• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to project page

1999 Fiscal Year Final Research Report Summary

EXAMINATION OF MECHANISMS OF EARLY PROGRAMMED MOTONEURON DEATH IN THE DEVELOPING CHICK CERVICAL SPINAL CORD

Research Project

Project/Area Number 10680704
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Nerve anatomy/Neuropathology
Research InstitutionFUKUSHIMA MEDICAL UNIVERSITY

Principal Investigator

YAGINUMA Hiroyuki  FUKUSHIMA MEDICAL UNIVERSITY, DEPARTMENT OF ANATOMY, PROFESSOR, 医学部・解剖学第一講座, 教授 (90230193)

Co-Investigator(Kenkyū-buntansha) NISHIYAMA Keiji  FUKUSHIMA MEDICAL UNIVERSITY, DEPARTMENT OF ANATOMY, ASSOCIATE PROFESSOR, 医学部・解剖学第一講座, 助教授 (10106354)
Project Period (FY) 1998 – 1999
Keywordscaspase / DEVD / BAF / chick / motoneuron / cervical / apoptosis / programmed cell death
Research Abstract

In the chick embryo spinal cord, two types of programmed cell death (PCD) of motoneurons (MNs) are known to occur, one at a relatively late stage and the other at an earlier stage of differentiation. In the second type, the PCD of MNs occurs only in the non-limb innervating cervical spinal cord between E4 and E5. To elucidate the roles for caspases in this early PCD of MNs in the cervical spinal cord, we examined the activation of caspases and the effects of inhibitors of caspases in vivo.
Caspase-3 like activity was increased in ventral MN containing region of the cervical segments at st 24 (E4.5) when many pyknotic dying neurons are observed. Immunohistochemical observations with an antibody against activated form of caspase-3 suggested that activation of caspase-3 precedes both DNA fragmentation and the morphological manifestation of PCD in cervical MNs. To further investigate roles for casapase-3 like activity, we treated embryos with Ac-DEVD-CHO that inhibits caspase-3 like activit … More y. In Ac-DEVD-CHO treated embryos, the nuclei of pyknotic cells showed only moderate condensation. When pyknotic cells were counted, including these moderately pyknotic cells, there was no significant difference between controls and Ac-DEVD-CHO treated embryos. TUNEL staining revealed that DNA fragmentation did not occur in these moderately condensed nuclei, even though electron microscopic observations clearly revealed that degenerative changes were occurring in the cytoplasm. These results suggest that caspase-3 like activity plays a role in the apoptotic nuclear changes.
To investigate involvement of other caspases that are responsible for apoptotic changes of cytoplasm, we treated embryos with Boc-Asp-FMK (BAF) that inhibits broad spectrum of caspases. After treatment with BAF for 12 hours the number of pyknotic cells remarkably fewer than controls. Even moderately pyknotic cell that were observed after treatment with Ac-DEVD-CHO were rarely observed. Instead, there were many cells whose nuclei are smaller and whose cytoplasm was slightly more eosinophilic. These cells lacked DNA fragmentation and retained MN specific marker. Electron microscopic observation revealed that there are many smaller cells that have irregular shaped nuclei, slightly electron dense cytoplasm and apparently normal organelles. Following treatment with BAF for 24 hours, the number of pyknotic cells increased to the level comparable to embryos treated with Ac-DEVD-CHO, although the number of TUNEL positive cells was still fewer than controls. Electron microscopic observation revealed that there are many aberrantly degenerating cells. The total number of surviving MNs was not more than controls and Ac-DEVD-CHO treated embryos. These results suggest that other caspases that can be inhibited by BAF but not by Ac-DEVD-CHO play roles in execution of apoptotic changes in dying MNs. Although inhibition of such caspases considerably delayed the rate of PCD, MNs can not be finally rescued by inhibition of caspases. Less

  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] OPPENHEIM RW 他: "Mdulation of early but not later stages of programmed cell death in embryonic avian spinal cord by sonic hedgehog."Mol.Cell.Neurosci.. 13. 348-361 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] LI X-K 他: "Inhibithion of Fas-Mediated fulminant hepatitis in CrmA Gene-Transfected mice."Biochemical and Biophysical Research Communications.. 273. 101-109 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] HOMMA S 他: "Expression pattem of GDNF, c-ret, and GFR s suggests novel roles for GDNF ligands during early organogenesis in the chick embryo."Developmental Biology. 217. 121-137 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] LIU L 他: "Suppressive effects of Neiting acupuncture on toothache : an expenmental analysis on Fos expression evoked by tooth pulp stimulation in the trigeminal subnucleus pars caudalis and the periaqueductal gray of rats."Neuroscience Research. 38. 331-339 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] YAGINUMA H 他: "Caspase activity is involved in, but is dispensable for, early motoneuron death in the chick embryo cervical spinal cord."(投稿中).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 八木沼洋行: "神経系の発生過程に起こるプログラム細胞死."電子顕微鏡. 34. 93-97 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Oppenheim RW 他: "Normal Programmed Cell death of Developing Avian and Mammalian Neurons Following Inhibition or Genetic deletion of Caspases in Neuronal Death, by Accicdent or by Design"Fondation Ipsen (印刷中)(分担). (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 八木沼洋行: "神経系のprogrammed cell deathとアポトーシス.アポトーシスと疾患"医薬ジャーナル社(分担). 251(23-33) (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] OPPENHEIM RW, HOMMA S, MARTI E, PREVETTE D, WANG S, YAGINUMA H, McMAHON AP: "Mdulation of early but not later stages of programmed cell death in embryonic avian spinal cord by sonic hedgehog."Mol.Cell.Neurosci.. 13. 348-361 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] YAGINUMA Hiroyuki, SHIMADA Takako, SHIRAIWA Nobuko. NISHIYAMA Keiji: "Roles of caspases in programmed motoneuron death in the chick embryo."Italian Journal of Anatomy and Embryology. 104(1). 767 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] LIX-K, FUJINO M, GUO L, OKUYAMA T, FUNESHIMA N, HASHIMOTO M, OKABE K, YAGINUMA H, MIKOSHIBA K, ENOSAWA S, AMEMIYA H, SUZUKI S: "Inhibithion of Fas-Mediated fulminant hepatitis in CrmA Gene-Transfected mice."Biochemical and Biophysical Research Communications.. 273. 101-109 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] HOMMA A, OPPENHEIM RW, YAGINUMA H, KIMURA S: "Expression pattern of GDNF, c-ret, and GFRs suggests novel roles for GDNF ligands during early organogenesis in the chick embryo."Developmental Biology. 217. 121-137 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] LIU L, NISHIYAMA K, HONDA T, SUGIURA M, YAGINUMA H, SUGIURA Y: "Suppressive effects of Neiting acupuncture on toothache : an experimental analysis on Fos expression evoked by tooth pulp stimulation in the trigeminal subnucleus pars caudalis and the periaqueductal gray of rats."Neuroscience Research. 38. 331-339 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] OPPENHEIM RW, KUAN C-Y, PREVETTE D, RAKIC P, YAGINUMA H: "Normal Programmed Cell death of Developing Avian and Mammalian Neurons Following Inhibition or Genetic deletion of Caspases. in Neuronal Death, by Accicdent of by Design."(in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] YAGINUMA H, SHIRAIWA N, SHIMADA T, NISHIYAMA K, HONG J, WANG S, MOMOI T, UCHITAMA Y, OPPENHEIM RW: "Caspase activity is involved in, but is dispensable for, early motoneuron death in the chick embryo cervical spinal cord."(submitted).

    • Description
      「研究成果報告書概要(欧文)」より

URL: 

Published: 2002-03-26  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi