Abstract
This study concerned the minimum and optimum effective doses of calcium chloride needed for induction of chemosterilization in male albino rats, 30 days after a single intratesticular injection of calcium chloride (CaCl2.2H2O) solution at 2.5, 5, 10 or 20 mg per 100 g body weight per testis. There was a significant diminution in the relative wet weight of the sex organs (p<0.01), epididymal sperm count (p<0.001), plasma concentration of testosterone (p<0.01), testicular activities of D,3β-hydroxy steroid dehydrogenase (D,3β-HSD), 17β-hydroxy steroid dehydrogenase (17β-HSD) (p<0.01), glutathione S-transferase (GST) (p<0.01), superoxide dismutase (SOD) (p<0.01), and peroxidase (p<0.01), significant elevations in testicular content of malondialdehyde (MDA) and conjugated dienes (p<0.01), along with derangement of seminiferous tubular architecture and degeneration of the Leydig cells in the testis and elevations in the concentrations in the plasma of LH and FSH (p<0.01), commencing at a dose of 5 mg, with the greatest effects at a dose of 20 mg. No significant alterations in these factors occurred at the dose of 2.5 mg in comparison to the control that received only the vehicle. There was no significant alteration in the plasma concentrations of prolactin (p>0.05), corticosterone (p>0.05) or fasting blood glucose or in the rectal temperature (p>0.05) at any of the doses relative to the control group, suggesting that this chemosterilizing procedure did not exert any chronic stress on the experimental animals. From these observations, it may be suggested that 5 mg should be considered as the minimum dose, and 10 mg or 20 mg as the optimum dose, whereas 2.5 mg was ineffective for induction of chemosterilization. There would seem to be little point in using more than 20 mg of calcium chloride for this purpose. Intratesticular injection of calcium chloride at an effective dose may be considered as an alternative to surgical castration.
Similar content being viewed by others
REFERENCES
Aitken, R.J., 1994. A free radical theory of male infertility. Reproduction Fertility and Development, 6, 19-24
Aitken, R.J. and West, K.M., 1990. Analysis of the relationship between ROS production and leucocyte infiltration in fraction of human semen separated on percoll gradients. International Journal of Andrology, 13, 433-451
Albers, G.W. and Theilen, G.H., 1985. Calcium chloride for treatment of subcutaneous lipomas in dogs. Journal of the American Veterinary Medical Association, 186, 492-494
Alvarez, J.G. and Story, B.T., 1984. Assessment of cell damage caused by spontaneous lipid peroxidation in rabbit spermatozoa. Biology of Reproduction, 30, 323-331
Aravinda, S., Gopalakrishnan, B. and Shaha, C., 1996. Testicular GST in germ cell proliferation. In: J. Sengupta and D. Ghosh (eds), Cellular and Molecular Signaling in Reproduction, (New Age International Publishers, New Delhi), 1-10
Barkley, M.S. and Goldmass, B.D., 1977. A quantitative study of serum testosterone. Sex accessory organ growth and the development of intermale aggression in the mouse. Hormones and Behavior, 8, 208-218
Bertlett, J.M.S., Kerr, J.B. and Sharpe, R.M., 1986. The effect of selective destruction and regeneration of rat Leydig cells on the intratesticular distribution of testosterone and morphology of the seminiferous epithelium. Journal of Andrology, 7, 240-253
Buckingham, J.C., Christain, H.C., Gillies, G.E., Philip, J.G. and Taylor, A.D., 1996. The hypothalamo-pituitary-adrenal immune axis. In: J.A. Marsh and M.D. Kandel (eds), The Physiology of Immunity, (CRC Press, New York), 331-354
Chainy, G.B.N., Samantaray, S. and Samanta, L., 1997. Testosterone induced changes in testicular antioxidant system. Andrologia, 29, 343-350
Chowdhury, M., Kabir, S.N., Pal, A.K. and Pakrashi, A., 1983. Modulation of lutinizing hormone receptors: an effect of an inhibitor of prolactin secretion, p-cumeric acid. Journal of Endocrinology, 98, 307-311
Coffey, D.S., 1988. Androgen action and the sex accessory tissues. In: E. Knobil and J. Neill (eds), Physiology of Reproduction, (Raven Press, New York), 1081-1119
Cohen, R.D.H., King, B.D., Thomas, L.R. and Janzen, E.D., 1990. Efficacy and stress of chemical versus surgical castration of cattle. Canadian Journal of Animal Science, 70, 1063-1072
Damassa, D.A., Kobashigawa, D., Smith, E.R. and Davidson, J.M., 1976. Negative feedback control of LH by testosterone: a quantitative study in male rats. Endocrinology, 99, 736-742
Das, D., 1998. Analysis of variance. In: D. Das and A. Das (eds), Statistics in Biology and Psychology, 3rd edn, (Academic Publishers, Calcutta), 250-282
Das, D., 2000. Hormones and neurotransmitters. In: D. Das (ed.), Biochemistry, 10th edn, (Academic Publishers, Calcutta), 595-600
Dhar, J.D. and Setty, B.S., 1990. Effect of a nonsteroidal antiandrogen, anandron, on the reproductive system and fertility in male rats. Contraception, 42, 121-128
Dixit, V.P., Lohiya, N.K., Arya, M. and Argrawal, M., 1973. Chemical sterilization of male dogs after a single testicular injection of ‘Danazol’. Folia Biologica, 23, 305-310
Dowsett, K.F., Pattie, W.A. and Knott, L.M., 1991. A preliminary study of immunological castration in colts. Journal of Reproduction and Fertility, Supplement, 44, 183-190
Dube, D., Assaf, A. and Pelletier, G., 1987. Morphological study of the effects of a GnRH agonist on the canine testis after 4 months of treatment and recovery. Acta Endocrinologia (Copenhagen), 116, 413-417
Foote, R.H., Schermerhorn, E.C. and Simikili, M.E., 1986. Measurement of semen quality, fertility and reproductive hormones to assess dibromochloropropane (DBCP) effects in live rabbits. Fundamental Applied Toxicology, 6, 628-637
Fordyce, G., Hodge, P.B., Beaman, N.J., Leing, H.R., Compro, C. and Shepherd, R.K., 1989. An evaluation of calf castration by intratesticular injection of a lactic acid solution. Assiut Veterinary Medical Journal, 66, 272-276
Georgiou, M., Parkins, L.M. and Payne, A.H., 1987. Steroid synthesis-dependent, oxygen-mediated damage of mitochondrial and microsomal cytochrome P-450 enzymes in rat Leydig cell cultures. Endocrinology, 121, 1390-1399
Ghosh, S.P., Chatterjee, T.K. and Ghosh, J.J., 1983. Synergistic role of prolactin and testosterone in the regulation of acid phosphatase activity and isoenzyme pattern in accessory sex organs of adult male rats. Journal of Reproduction and Fertility, 67, 235-241
Ghosh, S.P., Chowdhury, A., Biswas, N.M. and Ghosh, P.K., 1990. Effect of lithium chloride on testicular steroidogenic and gametogenic functions in male albino rats. Life Sciences, 46, 127-137
Glic, D., Dorothy, Van R. and Lavine, S., 1964. Fluorometric determination of corticosterone and cortisol in 0.02–0.05 milliliters or submilligram samples of adrenal tissue. Endocrinology, 74, 653-657
Gonzalez, A., Allen, A.F. and Post, A.K., 1989. Immunological approaches to contraception in dogs. Journal of Reproduction and Fertility, Supplement, 39, 189-198
Greenwood, F.O., Hunter, W.M. and Glover, J.S., 1963. The preparation of 131I labeled human growth hormone of high specific activity. Biochemistry Journal, 89, 114-123
