Detection of the activation of caspases during neuronal cell death by using antiserum specifically recognizing active form of caspases

• Project/Area Number: 11480235
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: National Institute of Neuroscience
• Principal Investigator: MOMOI Takashi National Institute of Neuroscience Div. Development and Differentiation, Lab. Head, 神経研究所・疾病研究第5部, 室長 (40143507)
• Co-Investigator(Kenkyū-buntansha): 内山 安男 大阪大学, 大学院医学系・研究科・情報伝達医学機能形態学, 教授 (10049091) ; 塚原 俊文 国立精神・神経センター, 神経研究所・疾病研究第1部, 研究員 (60207339)
• Project Period (FY): 1999 – 2002
• Project Status: Completed (Fiscal Year 2002)
• Budget Amount: ¥11,800,000 (Direct Cost: ¥11,800,000); Fiscal Year 2002: ¥2,100,000 (Direct Cost: ¥2,100,000); Fiscal Year 2001: ¥2,800,000 (Direct Cost: ¥2,800,000); Fiscal Year 2000: ¥2,800,000 (Direct Cost: ¥2,800,000); Fiscal Year 1999: ¥4,100,000 (Direct Cost: ¥4,100,000)
• Keywords: apoptosis / caspase-12 / ER stress / polyglutamine disease / Huntinton / Endoplasmic Reticulum / processing / Neurodegenerative disease / カスパーゼー12 / ER ストレス / ハンチントン病 / ポリグルタミン / カスパーゼ12 / 神経細胞死 / 脳形成 / カスパーゼ9 / カスパーゼ3 / Bcl-x / ミトコンドリア / チトクロームC / Akt / BMP-4 / レチノイン酸

Research Abstract

Considerable evidence suggests that caspase, a member of cystein protease, plays an essential role in the various cell death including apoptosis and programmed cell death (PCD) and in the cell death observed in the various diseases. However, there is little direct evidence on the involvement of caspases in naturally occurring cell death during development of the vertebrate nervous system and diseases. Caspases are sequentially activated in the caspase cascade, in which activated initial caspases such as caspase 9, caspase 8 and caspase 12 are autoprocessed and then process caspase 3, 6 and 7. To clarify the involvement of caspases in the PCD and neurodegenerative diseases, we prepared cleavage site-directed antiserum against 12 members of caspases that specifically react with the cleaved fragment of caspases, respectively. We have examined directly the activation of caspases during development of mouse embryos and subcellular localization of active form of caspases by immunohistochemical staining using these antisera.

