The two primary objectives currently pursued are to elucidate the effects of follicle stimulating hormone (FSH) on the polyamine (PA) biochemistry of the Sertoli cell (SC) and to determine whether or not FSH receptors (FSHR) may be implicated in infertility of an autoallergic etiology utilizing the Granulosa cell (GC) as an experimental model. The 1st project focuses on 3 specific areas: 1) The role of FSH in the induction of ornithine decarboxylase activity and mRNA accumulation and its effects on PA biosynthesis in SC, 2) The role of transglutamination and the effects of transglutaminase (TGase) inhibition on SC function, 3) Characterization of a spermine binding site in SC. The effects of media constituents on ODC induction and PA biosynthesis as well as the effects of PA depletion (inhibition of ODC by difluoromethyl ornithine and spermine synthase by s-adenosyl-1,8-diamine-3-thiooctane) on these processes as well as steroidogenesis and the cellular processing of FSH by SC will be investigated. TGase activity in SC will be studied with regard to its induction by serum factors and its substrate specificity. The spermine binding site binding affinity, specificity and chemical properties will be determined. The presence of a spermine binding protein on SC suggests a possible mechanism for cell to cell communication within the tubule lumen or between testicular compartments. The working hypothesis of the 2nd project suggests that antibody inactivation of the FSHR and/or membrane associated entities results in gonadal failure. Inactivation may occur by 1) the FSHR becoming antigenic by virtue of its association with FSH which was involved in a primary antibody response (heterogenization) 2) a secondary response antibody is produced which recognized the first antibody which was formed to FSH (anti-idiotype formation). In the latter case, antibodies directed against the paratope of the anti-FSH antibodies would interact directly with the FSH binding region of R. In order to directly test the hypothesis, the effect of active immunization with purified antibody to FSH will be studied. Antisera with a positive titer for anti-idiotype will be tested for the presence of anti-R antibodies. It will be necessary to initially characterize the FSHR in GC and to prepare an isotopically labeled FSHR for later use in immunodiagnostic studies. The long term goals are to establish which specific biochemical processes are essential to FSHR function and modulation of FSH mediated cellular activation and to determine whether or not FSHR may be implicated in infertility of an auto-allergic etiology.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Modified Research Career Development Award (K04)
Project #
5K04HD000682-03
Application #
3073276
Study Section
Biochemical Endocrinology Study Section (BCE)
Project Start
1985-08-01
Project End
1988-02-29
Budget Start
1987-08-01
Budget End
1988-02-29
Support Year
3
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Albany Medical College
Department
Type
Schools of Medicine
DUNS #
City
Albany
State
NY
Country
United States
Zip Code
12208
Weiner, K X; Dias, J A (1993) Regulation of ovarian ornithine decarboxylase activity and its mRNA by gonadotropins in the immature rat. Mol Cell Endocrinol 92:195-9
Weiner, K X; Dias, J A (1992) Developmental regulation of ornithine decarboxylase (ODC) in rat testis: comparison of changes in ODC activity with changes in ODC mRNA levels during testicular maturation. Biol Reprod 46:617-22
Weiner, R S; Dias, J A (1992) Biochemical analyses of proteolytic nicking of the human glycoprotein hormone alpha-subunit and its effect on conformational epitopes. Endocrinology 131:1026-36
Vakharia, D D; Bryant, S H; Dias, J A (1992) Topographic analysis of human follicle-stimulating hormone-beta using anti-peptide antisera. Mol Cell Endocrinol 85:89-97
Weiner, R S; Dias, J A; Andersen, T T (1991) Epitope mapping of human follicle stimulating hormone-alpha using monoclonal antibody 3A identifies a potential receptor binding sequence. Endocrinology 128:1485-95
Krystek Jr, S R; Dias, J A; Andersen, T T (1991) Identification of subunit contact sites on the alpha-subunit of lutropin. Biochemistry 30:1858-64
Vakharia, D D; Dias, J A; Andersen, T T (1991) Determination of subunit contact-associated epitopes of the beta-subunit of human follicle-stimulating hormone. Endocrinology 128:1797-804
Weiner, K X; Dias, J A (1990) Protein synthesis is required for testosterone to decrease ornithine decarboxylase messenger RNA levels in rat Sertoli cells. Mol Endocrinol 4:1791-8
Vakharia, D D; Dias, J A; Thakur, A N et al. (1990) Mapping of an assembled epitope of human follicle-stimulating hormone-beta utilizing monoclonal antibodies, synthetic peptides, and hormone-receptor inhibition. Endocrinology 127:658-66
Weiner, K X; Pentecost, B T; Dias, J A (1990) Testosterone decreases ornithine decarboxylase messenger RNA levels in primary cultures of rat Sertoli cells. Mol Endocrinol 4:1249-56

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