Apoptosis control in adult testis is crucial to achieve normal spermatogenesis. In this study c-FLIP, an apoptosis-modulating protein, was investigated. In Western blot and immunohistochemical analyses, the 55 KDa c-FLIP long isoform (c- FLIPL) was found to be expressed strongly in spermatocytes and spermatids, at low levels in spermatogonia and at almost undetectable levels in Sertoli cells. This expression pattern was confirmed by Northern blot analyses. Further experiments carried out on GC-1spg germ cell line revealed that reducing c-FLIPL expression increases Fas-dependent apoptosis. Conversely, restoring c-FLIPL expression reduces this response to control levels. Caspase-10 expression was found to match c-FLIPL expression pattern; further, caspase-10 activation upon anti-Fas treatment inversely correlated with c- FLIPL expression. Finally, TUNEL staining of seminiferous tubules incubated with anti-Fas antibody showed that apoptosis occurs mostly in basally located germ cells, indicating that such cells, expressing low levels of c-FLIPL, are sensitive to Fas-mediated apoptosis. These data indicate for the first time that c-FLIPL might control germ cell apoptosis and caspase activity in the adult testis.

FLIP is expressed in mouse testis and protects germ cells from apoptosis

DE CESARIS, Paola
2003

Abstract

Apoptosis control in adult testis is crucial to achieve normal spermatogenesis. In this study c-FLIP, an apoptosis-modulating protein, was investigated. In Western blot and immunohistochemical analyses, the 55 KDa c-FLIP long isoform (c- FLIPL) was found to be expressed strongly in spermatocytes and spermatids, at low levels in spermatogonia and at almost undetectable levels in Sertoli cells. This expression pattern was confirmed by Northern blot analyses. Further experiments carried out on GC-1spg germ cell line revealed that reducing c-FLIPL expression increases Fas-dependent apoptosis. Conversely, restoring c-FLIPL expression reduces this response to control levels. Caspase-10 expression was found to match c-FLIPL expression pattern; further, caspase-10 activation upon anti-Fas treatment inversely correlated with c- FLIPL expression. Finally, TUNEL staining of seminiferous tubules incubated with anti-Fas antibody showed that apoptosis occurs mostly in basally located germ cells, indicating that such cells, expressing low levels of c-FLIPL, are sensitive to Fas-mediated apoptosis. These data indicate for the first time that c-FLIPL might control germ cell apoptosis and caspase activity in the adult testis.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11697/7108
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