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Pituitary Gonadotrophins (Follicle-Stimulating Hormone and Interstitial Cell-Stimulating Hormone) and Seminal Vesicular Fructose After Long-Term Vasectomy in Bulls
Vol. 22, No.9, September 1971
FERTILITY AND STERILITY
Printed in U.S.A.
Copyright' 1971 by The Williams & Wilkins Co.
PITUITARY GONADOTROPHINS (FOLLICLE-STIMULATING HORMONE AND INTERSTITIAL CELL-STIMULATING HORMONE) AND SEMINAL VESICULAR FRUCTOSE AFTER LONG-TERM VASECTOMY IN BULLS A. M. RAKHA, PH.D.,
AND
G. IGBOELI, PH,D,
National Council Scientific Research, Physiology Laboratory, Chilanga, Zambia
;
Current interest in vasectomy as a method of sterilization requires a thorough investigation of its after effects. In a previous report 1 it was suggested that the reduced rate of sperm flow in the epididymis of chronically vasectomized bulls may by some unknown mechanism decrease the rate of spermatogenesis. The possibility of an integrating factor regulating spermatogenesis, epididymal sperm flow, storage, and resorption has also been suggested. 2 Whether this regulation is hormonal, neural, or physical is not clear, and it is intended by this study to investigate the possible role of gonadotrophins in this process. The pituitary glands and seminal vesicles were collected from five chronically vasectomized (1-5 years vasectomy), and four comparable intact bulls soon after slaughter. Each pituitary was trimmed, weighed, and the anterior lobe was frozen on Dry Ice, then freeze-dried (FD), ground to a fine powder, and stored in a desicator. The follicle-stimulating hormone (FSH) and interstitial cell-stimulating hormone (ICSH) determinations were run as single assays (2 + 1 design) on Sprague-Dawley female rats and the potency of the preparations were calculated. 3 FSH was determined by the augmentation method; 4 four animals were used per group, and each received 20 LV. of human chorionic gonadotrophin (HCG). The standard doses used were 0.1 and 0.2 mg. of NIH-FSH-P1; saline extracts of 35-50 mg. of pituitary powder Received for publication March 22, 1971.
represented the unknown. The total dose was 3 mI. (6 X 0.5 mI.). ICSH was determined by the ovarian ascorbic acid depletion method. 5 The standard doses used were 2 ,."g. and 4 ,."g. of NIH-LH-B6; a saline extract of 0.6 mg. of pituitary powder represented the unknown. Four animals were used per group. All preparations were given as a single intraperitoneal injection (0.5 mI.) and the animals killed 4 hr. later, both ovaries bulked, and the ascorbic acid concentration determined. 6, 7 The degree of depletion was estimated by reference to the mean value of a control group. The fructose concentrations of the seminal vesicles were determined. 8 The present results showed no differences in the pituitary FSH and ICSH concentrations or content between intact and vasectomized bulls. In their previous report, Igboeli and Rakha observed that chronic vasectomy resulted in a slight reduction in the rate of spermatogenesis. Although the present findings would tend to rule out hormonal intervention in this process, the possibility of an alteration in the relationship between the rates of ICSH and FSH synthesis and/ or release, or an alteration in the response of the testes following vasectomy cannot be overlooked. However, when androgen production of the testes was assessed by the concentration of fructose in the seminal vesicles, no differences were observed. This is in agreement with previous findings of Skinner and Rowson. 9 In addition, how-
581
582
Vol. 22
RAKHA AND IGBOELI
TABLE 1. Pituitary FSH and ICSH Levels and Seminal Vesicular Fructose in Vasectomized Animals FSH activity in terms Group
of NIH-FSH-S,
SV* fructose
Concentration
(FD)
mg.!lOO gm.
J-llI.!m/i.
222 233 233 256
1.66(0.53-3.15) 1.60(0.36-3.35) 1.63(0.96-3.02) 1.20(0.24-2.16)
Mean ± S.E. Vasectomized
236 ± 7.2 330 259 181 256
1.52 ± 0.11 1.89(0.61-2.89) 0.87(0.06-0.88) 1.06(0.08-1.39) 1.94(0.80-3.98) 1.66(0.67-3.65)
Mean ± S.E.
