Progesterone is a steroid hormone required both for implantation of the fertilized ovum, as well as for the maintenance of pregnancy. The goal of the proposed research is to elucidate the mechanisms whereby the synthesis of progesterone is regulated in the ovary and placenta, organs characterized by exceedingly high rates of secretion of progesterone. Previously we have shown that low-density lipoprotein (LDL)-cholesterol is a major source of precursor for progesterone biosynthesis in these tissues. Furthermore, we have shown in preliminary studies that LDL induces the synthesis of the enzymes involved in the conversion of cholesterol to pregnenolone, the first step in progesterone biosynthesis. Utilizing luteal cells and granulosa cells in monolayer culture, the effect of gonadotropins to induce the synthesis of the enzymes involved in the conversion of cholesterol to pregnenolone will be investigated. Utilizing luteal, granulosa, trophoblastic and choriocarcinoma cells in monolayer culture, the effect of LDL to induce the synthesis of these proteins will be examined. Cells will be radiolabeled with 35S methionine, or else RNA will be extracted from the cells and used to program a cell-free translation system in the presence of [35S]methionine. Enzymes involved in the conversion of cholesterol to pregnenolone will be subjected to immunoprecipitation, separated by SDS-polyacrylamide gel electrophoresis, and visualized by means of autoradiography. Species of cDNA specific for these proteins will be isolated from a cDNA library prepared from bovine adrenocortical poly A RNA. These cDNA probes will be utilized to determine whether treatment of cells with gonadotropins, as well as LDL, leads to an increase in the content of mRNA species specific for these enzymes. In addition to progesterone secretion, the process of in vitro luteinization will be investigated in order to ascertain whether LDL is a major factor regulating this process. The results of these studies will allow us to evaluate the concept that LDL is a major physiological factor regulating progesterone secretion in the ovary and placenta and the process of luteinization in the ovary, and will enable us to explore the cellular and molecular mechanisms of a hitherto unknown action of LDL, namely, to induce the synthesis of the enzymes catalyzing the conversion of cholesterol to progesterone.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD013234-10
Application #
3312143
Study Section
Biochemical Endocrinology Study Section (BCE)
Project Start
1979-07-01
Project End
1989-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
10
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Texas Sw Medical Center Dallas
Department
Type
Schools of Medicine
DUNS #
City
Dallas
State
TX
Country
United States
Zip Code
75390
Liu, Z; Simpson, E R (1999) Molecular mechanism for cooperation between Sp1 and steroidogenic factor-1 (SF-1) to regulate bovine CYP11A gene expression. Mol Cell Endocrinol 153:183-96
Fisher, C R; Graves, K H; Parlow, A F et al. (1998) Characterization of mice deficient in aromatase (ArKO) because of targeted disruption of the cyp19 gene. Proc Natl Acad Sci U S A 95:6965-70
Simpson, E R (1998) Genetic mutations resulting in estrogen insufficiency in the male. Mol Cell Endocrinol 145:55-9
Liu, Z; Simpson, E R (1997) Steroidogenic factor 1 (SF-1) and SP1 are required for regulation of bovine CYP11A gene expression in bovine luteal cells and adrenal Y1 cells. Mol Endocrinol 11:127-37
Borroni, R; Liu, Z; Simpson, E R et al. (1997) A putative binding site for Sp1 is involved in transcriptional regulation of CYP17 gene expression in bovine ovary. Endocrinology 138:2011-20
Simpson, E R; Michael, M D; Agarwal, V R et al. (1997) Cytochromes P450 11: expression of the CYP19 (aromatase) gene: an unusual case of alternative promoter usage. FASEB J 11:29-36
Hinshelwood, M M; Dodson Michael, M; Sun, T et al. (1997) Regulation of aromatase expression in the ovary and placenta: a comparison between two species. J Steroid Biochem Mol Biol 61:399-405
Simpson, E R; Zhao, Y; Agarwal, V R et al. (1997) Aromatase expression in health and disease. Recent Prog Horm Res 52:185-213; discussion 213-4
Michael, M D; Michael, L F; Simpson, E R (1997) A CRE-like sequence that binds CREB and contributes to cAMP-dependent regulation of the proximal promoter of the human aromatase P450 (CYP19) gene. Mol Cell Endocrinol 134:147-56
Conley, A J; Corbin, C J; Hinshelwood, M M et al. (1996) Functional aromatase expression in porcine adrenal gland and testis. Biol Reprod 54:497-505

Showing the most recent 10 out of 81 publications