The long term objective of this research is the elucidation of the mechanisms which are responsible for the development of sperm fertilizing capacity in the mammalian epididymis. As spermatozoa pass through the epididymis, they are sequentially exposed to varying epididymal environments created by the secretion and absorption of epididymal cells. The working hypothesis is that the maturation of the sperm plasma membrane necessary for sperm-egg interaction is dependent upon the sequential interaction with hormonally controlled epididymal secretory proteins. The following procedures have been developed: 1) maintainance in vitro of epididymal explants where the interaction between all the cells of a region and their 3-dimensional structural relationship are preserved, and 2) isolation and culture of viable epithelial and peritubular epididymal cells. The specific objectives of this proposal are 1) to establish the optimal conditions for the study of protein synthesis and secretion of epididymal cells. This will be studied, after incorporation of labeled sugars and amino acids, by light and electron microscope autoradiography, SDS-PAGE electrophoresis, and immunoprecipitation; 2) to study the hormonal control of protein and glycoprotein synthesis and secretion; 3) to determine the affinity of secreted proteins and glycoprotein for epididymal spermatozoa and their effect first on sperm-egg binding and then on sperm fertilizing ability; 4) to determine the regional differences in epididymal secretion and secretion-sperm interactions; 5) to determine whether epididymal cells secrete glycosyltransferases and glycosidases which can modify the sperm surface; 6) to study epididymal secretion and secretion-sperm interaction in mutants with normal sperm production, sperm morphology, sperm motility, and low fertility; 7) to study epididymal protein synthesis and secretion during prepuberal development. Rabbits, rats, and mice will be used. These studies should provide important insights into the problem of male fertility.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
2R37HD003820-18
Application #
3484915
Study Section
Reproductive Biology Study Section (REB)
Project Start
1968-12-01
Project End
1991-11-30
Budget Start
1986-12-01
Budget End
1987-11-30
Support Year
18
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37203
Li, Shengxu; Chen, Wei; Srinivasan, Sathanur R et al. (2004) Childhood blood pressure as a predictor of arterial stiffness in young adults: the bogalusa heart study. Hypertension 43:541-6
Lareyre, J J; Winfrey, V P; Kasper, S et al. (2001) Gene duplication gives rise to a new 17-kilodalton lipocalin that shows epididymal region-specific expression and testicular factor(s) regulation. Endocrinology 142:1296-308
Chayko, C A; Orgebin-Crist, M C (2000) Targeted disruption of the cation-dependent or cation-independent mannose 6-phosphate receptor does not decrease the content of acid glycosidases in the acrosome. J Androl 21:944-53
Chayko, C A; Orgebin-Crist, M C; Skudlarek, M D et al. (2000) Biosynthesis, processing, and subcellular localization of rat spermbeta-D-galactosidase. Biol Reprod 63:688-96
Lareyre, J J; Reid, K; Nelson, C et al. (2000) Characterization of an androgen-specific response region within the 5' flanking region of the murine epididymal retinoic acid binding protein gene. Biol Reprod 63:1881-92
Lareyre, J J; Thomas, T Z; Zheng, W L et al. (1999) A 5-kilobase pair promoter fragment of the murine epididymal retinoic acid-binding protein gene drives the tissue-specific, cell-specific, and androgen-regulated expression of a foreign gene in the epididymis of transgenic mice. J Biol Chem 274:8282-90
Lareyre, J J; Zheng, W L; Zhao, G Q et al. (1998) Molecular cloning and hormonal regulation of a murine epididymal retinoic acid-binding protein messenger ribonucleic acid. Endocrinology 139:2971-81
Lareyre, J J; Mattei, M G; Kasper, S et al. (1998) Genomic organization and chromosomal localization of the murine epididymal retinoic acid-binding protein (mE-RABP) gene. Mol Reprod Dev 50:387-95
Abou-Haila, A; Orgebin-Crist, M C; Skudlarek, M D et al. (1998) Identification and androgen regulation of egasyn in the mouse epididymis. Biochim Biophys Acta 1401:177-86
Tulsiani, D R; Orgebin-Crist, M C; Skudlarek, M D (1998) Role of luminal fluid glycosyltransferases and glycosidases in the modification of rat sperm plasma membrane glycoproteins during epididymal maturation. J Reprod Fertil Suppl 53:85-97

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