256.5 ± 30.4
1.48 ± 0.22
Intact
Content
of
NIH-LH-S,
A
Concentration (FD) J-llI.!mg.
J-llI.
0.10 0.12 0.17 0.17
28.52(2.85-61.60) 17.92(10.70-27.18) 10.94(3.50-13.3) 8.88(0.96-9.79)
14488.2 6505.0 1717.6 1696.1
J-llI.
843.3 580.8 255.9 229.21
)-
ICSH activity in terms
477.3 ± 145.80 1032 0.25 724 0.15 524 0.16 565 0.11 354 0.10 640 ± 114.40
Content
Ratio
ICSHI A
FSH
0.23 0.29 0.24 0.22
17.18 11.20 6.71 7.40
16.57 ± 4.43 7.85(3.75-11.68) 6.38(1.6-7.29) 9.95(2.71-10.45) 21.27(7.36-35.35) 30.39(7.01-69.90)
6101.7 ± 301.60 0.30 4286 0.19 5308 0.22 4915 6190 0.28 0.22 5045
10.90
15.17 ± 4.62
5149 ± 309.75
4.15 7.33 9.39 10.96 18.31 10.25
* SV refers to the concentration of fructose/100 gm. of the seminal vesicles.
ever, these authors reported a significant reduction in the fructose and citric acid content of the ampullae of vasectomized pubescent animals, which was attributed to a purely physical interruption of the normal passage of testosterone along the vas deferens. Further work is required to investigate the physical and neural factors which may result from vasectomy and which may operate to alter the normal equilibrium between spermatogenesis, sperm flow, and sperm resorption. SUMMARY
The concentrations and contents of pituitary gonadotrophins (FSH and ICSH) and seminal vesicular fructose, in five chronically vasectomized (1-5 years vasectomy) and four intact bulls, were determined. The present results showed that vasectomy in bulls did not alter the pituitary FSH or ICSH concentration and content. Similarly, there was no effect on androgen production of the testes as expressed in terms of fructose concentration in the seminal vesicles.
Acknowledgments. The authors are grateful to D. K. Dutta-Roy for his advice in the statistical analysis; Messrs. D. Lungu, J. Parshotam, and S. Chilinda for technical assistance; and to the Endocrine Study Section of the National Institutes of Health for the standard pituitary hormone preparations. REFERENCES 1. IGBOELI, G., AND RAKHA, A. M. Bull testicular and epididymal functions after long-term vasectomy. J Anim Sci 31 :72, 1970. 2. AMANN, R. P., AND ALMQUIST, J. O. Reproductive capacity of dairy bulls: VI. Effect of unilateral vasectomy and ejaculation on sperm reserves; aspects of epididymal physiology. J Reprod Fertil 3:260, 1962. 3. GADDUM, J. H. Simplified mathematics for bioassays. J Pharm Pharmacal 5:345, 1953. 4. STEELMAN, S. 1., AND POHLEY, F. M. Assay of follicle stimulating hormone based on the augmentation with HCG. Endocrinology 53:604, 1953. 5. PARLOW, A. F. "Bioassay of pituitary luteinizing hormone by depletion of ovarian ascorbic acid." In Human Pituitary Gonadotrophins, Albert, A. Ed. Thomas, Springfield, TIl. 1961, pp. 300-310. 6. MAIKEL, R. P. A rapid procedure for the determination of adrenal ascorbic acid. Application of the Sullivan and Clarks method to tissue analysis. Analyt Biochem 1:498, 1960. 7. ROBERTSON, H. A., AND RAKHA, A. M. The se-
.
,
.
• ..
September 1971
LONG-TERM VASECTOMY IN BULLS
quence, time and duration, of the release of follicle stimulating hormone and luteinizing hormone in relation to oestrus and ovulation in the sheep. J Endocr 35:177, 1966. 8. LINDNER, H. R., AND MANN, T. Relationship between the content of androgenic steroids in the
583
testes and the secretory activity of the seminal vesicles in the bull. J Endocr 21:241, 1960. 9. SKINNER, J. D., AND RoWSON, L. E. A. Some effects of unilateral cryptorchism and vasectomy on sexual development of the pubescent ram and bull. J. Endocr. 42:311, 1968.
