The 72-Kilodalton Protein of the Male Reproductive Tract is Differentially Expressed and Developmentally Regulated during Wolffian Duct Differentiation of the Fetal Mouse1

Abstract

Differentiation of the mammalian reproductive tract into the male phenotype is dependent on the presence of functional testes secreting testosterone during in utero development. Recently, we identified a protein fraction (72 kDa) from the developing reproductive tract of the fetal male mouse that is able to mimic the effect of testosterone in stabilizing the Wolffian duct in vitro, suggesting a role played by this protein in mediating the effect of testosterone during Wolffian duct differentiation. In the present study, we further evaluated the masculinizing role of this protein by determining the specific appearance of the 72-kDa protein in relation to reproductive tract differentiation. Using Western blot and immunohistochemical analysis, we demonstrate that the protein was expressed specifically in the male reproductive tract and not in male non-reproductive organs, namely, lung, liver, or kidney. Within the male reproductive tract, the Wolffian duct derivatives demonstrated a high level of expression of the protein, whereas the urogenital sinus produced only a low level of reaction to this antibody in Western analysis and no reaction in the histochemical analysis. There was a gradual rise in expression of the protein in the male reproductive tract during the critical period of Wolffian duct differentiation. The female reproductive tract produced only a trace immune reaction in Western analysis and no reaction in immunohistochemical analysis. Thus, the results support the hypothesis that the 72-kDa protein plays a role in Wolffian duct differentiation.