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

• 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
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591848/
• 关键词: retinal ganglion cell; retinal glial cell; culture experiment; cellular interaction; DNA chip; nitric oxide; gravity loading; ミュラー細胞; 視神経アストロサイト; 緑内障; 虚血; Microarray; Differential display; 軸索流

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.

体外结果在体外结果不一致并不罕见。培养细胞与体内的环境差异应该是可能的原因之一。但是，用其他类型的细胞培养的细胞不适合研究其针对某些负载的纯反应。在这项研究中，我们采用了纯粹的分离细胞来识别其针对某些负载的特定反应，并使用不同类型的纯粹孤立细胞建立了一些人工重建系统，以清楚地研究它们的相互作用。总之，它能够分析单型细胞反应针对几个负载条件和分子水平的细胞间反应。研究纯粹分离的细胞对几个负载条件的反应，我们开发了一种新的定量重力加载系统。我们研究了纯粹分离的视网膜神经节细胞（RGC）和视网膜神经胶质细胞之间的差异，以针对其存活率和使用DNA芯片法的mRNA反应中的重力负载。在结果中，两种细胞类型之间的加载反应并不相同。我们还揭示了RGC和视网膜神经胶质细胞之间针对卫生负荷产生的一氧化氮速率不同。我们使用纯粹分离的RGC和视网膜细胞重建了人工混合培养条件。在结果中，视网膜神经胶质细胞主要反应应力载荷，导致对RGC存活的继发作用。这些结果表明，研究神经保护和神经元死亡机制时，研究细胞间反应非常重要。

项目成果

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)。

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

先端医療シリーズ23・眼科 眼科の最新医療

高级医学系列23/眼科 眼科最新医疗护理

• 发表时间: 2003
• 作者: Kashiwagi;K;粕木賢治
• 通讯作者: 粕木賢治

Molecular and cellular reactions of retinal ganglion cells and retinal glial cells under centrifugal force loading

• DOI: 10.1167/iovs.04-0277
• 发表时间: 2004-10-01
• 期刊: INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
• 影响因子: 4.4
• 作者: Kashiwagi, K;Iizuka, Y;Tsukahara, S
• 通讯作者: Tsukahara, S

柏木賢治: "Increase in Dephosphorylation of the Heavy Neurofilament Subunit in the Monkey Chronic Glaucoma Model"INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE. 44(1). 154-159 (2003)

Kenji Kashiwagi：“猴子慢性青光眼模型中重神经丝亚基的去磷酸化增加”调查眼科与视觉科学 44（1）（2003）。