Molecular-biological Research for Neuron-Glial Interaction during retinal regeneration.

视网膜再生过程中神经元-胶质细胞相互作用的分子生物学研究。

• 批准号: 13671854
• 负责人: KITANO Shigehiko
• 金额: $ 2.05万
• 依托单位: Tokyo Women's Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671854/
• 关键词: Retinal Ganglion Cells; Glial cells; Retinal regeneration; Neurotrophin; Cytokine; ニューロトロフィン; 虚血; 興奮性アミノ酸; 一酸化窒素合成酵素

The cytotoxic effects of hypoxia and excitatory amino acids on cultured retinal ganglion cells (RGCs) were studied. And the influence of coculture with retinal Muller glia and cortical astrocytes on cell survival was evaluated. The results from our culture model demonstrate the importance of the neuron-glial interaction in determining cellular viability. Glial cells seem to play an important role in the response to hypoxia and excitotoxicity.Nitric oxide may mediate hypoxic and excitotoxic effects on central neurons. In this culture, the glial cell monolayer is bNOS positive and iNOS negative immunohistochemically. The effect of a nitric oxide synthase (NOS) inhibitor, Nω-nitro-L-arginine (NA), on survival of culture rat retinal ganglion cells subjected to hypoxia and excitotoxicity were investigated. The survival rate of cultured RGCs exposed to hypoxia and excitatory amino acids increased in a dose-dependent fashion with NA. The time course of the survival rate of RGC cultures pretreated with NA showed a better survival rate. A NOS inhibitor provides partial protection of RGCs against hypoxia and excitotoxicity in cell culture.In this culture, the glial cell monolayer express immunocytochemically nerve growth factor (NGF) and brain-derived neuritrophic factor (BDNF), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), transforming growth factor β1 (TGF β1).The ciliary marginal zone (CMZ) of the vertebrate retina contains undifferentiated progenitor cells that continue to proliferate and differentiate into retinal-specific cell types. With this culture system, cells from the CMZ of neonatal rat retina identified as various retinal cell types. To investigate whether conditioned medium with NGF and BDNF can induce differentiation of the cells from the CMZ, however, cells cultured in the conditioned medium showed very little retinal cell differentiation.

研究了缺氧和兴奋性氨基酸对培养视网膜神经节细胞的细胞毒作用。观察视网膜Muller胶质细胞和皮质星形胶质细胞共培养对细胞存活的影响。我们的培养模型的结果证明了神经元-胶质相互作用在决定细胞活力方面的重要性。神经胶质细胞似乎在对缺氧和兴奋性毒性的反应中起重要作用。一氧化氮可能介导中枢神经元的缺氧和兴奋毒性作用。在这种培养中，胶质细胞单层免疫组织化学为bNOS阳性和iNOS阴性。研究了一氧化氮合酶（NOS）抑制剂n ω-硝基- l -精氨酸（NA）对缺氧和兴奋性中毒大鼠视网膜神经节细胞存活的影响。暴露于缺氧和兴奋性氨基酸的RGCs的存活率随NA呈剂量依赖性增加。NA预处理的RGC培养物的存活率时间过程显示出较好的存活率。NOS抑制剂在细胞培养中对RGCs的缺氧和兴奋性毒性提供部分保护。在这种培养中，胶质细胞单层免疫细胞化学表达神经生长因子（NGF）和脑源性神经营养因子（BDNF）、血管内皮生长因子（VEGF）、碱性成纤维细胞生长因子（bFGF）、转化生长因子β1 （TGF β1）。脊椎动物视网膜的睫状体边缘区（CMZ）含有未分化的祖细胞，这些细胞继续增殖并分化为视网膜特异性细胞类型。利用该培养系统，新生大鼠视网膜CMZ细胞被鉴定为多种视网膜细胞类型。为了研究含有NGF和BDNF的条件培养基是否可以诱导CMZ细胞分化，然而，条件培养基中培养的细胞几乎没有视网膜细胞分化。