Gunn, S.A. and Gould, T.C., 1970. Cadmium and other mineral elements. In: A.D. Johnson, W.R. Gomes and N.L. Van Demark (eds), The Testis, vol III, (Academic Press, New York), 377-481
Gutteridge, J.M.C., 1995. Lipid peroxidation and antioxidants as bio markers of tissue damage. Clinical Chemistry, 41, 1812-1828
Gutteridge, J.M.C. and Halliwell, B., 1990. The measurement and mechanism of lipid peroxidation in biological systems. Trends in Biochemical Science, 15, 129-135
Guyton, A.C. and Hall, J.E., 1996. The Adrenocortical Hormones. Textbook of Medical Physiology, 8th edn, (W.B. Saunders, Philadelphia), 963-964
Habig, W.H., Pabst, M.J. and Jocoby, W.B., 1974. Glutathione S-transferase: the first step in mercapturic acid formation. Journal of Biological Chemistry, 249, 7130-7139
Halliwell, B. and Gutteridge, J.M.C., 1985. Free Radicals in Biology and Medicine, (Clarendon Press, Oxford)
Hatayama, I., Satoh, K. and Sato, K., 1986. Development and hormonal regulation of the major form of hepatitis glutathione S-transferase in male mice. Biochemical and Biophysical Research Communications, 140, 581-588
Heath, E. and Arowolo, R., 1987. The early histopathologic effects of intratesticular injection with hyperosmolar glycerol, glucose or NaCl solutions. Andrologia, 19, 654-661
Immegart, H.M. and Threlfall, W.R., 2000. Evaluation of intratesticular injection of glycerol for non surgical sterilization of dogs. American Journal of Veterinary Research, 61, 544-549
Ishii-Ohba, H., Saiki, N., Inano, H. and Takemori, B., 1986. Purification and properties of testicular 3β-hydroxysteroid dehydrogenase and 5-ene-4-ene-isomerase. Journal of Steroid Biochemistry, 25, 255-260
Jacobs, L.S., 1979. Prolactin. In: B.M. Jaffe and H.R. Behrman (eds), Methods of Hormone Radioimmunoassay, (Academic Press, New York), 199-222
Jarabak, J., Adams, J.A., Williams-Ashman, H.G. and Talalay, P., 1962. Purification of a 17β-hydroxysteroid dehydrogenase of human placenta and studies on its transhydrogenase function. Journal of Biological Chemistry, 237, 345-357
Jegou, B. and Sharpe, R.M., 1993. Paracrine mechanism in testicular control. In: D. Dekrester (ed.), Molecular Biology of Male Reproduction, (Academic Press, London), 271-295
Johnson, A.D., 1977. The influence of cadmium on the testis. In: A.D. Johnson and W.R. Gomes (eds), The Testis, vol. IV, (Academic Press, New York), 1191-2001
Johnson, L. and Thomson, D.L. Jr, 1987. Effect of seasonal variation in Leydig cell number on the volume of smooth endoplasmic reticulum in Leydig cells and intratesticular testosterone content in stallions. Journal of Reproduction and Fertility, 81, 227-231
Kakkar, P., Mehrotra, S. and Viswanthan, P.N., 1992. Interrelation of active oxygen species, membrane damage and altered calcium functions. Molecular Cellular Biochemistry, 111, 11-15
Kar, A.B., 1961. Chemical sterilization of male rhesus monkeys. Endocrinology, 69, 1116-1119
Kar, A.B., Kamboj, V.P. and Goswami, A., 1965. Sterilization of male rhesus monkeys by iron salts. Journal of Reproduction and Fertility, 9, 115-117
Kerr, J.B. and Sharpe, R.M., 1986. Effects and interaction of LH and LHRH agonist on testicular morphology and function in hypophysectomized rats. Journal of Reproduction and Fertility, 76, 175-192
Kerr, J.B., Millar, M., Maddocks, S. and Sharpe, R.M., 1993. Stage dependent changes in spermatogenesis and Sertoli cells in relation to the onset of spermatogenic failure following withdrawal of testosterone. Anatomical Record, 235, 547-559
