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This study aimed to investigate the presence of polymorphonuclear neutrophils (PMNs) in bovine oviduct fluid under physiological conditions and to determine the possible role of bovine oviduct epithelial cells (BOECs) in the regulation of the phagocytic activity of PMNs for sperm. During the pre-ovulatory stage, PMNs were identified in the bovine oviduct fluid in relatively constant numbers. In our experiments, PMNs were incubated for 4 h with the supernatant of cultured BOECs stimulated for 24 h by LH (10 ng/ml). Phagocytosis was then assayed by co-incubation of these PMNs with sperm treated to induce capacitation. The BOEC supernatant significantly suppressed sperm phagocytosis by PMNs, and the LH-stimulated BOEC supernatant further suppressed phagocytosis. Importantly, in the BOEC culture, LH stimulated the secretion of prostaglandin E 2 (PGE 2 ), which dose-dependently (10 K6 , 10 K7 , and 10 K8 M) suppressed sperm phagocytosis by PMNs. Furthermore, a PGEP 2 receptor antagonist significantly abrogated the inhibition of phagocytosis by the LH-stimulated BOEC supernatant. Additionally, using scanning electron microscopy, incubation of PMNs with either PGE 2 or LH-stimulated BOEC supernatant before phagocytosis was found to prevent the formation of DNA-based neutrophil extracellular traps for sperm entanglement. The results indicate that sperm are exposed to PMNs in the oviduct and PGE 2 released into the oviduct fluid after LH stimulation may play a major role in the suppression of the phagocytic activity of PMNs for sperm via interaction with EP 2 receptors. Thus, the bovine oviduct provides a PGE 2 -rich microenvironment to protect sperm from phagocytosis by PMNs, thereby supporting sperm survival in the oviduct.Free Japanese abstract A Japanese translation of this abstract is freely available at

The precise regulatory mechanisms of cyclic oviductal contraction in the cow are unclear. The purpose of this study was to evaluate the effect of luteinizing hormone (LH), steroids, prostaglandins (PGs) and peptides on the oviductal contraction and secretion of PGs and endothelin (ET-1). In addition, the cyclic expression of mRNA for ET-1 and its receptors (ET-R) was evaluated by reverse transcription-polymerase chain reaction (RT-PCR). In the in vitro microdialysis study, an infusion of LH alone or in combination with progesterone (P 4 ), estradiol-17 (E 2 ) and/or ET-1 stimulated pronounced release of PGE 2 , PGF 2 and ET-1 in the oviducts from cows in the follicular and postovulatory phases. The addition of LH, LH+P 4 +E 2 and/or ET-1 to the medium increased the amplitude of oviductal contraction. However, oxytocin (OT) completely blocked the responses of oviductal secretion and contraction. In contrast, these substances did not show any effect in the oviducts from cows in the mid luteal phase. Similar expression patterns of mRNA encoding for ET-R type A and type B were found, which were highest during the postovulatory phase, lower during the luteal phase, with the lowest expression during the follicular phase. We suggest that the preovulatory LH surge, together with increasing E 2 levels from the Graafian follicle and a basal P 4 from regressing corpora lutea (CL), stimulates maximum oviductal production of PG and ET-1, resulting in oviductal contraction for a rapid transport of gametes. OT released from the newly-formed CL may block these mechanisms, and slow contractions for transport of the embryo to the uterus.

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 alpha (TNFalpha), and interleukin-1beta (IL-1beta), 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, TNFalpha, and IL-1beta on the local release of steroids, prostaglandin E2 (PGE2), and ET-1 from the cells surrounding the implanted capillary membrane were investigated. 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 microg/ml) between 4 and 6 h increased the release of progesterone (P4), androstenedione (A), estradiol-17beta (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 TNFalpha (100 ng/ml) between 8 and 10 h after LH exposure suppressed the release of A and E2 (p < 0.05). IL-1beta (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 TNFalpha induced further release of PGE2 stimulated by LH (p < 0. 05). Also, TNFalpha and IL-1beta induced further release of ET-1 stimulated by LH (p < 0.05). 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.