Localization of Tumor Necrosis Factor (TNF) in the Rat and Bovine Ovary Using Immunocytochemistry and Cell Blot: Evidence for Granulosal Production
A protein fraction recently isolated from the bovine ovary has in vitro activity similar to tumor necrosis factor-α (TNF) and was thus hypothesized to be a tumor necrosis-like factor (TNLF) (1). In another related study, rabbit corpora lutea (CL) incubated in vitro in the presence of lipopolysaccharide (LPS) secrete a substance with bioactivity similar to TNF (2). In addition, macrophages which produce TNF (3) are present within the ovary. To investigate the cellular sources of TNF in the ovary, TNF was localized in bovine and rat ovaries using polyclonal antibodies to human recombinant TNFα with a Biotin-StreptAvidin immunostaining technique. Granulosal cells were examined by cell blot technique to ascertain the presence and/or secretion of TNF using a similar Biotin-StreptAvidin immunostaining technique. This study reveals the presence of immunoreactive TNF (I-TNF) in the bovine and rat ovary localized primarily to three regions: the CL, the antral layer of granulosal cells in small antral and large preovulatory follicles, and throughout the granulosal layer of atretic follicles. Cell blot analyses indicate the presence and secretion of TNF by bovine and rat granulosa cells in vitro.
KeywordsGranulosal Cell Follicular Fluid Antral Follicle Preantral Follicle Corpus Luteum
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