The long term objective of this project is to determine the influence of genetic control on steroidogenesis. The studies proposed in this application are designed to examine the relationships among the structural and regulatory gene(s) that influence the expression of the enzymes involved in testicular steroid hormone biosynthesis. Specifically, it is proposed to determine the chromosomal location of genes determining quantitative and structural variation of the following steroidogenic enzymes: 3 beta-hydroxysteroid dehydrogenase-isomerase (3 beta-HSD), cholesterol side-chain cleavage (P-450scc) and 17 alpha-hydroxylase/C17-20 lyase (P-450-17 alpha). Whole testicular preparations, purified Leydig cells or adrenal glands from numerous strains of inbred mice will be surveyed for quantitative and qualitative differences of each of the enzymes. Recombitant inbred (RI) lines, derived from progenitor strains which show the greatest interstrain differences, will be used to map the chromosomal location of the structural and regulatory gene(s) for each enzyme. Quantitative differences in 3 beta-HSD will be determined by measuring enzyme activity; structural variants of this enzyme will be detected by measuring rate of heat inactivation (thermostability). Quantitative differences in P-450scc and P-450-17 alpha will be determined in lysates of purified Leydig cells or of adrenal glands using specific rabbit antisera generated against these enzymes plus 125I-labeled second antibody and analysis by immunoblotting. Structural variants of P-450scc and P-450-17 alpha will be detected by analyzing for variations in restriction fragment length of genomic DNA, from different inbred strains of mice, through hybridization with cDNA probes specific for P-450scc and P-450-17 alpha. The availability of specific antisers to the P-450scc and P-450-17 alpha and the cDNA clones specific for each of these enzymes provide sensitive reagents for the detection of quantitative and qualitative variants, respectively. Information gained from the proposed studies will provide a better understanding of the normal physiologic function of the Leydig cell as well as explanations for pathologic states due to genetic defects in steroid biosynthesis enzymes in testes and adrenal glands.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD017916-07
Application #
3314928
Study Section
Endocrinology Study Section (END)
Project Start
1983-09-01
Project End
1991-08-31
Budget Start
1989-09-01
Budget End
1990-08-31
Support Year
7
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Payne, A H; Abbaszade, I G; Clarke, T R et al. (1997) The multiple murine 3 beta-hydroxysteroid dehydrogenase isoforms: structure, function, and tissue- and developmentally specific expression. Steroids 62:169-75
Burgos-Trinidad, M; Youngblood, G L; Maroto, M R et al. (1997) Repression of cAMP-induced expression of the mouse P450 17 alpha-hydroxylase/C17-20 lyase gene (Cyp17) by androgens. Mol Endocrinol 11:87-96
Abbaszade, I G; Arensburg, J; Park, C H et al. (1997) Isolation of a new mouse 3beta-hydroxysteroid dehydrogenase isoform, 3beta-HSD VI, expressed during early pregnancy. Endocrinology 138:1392-9
Clarke, T R; Bain, P A; Burmeister, M et al. (1996) Isolation and characterization of several members of the murine Hsd3b gene family. DNA Cell Biol 15:387-99
Park, C H; Abbaszade, I G; Payne, A H (1996) Expression of multiple forms of 3 beta-hydroxysteroid dehydrogenase in the mouse liver during fetal and postnatal development. Mol Cell Endocrinol 116:157-64
Payne, A H; Youngblood, G L (1995) Regulation of expression of steroidogenic enzymes in Leydig cells. Biol Reprod 52:217-25
Payne, A H; Clarke, T R; Bain, P A (1995) The murine 3 beta-hydroxysteroid dehydrogenase multigene family: structure, function and tissue-specific expression. J Steroid Biochem Mol Biol 53:111-8
Abbaszade, I G; Clarke, T R; Park, C H et al. (1995) The mouse 3 beta-hydroxysteroid dehydrogenase multigene family includes two functionally distinct groups of proteins. Mol Endocrinol 9:1214-22
Greco, T L; Payne, A H (1994) Ontogeny of expression of the genes for steroidogenic enzymes P450 side-chain cleavage, 3 beta-hydroxysteroid dehydrogenase, P450 17 alpha-hydroxylase/C17-20 lyase, and P450 aromatase in fetal mouse gonads. Endocrinology 135:262-8
Bain, P A; Meisler, M H; Taylor, B A et al. (1993) The genes encoding gonadal and nongonadal forms of 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase are closely linked on mouse chromosome 3. Genomics 16:219-23

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