The objective of this proposal is to investigate the mechanism by which testicular androgen induces the fetal mouse genital tract differentiation. During the last few, years, we made some observations which suggest that fetal androgen after binding to its receptor induces the synthesis of phospholipase stimulatory protein(s) resulting in stimulation of phospholipase A2 (PLA2). This, in turn, induces the arachidonic acid release leading to increased synthesis of prostaglandin E2 (PGE2). Consequently, masculine differentiation is induced. We plan to investigate this hypothesis further by determining specific roles of different intermediates of the above-mentioned pathway, namely, PGE2, PLC and phospholipase A2-stimulatory proteins (PLSP). Thus, we propose (1) to investigate whether a direct exposure of a PG depriving agent, namely, indomethacin, in an in vitro organ culture system results in the complete prevention of the androgen-induced masculinization i.e. disruption of the differentiation of internal genitalia, which was not observed with the indirect in vivo exposure; (2) to investigate whether PGE2 modulates ornithine decarboxylase to induce the differentiation; (3) to determine whether induction of PLC during the differentiation results in an induction of diacylglycerol and inositol triphosphate, the cell proliferating system; (4) to identify and characterize the PLA2-stimulatory and masculinizing activities of male PLSP using polydonal and monoclonal antibodies prepared against the protein(s) with these activities; (5) to investigate the difference between male and female PLSP, and (6) to determine the synthesis and localization of PLA2-stimulatory and masculinizing proteins during the genital tract differentiation of males and females.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
2R01HD022781-05A1
Application #
3322627
Study Section
Biochemical Endocrinology Study Section (BCE)
Project Start
1987-04-01
Project End
1994-07-31
Budget Start
1991-08-01
Budget End
1992-07-31
Support Year
5
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Children's Hosp Pittsburgh/Upmc Health Sys
Department
Type
DUNS #
044304145
City
Pittsburgh
State
PA
Country
United States
Zip Code
15224
Nguyen, A P; Chandorkar, A; Gupta, C (1996) The role of growth hormone in fetal mouse reproductive tract differentiation. Endocrinology 137:3659-66
Gupta, C (1994) The 72-kilodalton protein of the male reproductive tract is differentially expressed and developmentally regulated during Wolffian duct differentiation of the fetal mouse. Biol Reprod 51:283-9
Gupta, C (1993) Further characterization of the testosterone inducible protein fraction from the mouse fetal male reproductive tract inducing masculine differentiation in vitro. J Steroid Biochem Mol Biol 45:375-81
Gupta, C; Bentlejewski, C (1993) Immunological identification of the protein-producing masculine differentiation of the Wolffian duct in the fetal mouse: western blot analysis and determination of the biological activity. Endocrinology 133:1341-6
Gupta, C; Bentlejewski, C A (1992) Role of prostaglandins in the testosterone-dependent wolffian duct differentiation of the fetal mouse. Biol Reprod 47:1151-60
Gupta, C; Braun, A (1990) Identification and partial purification of embryonic mouse genital protein(s) stimulating phospholipase-A2 and inducing masculinization in vitro. Endocrinology 126:341-8
Gupta, C; Wharton, V; Ellis, D (1990) Anti-masculinizing action of estradiol and cyproterone acetate: regulation of a protein fraction with phospholipase-A2 stimulatory and masculinizing activities. J Steroid Biochem Mol Biol 37:661-7
Gupta, C (1989) Prostaglandins masculinize the mouse genital tract. Endocrinology 124:1781-7
Gupta, C (1989) The role of prostaglandins in masculine differentiation: modulation of prostaglandin levels in the differentiating genital tract of the fetal mouse. Endocrinology 124:129-33
Gupta, C (1988) Activation of phospholipases during masculine differentiation of embryonic genitalia. Proc Soc Exp Biol Med 188:489-94