Knigge, U., Matzen, S. and Warberg, J., 1988. Histaminergic mediation of stress induced release of PRL in male rats. Neuroendocrinology, 47, 68-74
Koger, L.M., 1976. Calcium chloride for the prevention of horn bud growth in calves. Journal of the American Veterinary Medical Association, 169, 219-221
Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J., 1951. Protein measurement with folin-phenol reagent. Journal of Biological Chemistry, 193, 265-275
Matsumoto, A.M., 1988. Is high dosage of testosterone an effective male contraceptive agent? Fertility and Sterility, 50, 324-328
McGinnis, K.M., Wangi, K.K. and Genegy, M.E., 1999. Alterations of extra cellular calcium elicit selective modes of cell death and protease activation in SH-SY5Y human neuroblastoma cells. Journal of Neurochemistry, 72, 1853-1863
Mitra, B. and Samanta, P.K., 2000. Testicular degeneration of scrub bulls by calcium chloride. Indian Journal of Veterinary Surgery, 21, 37-38
Munck, A., Guyre, P. and Holbrook, N., 1984. Physiological function of glucocorticoids during stress and their relation to pharmacological actions. Endocrine Review, 5, 25-49
NIHFW, 2001. Protocol for hormone assay by ELISA. In: Orientation Training Course on Research Methodology in Reproductive Biomedicine, (Department of Reproductive Biomedicine, National Institute of Health and Family Welfare, Government of India, New Delhi), 55-58
Naz, R.K. and Talwar, G.P., 1981. Immunological sterilization of male dogs by BCG. International Journal of Andrology, 4, 111-128
NIH, 1985. NIH Guide for the Care and Use of Laboratory Animals. NIH Publication No. 85-23, Revised 1985, (US Department of Health, Education and Welfare, National Institute of Health, Bethesda, MD)
Ohba, H., Inano, H. and Tamaoki, B., 1982. Kinetic mechanism of porcine testicular 17β-hydroxysteroid dehydrogenase. Journal of Steroid Biochemistry, 17, 381-389
Ohkawa, H., Ohishi, N. and Yagi, K., 1979. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry, 95, 351-358
Paoletti, F. and Macoli, A., 1990. Determination of superoxide dismutase activity by a purely chemical system based on NAD(P)H oxidation. Methods in Enzymology, vol. 186, (Academic Press, New York), 209-220
Parizek, J., 1960. Sterilization of the male by cadmium salts. Journal of Reproduction and Fertility, 1, 294-309
Peltola, V., Huhtaniemi, I. and Ahotupa, M., 1992. Antioxidant enzyme activity in the maturing rat testis. Journal of Andrology, 13, 450-455
Peltola, V., Huhtaniemi, I., Metsa-ketla, T. and Ahotupa, M., 1996. Induction of lipid peroxidation during steroidogensis in rat testis. Endocrinology, 137, 105-112
Plant, T.M., 1982. Effects of orchidectomy and testosterone replacement treatment on pulsatile lutinizing hormone secretion in the adult Rhesus monkey (Macaca mulatta). Endocrinology, 110, 1905-1913
Rao, A.V.S.K. and Shaha, C., 2000. Role of glutathione S-transferase in oxidative stress-induced male germ cell apoptosis. Free Radical Biology and Medicine, 29, 1015-1027
Rivier, C., Rivier, J. and Vale, W., 1979. Chronic effects of [D-Trp6,Pro9-Net] lutinizing hormone releasing factor on reproductive process in the male rat. Endocrinology, 105, 1191-1201
Sadasivam, S. and Manickam, A., 1996. Peroxidase. In: S. Sadasivam and A. Manickam (eds), Methods in Biochemistry, (New Age International Publishers, New Delhi), 108-110
Samanta, P.K., 1998. Chemosterilization of stray dogs. Indian Journal of Animal Health, 37, 61-62
