Sperm maturation probably requires interaction of sperm with specific secretory products of the epididymal epithelium. Sperm maturation occurs in the proximal epididymis if locally produced dihydrotestosterone is available to stimulate secretion by the epithelial cells. Maintenance of the enzyme 5Alpha-reductase may be dependent on continuous availability of component(s), possibly androgen binding protein, in rete testis fluid. In preparation for testing the hypotheses that a factor(s) in rete testis fluid controls activity of epididymal 5Alpha-reductase and hence epididymal secretion and that secretion of principal cells from specific regions of the epididymis bind to immature sperm and alter their function, we have developed a procedure to culture principal cells from specific regions of the ram epididymis. Ovine rete testis fluid and immature sperm will be obtained by cannulating the rete testis. Two potential control points for contraceptive action will be examined by studies aimed at (a) identification of the regulatory component(s) in rete testis fluid and (b) establishment of the role of eipididymal secretory products in sperm maturation. The role of rete testis fluid in controlling epididymal function will be studied by culturing highly purified preparations of ram principal cells, from specific regions, of the epididymis, in media containing 20 or 100% concentration of ram rete testis fluid or fractions thereof. Cell response will be measured by activity of 5Alpha-reductase, cell ultrastructure, and qualitative analyses of secreted proteins. The role of the hormonal milieu (androgen, progesterone, estradiol and aldosterone) on characteristics of the cells and their secretions will be evaluated similarly. Differences among secretions from different regions of the epididymis will be examined. Finally, the binding of secreted proteins by immature (testicular) ram sperm will be examined and the responses of the sperm to bound secretory protein will be evaluated. Success in manipulation of either control point for opididymal function could allow inhibition of sperm maturation oand reversible contraception.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD014501-06
Application #
3312644
Study Section
Reproductive Biology Study Section (REB)
Project Start
1980-03-01
Project End
1986-11-30
Budget Start
1985-03-01
Budget End
1986-11-30
Support Year
6
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Colorado State University-Fort Collins
Department
Type
Schools of Veterinary Medicine
DUNS #
112617480
City
Fort Collins
State
CO
Country
United States
Zip Code
80523
Veeramachaneni, D N; Amann, R P (1991) Endocytosis of androgen-binding protein, clusterin, and transferrin in the efferent ducts and epididymis of the ram. J Androl 12:288-94
Veeramachaneni, D N; Amann, R P (1990) Oxytocin in the ovine ductuli efferentes and caput epididymidis: immunolocalization and endocytosis from the luminal fluid. Endocrinology 126:1156-64
Veeramachaneni, D N; Amann, R P; Palmer, J S et al. (1990) Proteins in luminal fluid of the ram excurrent ducts: changes in composition and evidence for differential endocytosis. J Androl 11:140-54
Marengo, S R; Amann, R P (1990) Morphological features of principal cells in the ovine epididymis: a quantitative and qualitative study. Biol Reprod 42:167-79
Veeramachaneni, D N; Schanbacher, B D; Amann, R P (1989) Immunolocalization and concentrations of inhibin alpha in the ovine testis and excurrent duct system. Biol Reprod 41:499-503
Tekpetey, F R; Veeramachaneni, D N; Amann, R P (1989) Localization of androgen receptors in ram epididymal principal cells. J Reprod Fertil 87:311-9
Tekpetey, F R; Amann, R P (1988) Regional and seasonal differences in concentrations of androgen and estrogen receptors in ram epididymal tissue. Biol Reprod 38:1051-60
Tekpetey, F R; Amann, R P (1988) Effects of exogenous melatonin prior to the breeding season on testis weight and epididymal androgen receptors in rams. Domest Anim Endocrinol 5:257-64
Knickerbocker, J J; Sawyer, H R; Amann, R P et al. (1988) Evidence for the presence of oxytocin in the ovine epididymis. Biol Reprod 39:391-7
Amann, R P; Marengo, S R; Brown, D V (1987) Steroidogenesis and testosterone metabolism in cultured principal cells from the ram epididymis. J Androl 8:238-46

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