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Regular Article |
a Laboratory of Reproductive Endocrinology, Department of Animal Science, Faculty of Agriculture, Okayama University, Okayama 700-8530, Japan
b Department of Immunology, National Institute of Animal Health, Ibaraki 305-0856, Japan
Fas antigen (Fas) is a cell surface receptor that triggers apoptosis in sensitive cells when bound to the Fas ligand (Fas L). The present study was undertaken to identify the presence of a Fas-Fas L system in bovine corpus luteum (CL) and to evaluate the regulation of Fas-mediated luteal cell death by leukocyte-derived cytokines. The reverse transcription-polymerase chain reaction showed higher levels of Fas mRNA expression in CL in the regressed luteal stage (Days 19–21) than in the other stages (P < 0.05). Bovine luteal cells from midcycle CL (Days 8–12) were exposed for 24 h to interferon 
(IFN; 50 ng/ml) and/or tumor necrosis factor 
(TNF; 50 ng/ml). After 24 h of culture, the expression of Fas mRNA was detected in the cultured cells and was increased by IFN. Moreover, TNF augmented the stimulatory action of IFN, whereas TNF alone did not affect the expression of Fas mRNA. The effects of IFN and TNF on Fas-mediated cell death were also examined. Cells were exposed to IFN and/or TNF for 24 h and were further treated with IFN and/or TNF in the presence or absence of Fas L (100 ng/ml) for 24 h. Treatments of the cells with IFN alone and in combination with TNF resulted in killing of 30% and 50% of the cells (P < 0.05), respectively, whereas TNF alone did not have a cytotoxic effect on the cells. On the other hand, Fas L killed 60% of the cells treated with IFN (P < 0.01) and 85% of the cells treated with the combination of TNF and IFN (P < 0.01), respectively, whereas Fas L showed no effect on the viability of the luteal cells treated with or without TNF. Furthermore, shrunken nuclei and apoptotic bodies were observed in the cells treated with Fas L in the presence of TNF and IFN. The overall results suggest that a Fas-Fas L system is present in bovine CL and that leukocyte-derived TNF and IFN play important roles in Fas-mediated luteal cell death.
1 This research was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan (No. 11460129) and Grants-in-Aid for the Recombinant Cytokine Project provided by Ministry of Agriculture, Forestry and Fisheries of Japan.
2 Correspondence. FAX: 81 86 251 8388; kokuda{at}cc.okayama-u.ac.jp
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