Tumor necrosis factor alpha inhibits ovulation and induces granulosa cell death in rat ovaries
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ORIGINAL ARTICLE
Tumor necrosis factor alpha inhibits ovulation and induces granulosa cell death in rat ovaries Yuri Yamamoto • Akira Kuwahara • Yuka Taniguchi • Mikio Yamasaki Yu Tanaka • Yukari Mukai • Mizuho Yamashita • Toshiya Matsuzaki • Toshiyuki Yasui • Minoru Irahara
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Received: 27 June 2014 / Accepted: 26 November 2014 Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Purpose We evaluated the role of tumor necrosis factor alpha (TNFa) in rat ovulation and granulosa cell death of ovarian follicles during the periovulatory stage. Methods Immature rats primed with pregnant mare serum gonadotropin were injected intraperitoneally with human chorionic gonadotropin (hCG), and TNFa was injected into the bursa 48 h later. The total number of released oocytes was counted. Apoptosis was measured with terminal
Y. Yamamoto A. Kuwahara (&) Y. Taniguchi M. Yamasaki Y. Tanaka Y. Mukai M. Yamashita T. Matsuzaki T. Yasui M. Irahara Department of Obstetrics and Gynecology, The University of Tokushima, Institute for Health Biosciences, 3-18-15 Kuramoto-cho, Tokushima 770-8503, Japan e-mail: [email protected] Y. Yamamoto e-mail: [email protected] Y. Taniguchi e-mail: [email protected]
deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and the expression of cleaved caspase 3 and Bax/Bcl-2. Autophagy was assessed by the expression of light chain protein 3 (LC3) and autophagosomes under transmission electron microscopy. Results TNFa significantly decreased the number of released oocytes, and many unruptured follicles were observed. TUNEL analysis revealed a larger number of apoptotic cells, and the cleaved caspase 3 and Bax/Bcl-2 increased more than that of the control 12 h after hCG administration. Furthermore, the expression of LC3 wwas significantly higher than that of the control, and autophagosomes were observed in the cytoplasm. Conclusions Our data indicated that TNFa is an important mediator of ovulation in terms of decreasing the number of released oocytes and inducing granulosa cell death of unruptured follicles via apoptosis and autophagy for remodeling ovarian tissues. Keywords Apoptosis Autophagy Granulosa cell death Ovulation TNF-alpha
M. Yamasaki e-mail: [email protected] Y. Tanaka e-mail: [email protected]
Introduction
Y. Mukai e-mail: [email protected]
Tumor necrosis factor alpha (TNFa), a nonglycosylated protein with a molecular weight of 17 kDa, is produced by activated macrophages [1]. TNFa is a multifunctional cytokine and mediates a wide range of biological actions that include not only the regulation of proinflammatory responses but also the control of cell differentiation, tissue renewal, and restructuring [2]. Recent studies have demonstrated that TNFa plays a role in ovarian follicular development [3], steroidogenesis [4, 5], ovulation [5, 6], luteolysis [7], and atresia [8, 9]. TNFa is localized in human oocytes and
M. Yamashita e-mail: mizu-ton@m
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