Generation of cholinergic neuron-specific transgenic and knockout mouse system
| Project/Area Number |
15300129
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurochemistry/Neuropharmacology
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| Research Institution | Kyoritsu University of Pharmacy (2005)
Tokyo Metropolitan Organization for Medical Research (2003-2004) |
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Principal Investigator |
MISAWA Hidemi Kyoritsu University of Pharmacy, Pharmacology, Associate Professor, 薬学部, 助教授 (80219617)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 2005: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2003: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | cholinergic neurons / motor neurons / knockout mouse / oxidative stress / superoxide dismutase / mitochondria / neurodegeneration / Cre recombinase / トランスジェニックマウス / 遺伝子改変 / アセチルコリン / トランスポーター |
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| Research Abstract |
The cholinergic gene locus (CGL) consists of the genes encoding the choline acetyltransferase (ChAT) and the vesicular acetylcholine transporter (VAChT). To establish a cholinergic-specific Cre expressing mouse, we constructed a transgene expression vector (VAChT-Cre) with 11.3 kb human CGL in which a Cre-IRES-EGFP unit was inserted in the VAChT open reading frame. The activity of Cre, whose expression was driven by the VAChT promoter, was examined by crossing a reporter mouse (CAG-CAT-Z), in which expression of LacZ is activated upon Cre-mediated recombination. Transgenic lines with the VAChT-Cre construct displayed the restricted Cre expression in a subset of cholinergic neurons in the somatomotor nuclei and medial habenular nucleus, but absent in visceromotor and other central and peripheral cholinergic neurons. Cre expression was first observed at postnatal day 7 and later detected in approximately 40-60% of somatomotor neurons. Based on the onset of Cre expression, we generated tw
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o mouse lines (two alleles ; VAChT-Cre.Fast and VAChT-Cre.Slow) in which Cre expression reaches to the maximal levels fast and slow, respectively. Mitochondrial dysfunction and oxidative damage are implicated in the pathogenesis of neurodegenerative disease. Mice deficient in the mitochondrial form of superoxide dismutase (SOD2) die during embryonic or early postnatal development, precluding analysis of a pathological role for superoxide in adult tissue. We generated postnatal motor neuron-specific SOD2 knockouts by crossing mice with foxed SOD2 alleles to VAChT-Cre mice. SOD2 immunoreactivity was specifically lost in a subset of somatomotor neurons resulting in enhanced superoxide production. Yet extensive histological examination revealed no signs of oxidative damage in animals up to 1 year after birth. However, disorganization of distal nerve axons following injury was accelerated in SOD2-deficient motor neurons. These data demonstrate that postnatal motor neurons are surprisingly resistant to oxidative damage from mitochondrial-derived superoxide radicals unless stressed. Less
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Report
(4 results)
Research Products
(25 results)