Samanta, L., Roy, A. and Chainy, G.B.N., 1999. Changes in rat testicular antioxidant defense profile as a function of age and its impairment by hexachlorocyclohexane during critical stages of maturation. Andrologia, 31, 83-90
Seltzer, A.M., Donoso, A.O. and Podesta, E., 1986. Restraint stress stimulation of PRL and ACTH secretion: role of brain histamine. Physiology and Behaviour, 36, 251-255
Sharpe, R.M., Donachie, K. and Cooper, I., 1988. Re-evaluation of the intratesticular level of testosterone required for quantitative maintenance of spermatogenesis in the rat. Journal of Endocrinology, 117, 19-26
Sharpe, R.M., Maddocks, S., Millar, M., Saunders, P.T.K., Kerr, J.B. and McKinnell, C., 1992. Testosterone and spermatogenesis: identification of stage dependent, androgen-regulated proteins secreted by adult rat seminiferous tubules. Journal of Andrology, 13, 172-184
Sikka, S.C., 2001. Relative impact of oxidative stress on male reproductive function. Current Medicinal Chemistry, 8, 851-862
Silber, R.H., 1966. Fluorimetric analysis of corticoids. In: D. Glick (ed.), Methods in Biochemical Analysis, (Interscience Publishers, New York), 63
Slater, T.I., 1984. Overview of methods used for detecting lipid peroxidation. Methods in Enzymology, vol. 105, (Academic Press, New York), 283-293
Steinberger, E., 1971. Hormonal control of mammalian spermatogenesis. Physiology Review, 51, 1-22
Steinberger, E., 1975. Hormonal regulation of the seminiferous tubule function. In: F.S. French, V. Hansson, E. Ritzen and S.N. Nayfeh (eds), Hormonal Regulation of Spermatogenesis, (Plenum Press, New York), 337-352
Swerdloff, R.S., Wang, C. and Bhasin, S., 1992. Developments in the control of testicular function. Baillière's Clinical Endocrinology and Metabolism, 6, 451-483
Talalay, P., 1962. Hydroxysteroid dehydrogenase. In: S.P. Colowick and N.O. Kaplan (eds), Methods in Enzymology, vol. 5, (Academic Press, New York), 512-516
Tillbrook, A.J. and Clarke, I.J., 1995. Negative feedback regulation of the secretion and actions of GnRH in male ruminants. Journal of Reproduction and Fertility, Supplement, 49, 297-306
Tillbrook, A.J. and Clarke, I.J., 2001. Negative feedback regulation of the secretion and actions of gonadotrophin releasing hormone in males. Biology of Reproduction, 64, 735-742
Trembley, Y. and Belanger, A., 1984. Reversible inhibition of gonadal functions by a potent gonadotrophin releasing hormone agonist in adult dog. Contraception, 30, 483-497
Watanabe, H., Kobayashi, A., Yamamoto, T., Suzuki, S., Hayashi, H. and Yamazaki, N., 1990. Alteration of human erythrocyte membrane fluidity by oxygen-derived free radicals and calcium. Free Radical Biology and Medicine, 8, 507-514
Wiebe, J.P., Barr, K.J. and Buckingham, K.D., 1989. Sustained azospermia in the squirrel monkey, Saimiri sciureus, resulting from a single intratesticular glycerol injection. Contraception, 39, 447-457
Willett, L.B. and Erb, R.E., 1972. Short term changes in plasma corticoids in dairy cattle. Journal of Animal Science, 34, 103-111
Wu, F.C.W. and Aitken, R.J., 1989. Suppression of sperm function by depot-medroxyprogesterone acetate and testosterone enanthate in steroid male contraception. Fertility and Sterility, 29, 443-465
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Jana, K., Samanta, P. & Ghosh, D. Dose-dependent Response to an Intratesticular Injection of Calcium Chloride for Induction of Chemosterilization in Adult Albino Rats. Vet Res Commun 26, 651–673 (2002). https://doi.org/10.1023/A:1020976905746
Issue Date:
DOI: https://doi.org/10.1023/A:1020976905746