Research Products

• [Publications] Momoi, T., Fujita, E., Urase, K.: "Strain-specific caspase-3-dependent programmed cell death in the developing forebrain"Neuroreport. 14. 111-115 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Jimbo, A., Fujita, E., Kouroku, Y., Ohnishi, J., Inohara, N., Kuida, K., Sakamaki, K., Yonehara, S., Momoi, T.: "ER stress induces caspase-8 activation stimulating the cytochrome c release and caspase-9 activation"Exp.Cell.Res.. 283. 156-166 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Fujita, E., Kouroku, Y., Jimbo, A., Maruyama, K., Momoi, T.: "Caspase-12 processing and fragment translocation into nuclei of tunicamycin-treated cells"Cell Death Differ.. 9. 1108-1114 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kouroku, Y., Fujita, E., Jimbo, A., Kikuchi, T., Yamagata, T., Momoi, MY, Kominami E., Kuida, K., Sakamaki, K., Yonehara, S., Momoi, T.: "Polyglutamine aggregates stimulate ER stress signals and caspase-12 activation"Hum.Mol.Genet.. 11. 1505-1515 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Fujita, E., Egashira, J., Urase, K., Kuida, K., Momoi, T.: "Caspase-9 processing via feedback amplification loop in vivo"Cell Deth and Differentiation. 8. 335-344 (2001)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Takahashi, K., Kamiya, K., Urase, K., Fujita, E., Suga, M., Mori, H., Yoshikawa, Y., Ichimura, K., Kuida, K., Momoi, T.: "Caspase-3-deficiency induces hyperplasia of supporting cells and degeneration of sensory cells resulting in the hearing loss"Brain Res.. 894. 359-367 (2001)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kouroku Y, Fujita E, Urase K, Tsuru T, Setsuie R, Kikuchi T, Yagi Y, Momoi MY, Momoi T.: "Caspase that are activated during generation of nuclera pologlutamine aggregates are necessary for DNA fragmentation but not suffiviennt for cell death"J Neurosci Res.. 62. 547-556 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Fujita E, Urase K, Egashira J, Miho Y, Isahara K, Uchiyama Y, Isoai A, Kumagai H, Kuida K, Motoyama N, Momoi T.: "Detection of caspase-9 activation in the cell death of the Bcl-x-deficient mouse embryo nervous system by cleavage sites-directed antisera"Brain Res Dev Brain Res.. 122. 135-147 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kouroku Y, Fujita E, Jimbo A, Mukasa T, Tsuru T, Momoi MY, Momoi T.: "Localization of active form of caspase-8 in mouse L929 cells induced by TNF treatment and polyglutamine aggregates"Biochem Biophys Res Commun. 270. 270,972-977 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Fujita E, Soyama A, Kuwabata M, Momoi T: "BMP-4 and retinoic acid synergistically induce activation of caspase-9 and cause apoptosis of P19 embryunal carcinoma cells cultured as a monolayer"Cell Death Differ.. 6. 1109-1116 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Momoi, T., Fujita, E., and Urase, K.: "Strain-specific caspase-3-dependent programmed cell death in the developing forebrain"Neuroreport. 14. 111-115 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Jimbo, A., Fujita, E., Kouroku, Y., Ohnishi, J., Inohara, N., Kuida, K., Sakamaki, K., Yonehara, S., and Momoi, T.: "ER stress induces caspase-8 activation stimulating the cytochrome c release and caspase-9 activation"Exp. Cell Res.. 283. 156-166 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Fujta, E., Kouroku, Y., Jimbo, A., Maruyama, K., and Momou, T.: "Caspase-12 processing and fragment translocation into nuclei of tunicamycin-treated cells"Cell Death Differ. 9. 1108-1114 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kouroku, Y., Fujita, E., Jimbo, A., Kikuchi, T., Yamagata, T., Momoi, M..Y., Kominami, E., Kuida, K., Sakamaki, K., Yonehara, S., and Momoi, T.: "Polyglutamine aggregates stimulate ER stress signals and caspase-12 activation"Human Mol. Genet. 11. 1505-1515 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Fujita, E., Egashira, J., Urase, K., Kuida, K., Momoi, T.: "Caspase-9 processing via feedback amplification loop in vivo"Cell Death and Differentiation. 8. 335-344 (2001)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Takahashi, K., Kamiya, K., Urase, K., Suga, M., Takizawa, T., Mori, H., Yoshikawa, Y., Ichimura, K., Kuida, K., Momoi, T.: "Caspase-3-deficiency induces hyperplasia of supporting ceils and degeneration of sensory cells resulting in the hearing loss"Brain Res. 894. 359-367 (2001)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kouroku Y, Fujita E, Urase K, Tsuru T, Setsuie R, Kikuchi T, Yagi Y, Momoi MY, Momoi T: "Caspases that are activated during generation of nuclear polyglutamine aggregates are necessary for DNA fragmentation but not sufficient for cell death"J Neurosci Res. 62. 547-556 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Fujita E, Urase K, Egashira J, Miho Y, Isahara K, Uchiyama Y, Isoai A, Kumagai H, Kuida K, Motoyama N, Momoi T: "Detection of caspase-9 activation in the cell death of the Bcl-x-deficient mouse embryo nervous system by cleavage sites-directed antisera"Brain Res Dev Brain Res. 122. 135-147 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kouroku Y, Fujita E, Jimbo A, Mukasa T, Tsuru T, Momoi MY, Momoi T: "Localization of active form of caspase-8 in mouse L929 cells induced by TNF treatment and polyglutamine aggregates"Biochem Biophys Res Commun. 270. 972-977 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Fujita E, Soyama A, Kawabata M, Momoi T: "BMP-4 and retinoic acid synergistically induce activation of caspase-9 and cause apoptosis of P19 embryonal carcinoma cells cultured as a monolayer"Cell Death Differ. 6. 1109-1116 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Momoi T., Fujita, E., Urase, K.: "Region-specific caspase-3-dependent programmed cell death in the developing forebrain"Neuroreport. 14. 111-115 (2003)
• [Publications] Jimbo, A., Fujila, B., Kouroku, Y., Ohnishi, J., Inohara, N., Kuida, K., Sakainaki, K., Yonehara.S., Momoi, T.: "ER stress induces caspase-8 activation stimulating the cytochrome c release and caspase-9 activation"Exp. Cell. Res.. 283. 156-166 (2003)
• [Publications] Kitanaka C, Kato K, Ijiri R, Sakurada K, Tomiyama A, Noguchi K, Nagashima Y, Nakagawara A, Momoi T, Toyoda Y, Kigasawa H, Nishi T, Shirouzu M, Yokovama S, Tanaka Y, Kuchino Y.: "Increased ras expression and caspase independent neuroblastoma cell death : possible mechanism of spontaneous neuroblastoma regression"J. Nail. Cancer Inst.. 94. 358-368 (2002)
• [Publications] Nakamoto Y, Kaneko S, Fan Ii, Mornoi T, Tsutsui H, Nakanishi K, Kobayashi K, Suda T.: "Prevention of Hepatocellular Carcinoma Development Associated with Chronic Hepatitis by Anti-Fas Ligand Antibody Therapy"Exp. Med.. 196. 1105-1111 (2002)
• [Publications] Fujita, E., Kouroku, Y., Jimbo, A., Maruyaina, K., Moinoi, T.: "Caspase-12 processing and fragment translocation into nuclei of tunicamycin-treated cells"Cell Death Differ.. 9. 1108-1114 (2002)
• [Publications] Kouroku, Y., Fujita, E., Jiznbo, A., Kikuchi, T., Yainagata, T., Mornoi, MY, Korninarni E., Kuida, K., Sakamaki, K., Yonehara.S., Mornoi.T.: "Polyglutamine aggregates stimulate ER stress signals and caspase-12 activation"Hum. Mol. Genet.. 11. 1505-1515 (2002)
• [Publications] Fujita E et al.: "Caspase-9 processing via feedback amplification loop in vivo."Cell Death and Differentiation.. 8. 335-344 (2001)
• [Publications] Inada H et al.: "Keratin attenuates tumor necrosis factor-induced cytotoxicity through association with TRADD."Cell Biol.. 155. 415-426 (2001)
• [Publications] Kitanaka C et al.: "Increased ras expression and caspase-independent neuroblastoma cell death : possible mechanism of spontaneous neuroblastoma regression."J Natl Cancer Inst.. 94. 358-368 (2002)
• [Publications] Shigemura N et al.: "Localization of activated caspase-3-positive and apoptotic cells in the developing tooth germ of the mouse lower first molar"Histochem J.. 33. 253-258 (2001)
• [Publications] Yaginuma H et al.: "Caspase activity is involved in, but is dispensable for, early motoneuron death in the chick embryo cervical spinal cord."Mol Cell Neurosci.. 18. 168-182 (2001)
• [Publications] Hisahara S et al.: "Caspase-11 mediates oligodendrocyte cell death and pathogenesis of autoimmune-mediated demyelination."J Exp Med.. 193. 111-122 (2001)
• [Publications] Fujita,E. et al.: "Caspase-9 processing via feedback amplification loop in vivo."Cell Death and Differentiation. (In press). (2001)
• [Publications] Hisahara,S. et al.: "Caspase-11 mediates oligodendrocyte cell death and pathogenesis of autoimmune-mediated demyelination."J.Exp.Med.. 193. 111-122 (2001)
• [Publications] Zheng,TS. et al.: "Deficiency of caspase-9 or caspase-3 induces the compensation of caspase."Nature Medicine. 6. 2141-2147 (2000)
• [Publications] Fujita,E. et al.: "Activation of caspase-9 in the nervous systems of Bcl-xL deficient mice embryos."Dev.Brain Res.. 122. 135-147 (2000)
• [Publications] Tanaka,M. et al.: "In situ detection of activated caspase-3 apoptotic granule neurons in the developing cerebellum in slice culture and in vivo."Dev.Brain Res.. 121. 223-228 (2000)
• [Publications] Ikezoe,K. et al.: "A novel congenital myopathy with apoptotic changes."Acta.Neurology. 47. 531-536 (2000)
• [Publications] Miho Y., et al.: "bFGF inhibits the activation of caspase-3 and apoptosis of P19 embryonal carcinoma cells during neuronal differentiation"Cell Death Differ.. 6. 463-470 (1999)
• [Publications] Mukasa T. et al.: "Activation of caspase-3 apoptotic pathways in skeletal muscle fibers in laminin alpha2-deficient mice"Biochem Biophys Res Commun.. 260. 139-142 (1999)
• [Publications] Fujita, E. et al.: "Akt phosphorylation site found in human caspase-9 is absent in mouse caspase-9"Biochem Biophys Res Commun.. 264. 550-555 (1999)
• [Publications] Fujita, E. et al.: "BMP-4 and retinoic acid synergistically induce activation of caspase-9 and cause apoptosis of P19 embryonal carcinoma cells cultured as a monolayer"Cell Death Differ.. 6. 1109-1116 (1999)