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Plasmin Cleaves Tumor Necrosis Factor Exodomain from Sheep Follicular Endothelium: Implication in the Ovulatory Process¹

^a Department of Animal Science, University of Wyoming, Laramie, Wyoming 82071

Ovulation in the sheep is predicated on plasmin up-regulation at the ovarian surface-follicular interface, release of tumor necrosis factor (TNF) from contiguous endothelium, and apoptotic cell death. The objectives of this investigation were to determine whether plasmin elicits TNF secretion from thecal endothelium of ovine follicles, to characterize the site(s) of enzymatic attack, and to assess the physiological consequence of soluble TNF action. Endothelial cells of thecal tissues isolated from antral follicles of eCG-primed anestrous ewes shed (histochemical depletion) TNF into incubation medium (ovarian cell DNA fragmentation bioassay, Western blot detection) upon exposure to plasmin. Immunopurification and N-terminal sequence analysis indicated that TNF was excised from its transmembrane precursor at the Arg79-Ser80 and Lys88-Pro89 linkages. Microinjection of TNF into the apical wall of explanted follicles induced cellular apoptosis and stigma development. We suggest that plasmin-mediated cleavage of TNF exodomain from its membrane anchor along thecal endothelium is a determinant of tissue dissolution within the formative ovulatory rupture site of ewes.

¹ Supported by USDA-NRI grant 95–37203–2131.

² Correspondence: W.J. Murdoch, Department of Animal Science, P.O. Box 3684, 16th and Gibbon St., University of Wyoming, Laramie, WY 82071. FAX: 307 766 2355; wmurdoch{at}uwyo.edu

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