/8(100%)
8/8(100%)
8/8(100%)
5th (P) --I__
8/8(100%)
8r(8(100%)
1W10(100%)
8/8(100%)
8/8(100%)
10/10(100%) 10/10(100%)
8/8(100%)
10/10(100%) 10/10(100%) 10/10(100%)
e/8(75%)
8/8(100%)
8/8(100%)
8/8(100%)
8/8(100%)
10/10(100%) 10/10(100%) lO/lO(l()()%) 8/8(100%)
8/8(x30%)
: Pregnant P Ps : Pseudopregnant Non-operated controlled animals of all the three groups showed fertile matings throughout the experimental period .
9/10(90%)
8/8(100%)
lWlO(lOO%)
8/8(100%)
90
Operated control (8)
II :
Fertility Scoring ____-__C___---_-__-----r. Mating records (week following operation) Treatment Period of 3-k (No. of animals) rest before 1s wee 0 eratlon (Ps) (Ps) (Ps) (P) PDays) _-
TABLE
F Q Lp
8.
CONTRACEPTION
activity capacity
flthout retaining the potential fertilizing Pregnancy was, however, recorded of the spermatozoa. from the 4th week onwards (Tab1e11). It had been earlier suggested (3) that a certain amount of seminal plasma is needed for retaining male fertility, and in the plasmaremoved animals, exogenously administered gonadotrophlns or testosterone could restore fertility by stimulating the If this hypothesis secretory activity of seminal vesicles. is true, then the expected time for the restoration of seminal plasma in CPA capsules-bearing animals should alter, as CPA is known to act by competitive inhibit ion on endogenously and exogenously administered testosterone at their target But in our experimentation, the mating records organs (12). revealed that in both operated control and experimental group, fertility was restored after 3 weeks following surgical manipulation of seminal vesicles. Thus CPA had no impact whatsoever in the time factor involved in the restoration of seminal plasma following its removal. However, long-term loss of testicular and exposure to CPA resulted in significant epididymal weight and the other accessory sex organs remained In the rat CPA has been shown to unaffected (Table I). modify the release of gonadotroph&s (13)) secretory activity of accessory sex glands( 11)) and mating sequences ( 14). However since the CPA, in our experimental model fails to reduce ihe fertilizing capacity of the rats havink surgically manipulated seminal vesicles, it is, therefore, tempting to suggest that seminal vesicles can synthesize seminal plasma even ,at very low levels of androgen in the presence of CPA.
ACKNOWLEDGEMENT The authors sincerely wish to acknowledge the help, co-operation and inspiration extended to them by Dr.B.N.Kolay, Dr.(Mrs.) J.Kolay, Dr.A.K.Pal and Dr.(Mrs.) M.Gupta. Thanks are also due to Director General, Council of Scientific and Industrial Research, New Delhi, for financing this project.
188
FEBRUARY 1980 VOL. 21 NO. 2
CONTRACEPTION
REPeRENCES 1.
Mann, T. : The Biochemistry of Semen and of the Male Reproductive Tract. John Wiley & Sons, Inc., New York, 1964
2.
Nalbandov, A.V. : Reproductive Physiology. and Company, San Francisco, 2958
3.
Chatterjee, A. and Pal, A.K. : Seminal vesicles and its Importance In the regulation of male fertility. Andrologia 10 (4) : 321, 1978
4.
R, I,, Elger, W., Nemann, F., Von Berswordt-Wallrabe, Steinbeck H., Hahn, J.D. and Kramer, M. : Aspects of androgen-dependent events as studied by antlandrogens. Recent Progr. Horm. Res. X : 337, I970
5.
Garabieta G $%i Ji' z$ !%%th”‘6 : Effeci o;‘c$P”&t&e acetate'on'Z%testoster&e'uptake and enzyme synthesis by ventral prostate of the rat. Endocrinology 84 I 1330, I.969
6.
Stern, J.M. and Eisenfeld, A.J. : Distribution and metabolism of 3H-testosterone in castrated male rats: Effects of cyproterone, progesterone and unlabelled testosterone. Endocrinology 88 t 1117, 1971
7.
Stern J.M. and Eisenfeld, A.J. : Androgen accumulation and binding to macromolecules in seminal vesicles : inhibition by cyproterone. Science 166 t 233, 2369
8.
Steinbeck, H., Mehring, M. and Neumann, F. : Comparison of the effects of cyproterone cyproterone acetate and oestradiol on testicular fun&ion, accessory sexual glands and fertility in a long term study of rats. J. Reprod. Fert. 26 : 65, 1971
9.
Chatterjee, A., Ray, P., Gupta, M., Pal, A.K. and Kolay, A.R. t Cyproterone acetate : I. Microquantity releasing device of cyproterone acetate and Its failure in inducing functional sterility in male rats. Andrologia 9 (1) t 70, 1977
FEBRUARY
1980VOL.21N0.2
W.H.FTeeman
189
CONTRACEPTION
10.
Back, D.J. and Shenton, J.C. I Release of ticroquantlties of cyproterone acetate from subcutaneous sllastlc capsules failure to produce an antifertility effect. Contraception 14 : 17, 3976.
11.
M. and Reddy, P.R.K. t Prasad, M.&N., Rajalakshnl Action of cyproterone aceta c e on male reproductive Hormone Antagonist. Gynec. Invest. 2 : functions. x)2, l97l/72
12.
Neumann, F. and Von Berswordt-Wallrabe R.I. L Effects of androgen antagonist cyproterone ace c ate on the testicular structure, spermatogenesis and accessory sexual glands of testosterone-treated adult hypopbysectomieed rats. J. Endocrinol. 35 I 363, 1966.
13.
Schoonees, R., Schalch D.S. and Murphy, G.P. : The hormonal effects of anglandrogen (SH-714) treatment in man. Invest. Ural. 8 : 635, 1971.
14.
effects of cyproterone acetate Agmo, A. t Differential on sexual behavior and accessory sexual glands In rabbits. Acta. Physiol. Stand. 91 I 29, 1974.
190
FEBRUARY 1980 VOL. 21 NO. 2
: