Molecular mechanisms of the induction of spermatogenic cell apoptosis through mitochondria

2005 Fiscal Year Final Research Report Summary

• Project/Area Number: 15390058
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General anatomy (including Histology/Embryology)
• Research Institution: Nagasaki University
• Principal Investigator: KOJI Takehiko Nagasaki University, Graduate School of Biomedical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (30170179)
• Co-Investigator(Kenkyū-buntansha): HISHIKAWA Yoshitaka Nagasaki University, Graduate School of Biomedical Sciences, Associate Professor, 大学院・医歯薬学総合研究科, 助教授 (60304276) ; EJIMA Kuniaki Nagasaki University, Graduate School of Biomedical Sciences, Visiting Scientist, 大学院・医歯薬学総合研究科, 客員研究員 (30309984) ; IZUMI Shinichi Nagasaki University, Graduate School of Biomedical Sciences, Instructor, 大学院・医歯薬学総合研究科, 助手 (40264246)
• Project Period (FY): 2003 – 2005
• Keywords: spermatogenic cells / apoptosis / mitochondria / redistribution of Bax / cytochrome C / endocrine disruptor / electroporation / primary culture

Research Abstract

In mammalian spermatogenesis, germ cell apoptosis is very common. The mechanism to be rescued from the apoptosis seems to be essential to understand the reset of life program. Since redistribution of mitochondria derived apoptosis-related proteins, Bax and cytochrome C (cytC), in the cytoplasm was implicated in the induction of germ cell apoptosis, in the present study we investigated the distribution of mitochondria in relation with the redistribution of Bax and cytC in apoptotic germ cells in adult male mice. During the study, we also attempted to establish the method of gene transfection by electroporation in to testis in vivo and the conditions of primary culture of testicular germ cells. When the distribution of mitochondria and their proteins in apoptotic germ cells was examined by immuno-electron microscopy, the redistribution of Bax and cytC was not accompanied with a change in mitochondrial distribution. In fact, in the testis transfected with pEYFP-Mito DNA by electroporation, the fluorescence signal of YFP was co-localized with Bax and cytC, but not with SP-22 protein, which is another marker of mitochondria. These results indicate that the redistribution of apoptosis-related proteins derived from mitochondria may be triggered by loss of their ability to anchor those mitochondrial proteins. In addition, we examined various culture conditions for a long term primary culture of isolated germ cells from adult mouse testes and now can maintain the cells for several weeks in vitro, but the overall characteristics were variable among batches depending upon the quality of Sertoli cell feeder layer.

