The objective is to understand the role of estrogen receptors in the regulation of uterine growth and function by elucidating the important receptor changes which occur concomitantly or are required for estrogen regulation of gene expression in target cells. This is to be accomplished by studying the steroid hormone-regulated expression of several recently characterized estrogen-dependent secretory rat endometrial proteins, correlated with estrogen-dependent receptor modulation in the same cells, documenting changes in the receptor protein with a library of monoclonal antibodies to the estrogen receptor as well as steroid binding methods. The estrogen-responsive, marker proteins of the rat endometrial epithelium will be isolated, purified and characterized and used for preparation of polyclonal and monoclonal antibodies. These marker protein antibodies will be used to identify the location and possible function of the proteins and to isolate the specific m-RNAs for the proteins. After preparation of c-DNA, detailed studies will characterize the nature of the regulation of the marker at the level of transcription and translation. The c-DNA will eventually be used to screen genomic libraries to identify the gene for the estrogen-dependent markers so that the molecular details of the receptor regulated expression of the marker can provide new insight into the estrogen regulation of gene expression in the uterus. Immunoblots, binding and fractionation studies as well as immunohistochemical assays with the monoclonal antibodies to the receptor will be used to characterize changes in the nature, content and localization of the estrogen receptor in the different types of uterine cells under various hormonal conditions. Immunomethods will be used to study the major receptor events, such as estrogen receptor synthesis, potentiation, activation, processing, recycling, degradation and receptor binding to chromatin and nuclear matrix. These studies can be expected to provide new information on changes in the estrogen receptor, not available simply from assay of estrogen-binding studies, and to improve our understanding of the molecular details of steroid hormone regulation of the individual cell types of the uterus at the molecular level. This new knowledge will be useful for better diagnosis and treatment of steroid hormone-dependent cancers and will provide useful information for the control of normal and abnormal reproductive function.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD015513-09
Application #
3313137
Study Section
Biochemical Endocrinology Study Section (BCE)
Project Start
1981-08-01
Project End
1990-07-31
Budget Start
1989-08-01
Budget End
1990-07-31
Support Year
9
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of Chicago
Department
Type
Schools of Medicine
DUNS #
225410919
City
Chicago
State
IL
Country
United States
Zip Code
60637
Puy, L A; Kuivanen, P C; DeSombre, E R (1993) Immunohistochemical localization of the oestrogen-responsive 110 kDa and 74 kDa polypeptides and complement component C3 in the rat genital tract after oestrogen treatment and during the oestrous cycle. J Reprod Fertil 99:385-94
DeSombre, E R; Hughes, A; Mease, R C et al. (1990) Comparison of the distribution of bromine-77-bromovinyl steroidal and triphenylethylene estrogens in the immature rat. J Nucl Med 31:1534-42
Kuivanen, P C; Capulong, R B; Harkins, R N et al. (1989) The estrogen-responsive 110K and 74K rat uterine secretory proteins are structurally related to complement component C3. Biochem Biophys Res Commun 158:898-905
DeSombre, E R; Mease, R C; Sanghavi, J et al. (1988) Estrogen receptor binding affinity and uterotrophic activity of triphenylhaloethylenes. J Steroid Biochem 29:583-90
DeSombre, E R; Harper, P V; Hughes, A et al. (1988) Bromine-80m radiotoxicity and the potential for estrogen receptor-directed therapy with auger electrons. Cancer Res 48:5805-9
DeSombre, E R; Mease, R C; Hughes, A et al. (1988) Bromine-80m-labeled estrogens: Auger electron-emitting, estrogen receptor-directed ligands with potential for therapy of estrogen receptor-positive cancers. Cancer Res 48:899-906
Mobbs, B G; Johnson, I E; DeSombre, E R et al. (1987) Regulation of estrogen and progestin receptor concentrations in an experimental rat prostatic carcinoma by estrogen, antiestrogen, and progesterone. Cancer Res 47:2645-51
Kuivanen, P C; DeSombre, E R (1985) The effects of sequential administration of 17 beta-estradiol on the synthesis and secretion of specific proteins in the immature rat uterus. J Steroid Biochem 22:439-51