1997 Fiscal Year Final Research Report Summary
ANALYSIS OF MECHANISMS OF NEURONAL CELL DEATH USING ADENOVIRUS VECTOR SYSTEM
| Project/Area Number |
08680808
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Nerve anatomy/Neuropathology
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| Research Institution | FUKUSHIMA MEDICAL UNIVERSITY (1997)
University of Tsukuba (1996) |
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Principal Investigator |
YAGINUMA Hiroyuki FUKUSHIMA MEDICAL UNIVERSITY,DEPARTMENT OF ANATOMY, PROFESSOR, 解剖学第一講座, 教授 (90230193)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Bcl-2 / Crm A / Adenovirus / chick / motoneuron / Dorsal root ganglion / apoptosis / programmed cell death |
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| Research Abstract |
Aims of this project were (I) to establish adenovirus vector system that is capable of introducing genes into neuronal cells of the chick embryo, and (II) to analyze effects of Bcl-2 and Crm A on neuronal cell death in vitro and in vivo. Major results are the following :
(1) In the first trial, we failed to obtain recombinant adenovirus that express Bcl-2 under regulation of CAG promoter. Employing Cre-LoxP system that can regulate expression of genes, we succeeded to obtain the recombinant adenovirus that express Bcl-2.
(2) It was confirmed that genes introduced by adenovirus vecotrs were expressed in the chick DRG neurons and chick motoneurons in vitro. Introduced human Bcl-2 could rescue DRG neurons and motoneurons from cell death in vitro.
(3) It is also confirmed that adenovirus vectors injected into leg bud were retrogradely transported to the motoneurons in the lateral motor column. Bcl-2 introduced with this retrograde transport could rescue some motoneurons after limb bud removal.
(4) In the rat C6 glioma cell line system, human Bcl-2 introduced by adenovirus vector induced cell death.
(5) We could not find rescuing effects of Crm A gene introduced by adenovirus vector in the DRG neuron system.
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[Publications] Sat N., Wang S., Li, L.Okabe K., Hashimoto M., Yaginuma, H., Mikoshiba K., Uetsuki T., Yoshikawa K., Milligan C.E., and Oppenheim, R.W.: "A novel strategy for introducing exogenous Bcl-2 into neuronal cells : the Cre-loxP system-mediated activation of Bcl-2 for preventing programmed cell death using recombinant adenoviruses." Mol.Cell.Neurosci.12. 65-78 (1998)
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[Publications] Aruga, J., Minowa, O., Yaginuma, H., Kuno, J., Nagai, T., Noda, T., and Mikoshiba, K.: "Mouse Zic1 is involved in cerebellar development." J.Neurosci.18. 284-293 (1998)
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