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Immunohistochemical localization and possible physiological roles of tumor necrosis factor-alpha in mouse cumulus-oocyte complexes

, : Immunohistochemical localization and possible physiological roles of tumor necrosis factor-alpha in mouse cumulus-oocyte complexes. Development Growth & Differentiation 37(4): 413-420

Tumor necrosis factor-alpha (TNF-alpha), a cytokine which is produced by activated macrophages, has been shown to participate in the regulation of ovarian functions. In the course of our investigation on the mechanism of maturation, fertilization and degeneration of mouse oocytes, immunoreactivity to TNF-alpha was found in the cytoplasm of the cells surrounding the maturing oocytes and of granulosa cells facing the antral cavity. Immunoblot analysis with the specific antibody to TNF-alpha identified the 17 kDa Mr band in the extract of cumulus-oocyte complexes. Various concentrations of TNF-alpha (mouse, recombinant) and anti TNF-alpha antiserum (polyclonal rabbit anti-mouse recombinant TNF-alpha) were then used to determine their effect on the germinal vesicle breakdown (GVBD), polar body extrusion, fertilization and fragmentation of mouse oocytes/eggs. TNF-alpha at concentrations of 10 ng/mL or less and anti-TNF-alpha antiserum at concentrations of 10% or less, had no effect on the spontaneous GVBD and polar body extrusion of mouse oocytes in culture. Mouse follicular oocytes cultured for more than 72 h in modified Krebs-Ringer solution in vitro undergo spontaneous fragmentation, which is a degenerative change to form 'blastomeres' with or without nuclear fragments or chromatin. Ghost-like blastomeres were also identified in the space among fragmented 'blastomeres'. The spontaneous fragmentation of mouse follicular oocytes was suppressed in the presence of TNF-alpha at concentrations of 1 ng/mL or greater. Anti-TNF-alpha antiserum (1%) accelerated the induction of fragmentation of oocytes cultured in vitro. The addition of anti TNF-alpha antiserum (10%) to the culture medium did not influence the fertilization rates of the eggs surrounded by the expanded cumulus. These results appear to indicate that the process of degeneration of mouse oocytes/eggs is modulated by TNF-alpha accumulated in the expanded cumulus during oocyte maturation.


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