Currently, histological classifications of ovarian follicular atresia are almost exclusively based on the morphology of the membrana granulosa without reference to the theca interna. Atresia in the bovine small antral ovarian follicle has been redefined into antral or basal atresia where cell death commences initially within antral or basal regions of the membrana granulosa, respectively. To examine cell death in the theca interna in the two types of atretic follicles, bovine ovaries were collected and processed for immunohistochemistry and light microscopy. Follicles were classified as healthy, antral atretic, or basal atretic. Follicle diameter was recorded and sections stained with lectin from Bandeiraea simplicifolia to identify endothelial cells or with an antibody to cytochrome P450 cholesterol side-chain cleavage to identify steroidogenic cells and combined with TUNEL labeling to identify dead cells. The numerical density of steroidogenic cells within the theca interna was significantly reduced (P < 0.001) in basal atretic follicles in comparison with other follicles. Cell death was greater in both endothelial cells (P < 0.05) and steroidogenic cells (P < 0.01) of the theca interna of basal atretic follicles compared with healthy and antral atretic follicles. Thus, we conclude that the theca interna is susceptible to cell death early in atresia, particularly in basal atretic follicles.

SUMMARY Gardnerella vaginalis was ingested and killed by neutrophils in the presence of normal human serum. Heat inactivation of the serum inhibited these processes. The opsonisation of some but not all G vaginalis strains was enhanced by immune rabbit serum. Immune serum did not, however, enhance intracellular killing. Blockade of the classical pathway of complement activation had no effect on the opsonic activity of human serum. These results suggest that the opsonisation, phagocytosis, and killing of G vaginalis by human serum and neutrophils is primarily mediated by complement activated by the alternative pathway. Serum without neutrophils has little effect on the viability of G vaginalis. Human neutrophils were separated from venous blood by dextran sedimentation and residual red cells were lysed by treatment wth 0-83% Tris buffered ammonium chloride.9 After a wash, neutrophils were counted and resuspended to a concentration of 106 cells/ml in HBSS without phenol red.For microscopical assays bacteria and cells were incubated at a ratio of 5:1 for 15 min at 37°C. Smears were then prepared using a cytospin centrifuge (Shandon) and stained with Giemsa.

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