FERTILITY AND STERILITY
Printed in U.S.A.
Copyright' 1971 by The Williams & Wilkins Co.
PITUITARY GONADOTROPHINS (FOLLICLE-STIMULATING HORMONE AND INTERSTITIAL CELL-STIMULATING HORMONE) AND SEMINAL VESICULAR FRUCTOSE AFTER LONG-TERM VASECTOMY IN BULLS A. M. RAKHA, PH.D.,
AND
G. IGBOELI, PH,D,
National Council Scientific Research, Physiology Laboratory, Chilanga, Zambia
;
Current interest in vasectomy as a method of sterilization requires a thorough investigation of its after effects. In a previous report 1 it was suggested that the reduced rate of sperm flow in the epididymis of chronically vasectomized bulls may by some unknown mechanism decrease the rate of spermatogenesis. The possibility of an integrating factor regulating spermatogenesis, epididymal sperm flow, storage, and resorption has also been suggested. 2 Whether this regulation is hormonal, neural, or physical is not clear, and it is intended by this study to investigate the possible role of gonadotrophins in this process. The pituitary glands and seminal vesicles were collected from five chronically vasectomized (1-5 years vasectomy), and four comparable intact bulls soon after slaughter. Each pituitary was trimmed, weighed, and the anterior lobe was frozen on Dry Ice, then freeze-dried (FD), ground to a fine powder, and stored in a desicator. The follicle-stimulating hormone (FSH) and interstitial cell-stimulating hormone (ICSH) determinations were run as single assays (2 + 1 design) on Sprague-Dawley female rats and the potency of the preparations were calculated. 3 FSH was determined by the augmentation method; 4 four animals were used per group, and each received 20 LV. of human chorionic gonadotrophin (HCG). The standard doses used were 0.1 and 0.2 mg. of NIH-FSH-P1; saline extracts of 35-50 mg. of pituitary powder Received for publication March 22, 1971.
represented the unknown. The total dose was 3 mI. (6 X 0.5 mI.). ICSH was determined by the ovarian ascorbic acid depletion method. 5 The standard doses used were 2 ,."g. and 4 ,."g. of NIH-LH-B6; a saline extract of 0.6 mg. of pituitary powder represented the unknown. Four animals were used per group. All preparations were given as a single intraperitoneal injection (0.5 mI.) and the animals killed 4 hr. later, both ovaries bulked, and the ascorbic acid concentration determined. 6, 7 The degree of depletion was estimated by reference to the mean value of a control group. The fructose concentrations of the seminal vesicles were determined. 8 The present results showed no differences in the pituitary FSH and ICSH concentrations or content between intact and vasectomized bulls. In their previous report, Igboeli and Rakha observed that chronic vasectomy resulted in a slight reduction in the rate of spermatogenesis. Although the present findings would tend to rule out hormonal intervention in this process, the possibility of an alteration in the relationship between the rates of ICSH and FSH synthesis and/ or release, or an alteration in the response of the testes following vasectomy cannot be overlooked. However, when androgen production of the testes was assessed by the concentration of fructose in the seminal vesicles, no differences were observed. This is in agreement with previous findings of Skinner and Rowson. 9 In addition, how-
581
582
Vol. 22
RAKHA AND IGBOELI
TABLE 1. Pituitary FSH and ICSH Levels and Seminal Vesicular Fructose in Vasectomized Animals FSH activity in terms Group
of NIH-FSH-S,
SV* fructose
Concentration
(FD)
mg.!lOO gm.
J-llI.!m/i.
222 233 233 256
1.66(0.53-3.15) 1.60(0.36-3.35) 1.63(0.96-3.02) 1.20(0.24-2.16)
Mean ± S.E. Vasectomized
236 ± 7.2 330 259 181 256
1.52 ± 0.11 1.89(0.61-2.89) 0.87(0.06-0.88) 1.06(0.08-1.39) 1.94(0.80-3.98) 1.66(0.67-3.65)
Mean ± S.E.
