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b Departments of Theriogenology
c and Animal Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080, Japan
Each preovulatory follicle (selected based on the concentrations of steroids and PGE2) was dissected from surrounding stromal tissue and implanted with 4 capillary dialysis membranes (control, LH, cytokines or ET-1, and LH+cytokine or LH+ET-1) into the theca layer. They were then incubated in organ culture chambers and perfused with Ringer's solution for 14 h after pre-perfusion for 2 h. The stimulation with LH (5 µg/ml) between 4 and 6 h increased the release of progesterone (P4), androstenedione (A), estradiol-17ß (E2), PGE2 (p < 0.001), and ET-1 (p < 0.05). The infusion of ET-1 (250 ng/ml) between 8 and 10 h inhibited P4 and A release but stimulated E2 release (p < 0.05). The infusion of TNF
These results show that ET-1 is released from the theca layer of mature bovine follicles in vitro and that it affects follicular steroids and PGE2 secretion. The overall results suggest that interactions among ET-1, PGE2, and cytokines may have key roles in a local intermediatory/amplifying system of the LH-triggered ovulatory cascade in the bovine preovulatory follicle.
Local regulation of ovulation involves the interaction of LH and intrafollicular factors including steroids, prostaglandins, and peptides derived from endothelial cells, leukocytes, fibroblasts, and steroidogenic cells. To estimate the intrafollicular role of endothelin-1 (ET-1) and its possible interaction with LH, tumor necrosis factor 
(TNF), and interleukin-1ß (IL-1ß), a microdialysis system was implanted into the theca layer of preovulatory bovine follicles that were maintained in organ culture chambers. The effects of LH, ET-1, TNF, and IL-1ß on the local release of steroids, prostaglandin E2 (PGE2), and ET-1 from the cells surrounding the implanted capillary membrane were investigated. (100 ng/ml) between 8 and 10 h after LH exposure suppressed the release of A and E2 (p < 0.05). IL-1ß (10 ng/ml) between 8 and 10 h stimulated E2 release but inhibited A release (p < 0.05). Moreover, ET-1 and cytokines clearly stimulated PGE2 release (p < 0.05). ET-1 and TNF induced further release of PGE2 stimulated by LH (p < 0.05). Also, TNF and IL-1ß induced further release of ET-1 stimulated by LH (p < 0.05).
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