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Investigation for mechanism and protection of retinal ganglion cell death using several culture disease-model systems

使用几种培养疾病模型系统研究视网膜神经节细胞死亡的机制和保护

基本信息

批准号：
15591848
负责人：
KASHIWAGI Kenji
金额：
$ 2.11万
依托单位：
University of Yamanashi
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

It is not rare that in vitro results are not consistent in vitro results. The environmental difference of cultured cells from those of in vivo should be one possible reason. However, cells cultured with other types of cells are not suitable for researching their pure reaction against certain loadings. In this study, we employed purely isolated cells for identifying their specific reaction against certain loadings and we established some artificial reconstruction systems using different types of purely isolated cell for investigating their interaction clearly. In conclusion, it is able to analyze single type cell reaction against several loading conditions and intercellular reaction at the molecular level.For studying reactions of purely isolated cells against several loading conditions we developed a novel quantitative gravity loading system. We investigated differences between purely isolated retinal ganglion cell (RGC) and retinal glial cell against gravity loading in their survival rate and in mRNA reaction employing DNA chip method. In results, the reactions to loading were not the same between two cell types. We also revealed that nitric oxide producing rate against hyoxic loading are different between RGC and retinal glial cells. We reconstructed artificial mixed culture condition using purely isolated RGC and retinal glial cells. In results, retinal glial cells primary reacted stress loading resulting secondary effect on RGC survival. These results indicate that it is extremely important to investigate intercellular reaction when investigating neuroprotection and neuronal death mechanism.

体外结果与体外结果不一致的情况并不少见。培养细胞与体内细胞的环境差异应该是一个可能的原因。然而，用其他类型的细胞培养的细胞不适合研究它们对某些负载的纯反应。在这项研究中，我们使用纯分离的细胞来鉴定它们对特定负荷的特异性反应，并建立了一些使用不同类型的纯分离细胞的人工重建系统，以清楚地研究它们之间的相互作用。综上所述，它能够在分子水平上分析单个细胞对几种加载条件的反应以及细胞间的反应。为了研究纯分离细胞对几种加载条件的反应，我们开发了一种新型的定量重力加载系统。采用DNA芯片技术，研究了重力作用下视网膜神经节细胞(RGC)和视网膜神经胶质细胞在存活率和mRNA反应方面的差异。结果表明，两种细胞类型对负载的反应不同。我们还发现RGC和视网膜神经胶质细胞在抗低氧负荷时产生一氧化氮的速率不同。我们用纯分离的RGC和视网膜神经胶质细胞重建了人工混合培养条件。结果显示，视网膜神经胶质细胞主要反应应激负荷，继而影响RGC的存活率。这些结果表明，在研究神经保护和神经元死亡机制时，研究细胞间反应是非常重要的。

项目成果

期刊论文数量（19）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

柏木賢治: "Differences in nitric oxide production : a comparison of retinal ganglion cells and retinal glial cells cultured under hypoxic conditions"MOLECULAR BRAIN RESEARCH. 112(12). 126-134 (2003)

Kenji Kashiwagi：“一氧化氮生成的差异：缺氧条件下培养的视网膜神经节细胞和视网膜神经胶质细胞的比较”MOLECULAR BRAIN RESEARCH 112(12)。

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Neuroprotective effects of erythropoietin on glutamate and nitric oxide toxicity in primary cultured retinal ganglion cells

DOI：
10.1016/j.brainres.2005.05.037
发表时间：
2005-07
期刊：
Brain Research
影响因子：
2.9
作者：
Makiko Yamasaki;H. Mishima;H. Yamashita;K. Kashiwagi;K. Murata;A. Minamoto;T. Inaba
通讯作者：
Makiko Yamasaki;H. Mishima;H. Yamashita;K. Kashiwagi;K. Murata;A. Minamoto;T. Inaba

Increase in dephosphorylation of the heavy neurofilament subunit in the monkey chronic glaucoma model