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and Its Receptor in Bovine Corpus Luteum Throughout the Estrous Cycle1
a Laboratory of Reproductive Endocrinology, Faculty of Agriculture, Okayama University, Okayama 700-8530, Japan
b Institute of Physiology, Technical University of Munich, D-85350 Freising-Weihenstephan, Germany
c Laboratory of Animal Reproduction, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan
The objective of this study was to investigate tumor necrosis factor 
(TNF-) expression, the presence of functional TNF- receptors, and expression of TNF receptor type I (TNF-RI) mRNA in the bovine corpus luteum (CL) during different stages of the estrous cycle. Reverse transcription (RT)-polymerase chain reaction (PCR) showed no difference in TNF- mRNA expression during the estrous cycle. Concentrations of TNF- in the CL tissue increased significantly from the mid to the late luteal stage and decreased thereafter (P < 0.05). An RT-PCR analysis showed higher levels of TNF-RI mRNA in CL of Days 3–7 than of other stages (P < 0.05). 125I-TNF- binding to the membranes of bovine CL was maximal after incubation at 38°C for 48 h. The binding was much greater for TNF- than for related peptides. A Scatchard analysis revealed the presence of a high-affinity binding site in the CL membranes collected at each phase of the estrous cycle (dissociation constant: 3.60 ± 0.58–5.79 ± 0.19 nM). In contrast to TNF-RI mRNA expression, the levels of receptor protein were similar at each stage of the estrous cycle. When cultured cells of all luteal stages were exposed to TNF- (1–100 ng/ml), TNF- stimulated prostaglandin F2 and prostaglandin E2 secretion by the cells in a dose-dependent fashion (P < 0.01), especially during the early luteal phase, although it did not affect progesterone secretion. These results indicate the local production of TNF- and the presence of functional TNF-RI in bovine CL throughout the estrous cycle, and suggest that TNF- plays some roles in regulating bovine CL function throughout the estrous cycle.
1 This research was supported by Grants-in-Aid for Scientific Research (No. 11460129) and a Research Fellowship (No. 8763) of the Japan Society for the Promotion of Science (JSPS) from the Ministry of Education, Science, Sports and Culture of Japan; the Japanese-German Cooperative Science Promotion Program of JSPS; and the German Research Foundation (Scha 257/14–1).
2 Correspondence: Kiyoshi Okuda, Laboratory of Reproductive Endocrinology, Division of Animal Science and Technology, Faculty of Agriculture, Okayama University, Okayama 700-8530, Japan. FAX: 81 86 251 8388; kokuda{at}cc.okayama-u.ac.jp
3 Current address: Department of Animal Reproduction, College of Agriculture, Osaka Prefecture University, Sakai 599-8531, Japan.
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