Research Products

• [Journal Article] 17-beta-estradiol protects against oxidative stress-induced cell death through the glutathione/glutaredoxin-dependent redox regulation of Akt in myocadiac H9_c2 cells. (2006)
  • Author(s): Urata, Y.
  • Journal Title: J. Biol. Chem. in press.
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 17-beta-estradiol protects against oxidative stress-induced cell death through the glutathione/glutaredoxin-dependent redox regulation of Akt in myocadiac H9c2 cells. (2006)
  • Author(s): Urata, Y., Ihara, Y., Murata, H., Goto, S., Koji, T., Yodoi, J., Inoue, S., Kondo, T.
  • Journal Title: J.Biol.Chem., (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Early human preantral follicles have relaxin and relaxin receptor (LGR7) and relaxin promotes their development. (2005)
  • Author(s): Shirota, K.
  • Journal Title: J. Clin. Endocrinol. Merab. 90・1 Pages: 516-521
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Apoptosis and its pathway in early post-implantation embryos of diabetic rats. (2005)
  • Author(s): Sun, F.
  • Journal Title: Diabetes Res. Clin. Pract. 67・2 Pages: 110-118
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Induction of epithelial cell apoptosis i the uterus a mouse uterine ischemia-reperfusion model : Possible involvement of tumor necrosis factorα. (2005)
  • Author(s): Okazaki. M.
  • Journal Title: Biol. Reprod. 72・5 Pages: 1282-1288
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Adenoviral vector-mediated gene transfer of IL-13Ralpha2 chain followed by IL-13 cytotoxin treatment offers potent targeted therapy for cytotoxin-resistant cancers. (2005)
  • Author(s): Saito, M.
  • Journal Title: Int. J Cancer 116・1 Pages: 1-8
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Induction of cell death rat small intestine by ischemia reperfusion : Differential roles of Fas/fas ligand and Bcl-2/Bax systems depending upon cell types. (2005)
  • Author(s): An, S.
  • Journal Title: Histochem. Cell Biol. 123・3 Pages: 249-261
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Early human preantral follicles have relaxin and relaxin receptor (LGR7) and relaxin promotes their development. (2005)
  • Author(s): Shirota, K., Tateishi, K., Koji, T., Hishikawa, Y., Hachisuga, T., Kuroki, M., Kawarabayashi, T.
  • Journal Title: J.Clin.Endocrinol.Metab., 90(1) Pages: 516-521
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Apoptosis and its pathway in early post-implantation embryos of diabetic rats. (2005)
  • Author(s): Sun, F., Kawasaki, E., Akazawa, S., Hishikawa, Y., Sugahara, K., Kamihira, S., Koji, T., Eguchi, K.
  • Journal Title: Diabetes Res.Clin.Pract., 67(2) Pages: 110-118
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Induction of epithelial cell apoptosis in the uterus by a mouse uterine ischemia-reperfusion model : Possible involvement of tumor necrosis factor α. (2005)
  • Author(s): Okazaki, M., Matsuyama, T., Kohno, T., Shindo, H., Koji, T., Morimoto, Y., Ishimaru, T
  • Journal Title: Biol.Reprod., 72(5) Pages: 1282-1288
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Adenoviral-mediated gene transfer of IL-13R α2 chain followed by IL-13 cytotoxin treatment offers potent targeted therapy for cytotoxin-resistant cancers. (2005)
  • Author(s): Saito, M., Murata, T., Watanabe, K., Kawakami, K., Suzuki, M., Koji, T., Puri, R.K., Kitazato, K., Kobayashi, N.
  • Journal Title: Int.J.Cancer, 116(1) Pages: 1-8
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Induction of cell death in rat small intestine by ischemia-reperfusion : differential roles of Fas/Fas ligand and Bcl-2/Bax systems depending upon cell types. (2005)
  • Author(s): An, S., Hishikawa, Y., Koji, T.
  • Journal Title: Histchem.Cell Biol., 123(3) Pages: 249-261
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The cognition-enhancer nefiracetam inhibits both necrosis and apoptosis in retinal ischemic models in vitro and in vivo. (2004)
  • Author(s): Ueda, M.
  • Journal Title: J. Pharmacol. Exp. Ther. 309・1 Pages: 200-207
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification and localization of estrogen receptor α-and β-positive cells in adult male and female mouse intestine at various estrogen levels. (2004)
  • Author(s): Kawano, N
  • Journal Title: Histochem. cell biol. 121・5 Pages: 399-405
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Expression of keratinocyte growth factor/fibroblast growth factor-7 and its receptor in human lung cancer. correlation with tumour proliferative activity and patient prognosis. (2004)
  • Author(s): Yamayoshi, T.
  • Journal Title: J. Pathol. 204・1 Pages: 110-118
