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Stage-Specific Nuclear Expression of NF-B in Mammalian Testis

Department of Cell Biology and Physiology University of Pittsburgh Pittsburgh, Pennsylvania 15261

The Rel/nuclear factor (NF)-B family of transcription factors are important intracellular conveyors of extracellular signals in a number of systems. However, little is known of their roles in the specialized, hormonally regulated environment of the mammalian testis. In this study NF-B p50 and p65 proteins were found to be constitutively present and active in the nucleus of Sertoli cells cultured from rat testis. In vivo, NF-B proteins are present in the nucleus of Sertoli cells during all 14 (I–XIV) cyclical stages of spermatogenesis; however, nuclear NF-B expression was elevated in stage XIV and remained high in stages I–VII. In contrast, NF-B p50 and p65 subunits are transiently expressed in the nuclei of germ cells with peak levels found in pachytene spermatocytes during stages VII–XI and lower levels in stage I-VII spermatids. Tumor necrosis factor-, which is produced by round spermatids in the testis, increased nuclear NF-B binding activity when added to Sertoli cells. Stimulation of Sertoli cells with activators of the cAMP-protein kinase A (PKA) signaling pathway such as forskolin or FSH also increased NF-B DNA binding activity. Consistent with the cellular localization studies, NF-B was found to be activated as high basal levels of NF-B-stimulated reporter gene expression were detected in transient transfection studies of Sertoli cells. Addition of tumor necrosis factor- to Sertoli cells further stimulated B enhancer-mediated transcription. These findings suggest that NF-B proteins are stage specifically localized to Sertoli cell and spermatocyte nuclei and may play a role in the regulation of stage-specific gene expression during the process of spermatogenesis.

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