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.

体外结果与体外结果不一致的情况并不罕见。培养细胞与体内环境的差异应该是原因之一。然而，与其他类型的细胞一起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，我们采用纯分离的细胞来识别它们对某些负载的特定反应，并使用不同类型的纯分离的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，它能够在分子水平上分析单一类型细胞在多种加载条件下的反应和细胞间反应。为了研究纯分离细胞在多种加载条件下的反应，我们开发了一种新型定量重力加载系统。我们采用 DNA 芯片方法研究了纯分离的视网膜神经节细胞 (RGC) 和视网膜神经胶质细胞在重力负荷下的存活率和 mRNA 反应方面的差异。结果，两种细胞类型对负载的反应并不相同。我们还发现，RGC 和视网膜神经胶质细胞针对缺氧负荷的一氧化氮生成率不同。我们使用纯分离的 RGC 和视网膜神经胶质细胞重建了人工混合培养条件。结果，视网膜神经胶质细胞主要对应激负荷做出反应，从而对 RGC 存活产生继发性影响。这些结果表明，在研究神经保护和神经元死亡机制时，研究细胞间反应极其重要。

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）。