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Estrogen receptor-associated expression of keratinocyte growth factor and its possible role in the inhibition of apoptosis in human breast cancer. (2004)
  • Author(s): Tamaru, N.
  • Journal Title: Lab. Invest. 84・11 Pages: 1460-1471
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The cognition-enhancer nefiracetam inhibits both necrosis and apoptosis in retinal ischemic models in vitro and in vivo. (2004)
  • Author(s): Ueda, M., Fujita, R., Koji, T., Ueda, H.
  • Journal Title: J.Pharmacol.Exp.Ther., 309(1) Pages: 200-207
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification and localization of estrogen receptor α- and β-positive cells in adult male and female mouse intestine at various estrogen levels. (2004)
  • Author(s): Kawano, N., Koji, T., Hishikawa, Y., Murase, K., Murata, I., Kohno, S.
  • Journal Title: Histochem.Cell Biol., 121 Pages: 399-405
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Expression of keratinocyte growth factor/fibroblast growth factor-7 and its receptor in human lung cancer : correlation with tumour proliferative activity and patient prognosis. (2004)
  • Author(s): Yamayoshi, T., Nagayasu, T., Matsumoto, K., Abo, T., Hishikawa, Y., Koji, T.
  • Journal Title: J.Pathol., 2004(1) Pages: 110-118
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Estrogen receptor-associated expression of keratinocyte growth factor and its possible role in the inhibition of apoptosis in human breast cancer. (2004)
  • Author(s): Tamaru, N., Hishikawa, Y., Ejima, K., Nagasaue, N., Inoue, S., Muramatsu, M., Hayashi, T., Koji, T.
  • Journal Title: Lab.Invest., 84(11) Pages: 1460-1471
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Grem cell apoptosis and its molecular trigger in mouse testes (Review) (2003)
  • Author(s): Koji, T.
  • Journal Title: Arch. Histol. Cytol. 66・1 Pages: 1-16
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Molecular histochemical analysos of estrogen receptor α and β expressions in the mouse ovary : In situ hybridization and Southwestern histochemistry. (2003)
  • Author(s): Hishikawa, Y.
  • Journal Title: Med. Electron Microsc. 36・2 Pages: 67-73
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Possible involvement of keratinocyte growth factor and its receptor in enhanced epithelial-cell proliferation and acquired recurrence if middle-ear cholesteatoma. (2003)
  • Author(s): Yamamoto-Fukuda, T.
  • Journal Title: Lab. Invest. 83・1 Pages: 123-136
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Zone-dependent expression of estrogen receptor α and β in human benign prostatic hyperplasia. (2003)
  • Author(s): Tsurusaki, T.
  • Journal Title: J. Clin. Endocrinol. Metab. 88・3 Pages: 1333-1340
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Antisense oligonucleotides against collagen-binding stress protein HSP47 suppress peritoneal fibrosis in rats. (2003)
  • Author(s): Nishino, T.
  • Journal Title: Kidney Int. 64・3 Pages: 887-896
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Germ cell apoptosis and its molecular trigger in mouse testes (Review). (2003)
  • Author(s): Koji.T., Hishikawa, Y.
  • Journal Title: Arch.Histol.Cytol., 66(1)(and Cover) Pages: 1-16
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Molecular histochemical analysis of estrogen receptor α and β expressions in the mouse ovary : In situ hybridization and Southwestern histochemistry. (2003)
  • Author(s): Hishikawa, Y., Damavandi, E., Izumi, S., Koji T.
  • Journal Title: Med.Electron Microsc., 36(and Cover) Pages: 67-73
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Possible involvement of keratinocyte growth factor and its receptor in enhanced epithelial-cell proliferation and acquired recurrence of middle-ear cholesteatoma. (2003)
  • Author(s): Yamamoto-Fukuda, T., Aoki, D., Hishikawa, Y., Kobayashi, T., Takahashi, H., Koji, T.
  • Journal Title: Lab.Invest., 83(1) Pages: 123-136
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Zone-dependent expression of estrogen receptors α and β in human benign prostatic hyperplasia. (2003)
  • Author(s): Tsurusaki, T., Aoki, D., Kanetake, H., Inoue, S., Muramatsu, M., Hishikawa, Y., Koji, T.
  • Journal Title: J.Clin.Endocrinol.Metab., 88(3) Pages: 1333-1340
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Antisense oligonucleotides against collagen-binding stress protein HSP47 suppress peritoneal fibrosis in rats. (2003)
  • Author(s): Nishino, T., Miyazaki, M., Abe, K., Furusu, A., Mishima, Y., Harada, T., Ozono, Y., Koji.T., Kohno, S.
  • Journal Title: Kidney Int., 64(3) Pages: 887-896
  • Description: 「研究成果報告書概要(欧文)」より