256.5 ± 30.4
1.48 ± 0.22
Intact
Content
of
NIH-LH-S,
A
Concentration (FD) J-llI.!mg.
J-llI.
0.10 0.12 0.17 0.17
28.52(2.85-61.60) 17.92(10.70-27.18) 10.94(3.50-13.3) 8.88(0.96-9.79)
14488.2 6505.0 1717.6 1696.1
J-llI.
843.3 580.8 255.9 229.21
)-
ICSH activity in terms
477.3 ± 145.80 1032 0.25 724 0.15 524 0.16 565 0.11 354 0.10 640 ± 114.40
Content
Ratio
ICSHI A
FSH
0.23 0.29 0.24 0.22
17.18 11.20 6.71 7.40
16.57 ± 4.43 7.85(3.75-11.68) 6.38(1.6-7.29) 9.95(2.71-10.45) 21.27(7.36-35.35) 30.39(7.01-69.90)
6101.7 ± 301.60 0.30 4286 0.19 5308 0.22 4915 6190 0.28 0.22 5045
10.90
15.17 ± 4.62
5149 ± 309.75
4.15 7.33 9.39 10.96 18.31 10.25
* SV refers to the concentration of fructose/100 gm. of the seminal vesicles.
ever, these authors reported a significant reduction in the fructose and citric acid content of the ampullae of vasectomized pubescent animals, which was attributed to a purely physical interruption of the normal passage of testosterone along the vas deferens. Further work is required to investigate the physical and neural factors which may result from vasectomy and which may operate to alter the normal equilibrium between spermatogenesis, sperm flow, and sperm resorption. SUMMARY
The concentrations and contents of pituitary gonadotrophins (FSH and ICSH) and seminal vesicular fructose, in five chronically vasectomized (1-5 years vasectomy) and four intact bulls, were determined. The present results showed that vasectomy in bulls did not alter the pituitary FSH or ICSH concentration and content. Similarly, there was no effect on androgen production of the testes as expressed in terms of fructose concentration in the seminal vesicles.
Acknowledgments. The authors are grateful to D. K. Dutta-Roy for his advice in the statistical analysis; Messrs. D. Lungu, J. Parshotam, and S. Chilinda for technical assistance; and to the Endocrine Study Section of the National Institutes of Health for the standard pituitary hormone preparations. REFERENCES 1. IGBOELI, G., AND RAKHA, A. M. Bull testicular and epididymal functions after long-term vasectomy. J Anim Sci 31 :72, 1970. 2. AMANN, R. P., AND ALMQUIST, J. O. Reproductive capacity of dairy bulls: VI. Effect of unilateral vasectomy and ejaculation on sperm reserves; aspects of epididymal physiology. J Reprod Fertil 3:260, 1962. 3. GADDUM, J. H. Simplified mathematics for bioassays. J Pharm Pharmacal 5:345, 1953. 4. STEELMAN, S. 1., AND POHLEY, F. M. Assay of follicle stimulating hormone based on the augmentation with HCG. Endocrinology 53:604, 1953. 5. PARLOW, A. F. "Bioassay of pituitary luteinizing hormone by depletion of ovarian ascorbic acid." In Human Pituitary Gonadotrophins, Albert, A. Ed. Thomas, Springfield, TIl. 1961, pp. 300-310. 6. MAIKEL, R. P. A rapid procedure for the determination of adrenal ascorbic acid. Application of the Sullivan and Clarks method to tissue analysis. Analyt Biochem 1:498, 1960. 7. ROBERTSON, H. A., AND RAKHA, A. M. The se-
.
,
.
• ..
September 1971
LONG-TERM VASECTOMY IN BULLS
quence, time and duration, of the release of follicle stimulating hormone and luteinizing hormone in relation to oestrus and ovulation in the sheep. J Endocr 35:177, 1966. 8. LINDNER, H. R., AND MANN, T. Relationship between the content of androgenic steroids in the
583
testes and the secretory activity of the seminal vesicles in the bull. J Endocr 21:241, 1960. 9. SKINNER, J. D., AND RoWSON, L. E. A. Some effects of unilateral cryptorchism and vasectomy on sexual development of the pubescent ram and bull. J. Endocr. 42:311, 1968.