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Quantitative analysis in neurotrophic factor genes of spinal cord with complementary DNA microarray

互补DNA微阵列定量分析脊髓神经营养因子基因

基本信息

批准号：
18390411
负责人：
BABA Hisatoshi
金额：
$ 9.35万
依托单位：
University of Fukui
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

Chronic mechanical compression of the spinal cord causes neural tissue damage, including loss of anterior horn cells around the level of injury. Exogenous delivery of neurotrophins to neuronal cells could provide neuroprotection to a spinal cord subjected to mechanical injury. We investigated the efficacy of retrograde gene delivery of adenoviral vector (AdV) carrying neurotrophin-3 (NT-3) gene into twy (twy/twy) mouse spinal cord anterior horn neurons with chronic and progressive mechanical compression at C1-C2 level. AdV-NT-3 was used for retrograde delivery via the sternomastoid muscle to the cervical spinal accessory motoneurons in 16-week-old adult twy mice with relatively mild spinal cord compression. Four weeks after the AdV-NT-3 or AdV-beta-galactosidase cDNA (LacZ) as a marker gene injection, the compressed cervical spinal cord was examined histologically, immunohistologically, and by immunoblot analysis. Immunoreactivity to NT-3 was significantly enhanced in the AdV-NT-3-injected twy mice compared with the AdV-LacZ-injected mice. The numbers of anterior horn neurons of Nissl-, choline acetyltransferase (ChAT)-, and trkC-stained and wheat germ agglutinin-horseradish peroxidase(WGA-HRP)-labeled neurons at the spinal cord level with maximum compression were significantly higher in AdV-NT-3-transfected than in AdV-LacZ-transfected twy mice. Retrograde NT-3 gene transfer to twy mouse anterior horn neurons increased neurite axonal length and arborization of WGA-HRP-labeled neurons. Our results suggest that targeted retrograde NT-3 gene delivery is feasible in the intact animal and that it enhances neuronal survival even under chronic mechanical compression of the spinal cord. (c) 2008Wiley-Liss, Inc.

脊髓的慢性机械压迫会导致神经组织损伤，包括损伤水平周围的前角细胞丢失。外源性神经营养因子对神经元细胞的递送可以对遭受机械损伤的脊髓提供神经保护。采用慢性和进行性机械压迫的方法，观察了腺病毒载体（AdV）携带神经营养素-3（NT-3）基因逆行转染twy（twy/twy）小鼠C1-C2脊髓前角神经元的效果。AdV-NT-3用于通过胸锁乳突肌逆行递送至具有相对轻度脊髓压迫的16周龄成年twy小鼠的颈脊髓副运动神经元。4周后，AdV-NT-3或AdV-beta-半乳糖苷酶cDNA（LacZ）作为标记基因注射，压缩颈脊髓进行组织学检查，免疫组织学，并通过免疫印迹分析。与注射AdV-LacZ的小鼠相比，注射AdV-NT-3的twy小鼠对NT-3的免疫反应性显著增强。Nissl染色、胆碱乙酰转移酶（ChAT）染色、trkC染色和麦胚凝集素-辣根过氧化物酶（WGA-HRP）标记的脊髓前角神经元数量在AdV-NT-3转染组显著高于AdV-LacZ转染组。逆行NT-3基因转移到小鼠前角神经元增加轴突长度和WGA-HRP标记的神经元的树状化。我们的研究结果表明，有针对性的逆行NT-3基因传递是可行的，在完整的动物，它提高了神经元的生存，即使在慢性机械压迫脊髓。(c)2008Wiley-Liss，Inc.