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Trial of regeneration of retinal neuronal cells by myoseverin

Myoseverin 视网膜神经元细胞再生试验

基本信息

批准号：
13671830
负责人：
IWAMOTO Takashi
金额：
$ 2.24万
依托单位：
Nagoya 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-13671830/
关键词：
myoseverin PC12 retinal pigment epithelial cell differentiation regeneration NGF bFGF 神経成長因子 RalGDS

项目摘要

Myoseverin is a microtubule-binding molecule that induces the dedifferentiation of C2C12 myotubes to individual muscle cells. Here we present the trial of the neural cell regeneration by myoseverin.1) Rat pheochromocytoma PC12 cells differentiate to nerve cells by nerve growth factor (NGF). Addition of myoseverin did not promote of dedifferentiation of NGF-treated PC12 cells. However, when myoseverin was added simultaneously with NGF to PC12 cells, extension of axons was inhibited in about 15% cells. But they did not proliferate and finally exhibited apoptic cell death.2) The induction of RalGDS has been known to inhibit the NGF-induced differentiation of PC12 cells. We found that myoseverin induced the mRNA of RalGDS in PC12 cells. Thus, RalGDS might play an important role in inhibiting NGF-induced differentiation.3) We tried to clone the additional myoseverin-induced genes in PC12 cells and ribosomal PO gene, cofilin and GADD45β were cloned. We established the individual PC12 cell lines that stably expressed those genes and examined their response to NGF. Those cell lines, however, differentiated upon NGF stimulation as parental PC12 cells.4) Retinal pigment epithelial (RPE) cells of rat embryo at day 12 of gestation were cultured in vitro. Basic fibroblast growth factor (bFGF) induced cell proliferation, suppressed their pigmentation, and increased the expression of CHX10 and nestin that were molecular markers of neural progenitors. Myoseverin enhanced these bFGF effects up to about 30%.5) Although bFGF induced the expression of glial fibrillary acidic protein (GFAP) that was a molecular marker of Muller glial cells after 7 days culture, addition of myoseverin inhibited the induction.Thus, while myoseverin has a cooperative effect on the transdifferentiation of RPE induced by bFGF during an initial stage, probably its toxicity inhibited their further differentiation to mature neural cells.

Myoseverin是一种微管结合分子，可诱导C2C12肌管去分化为单个肌细胞。1）神经生长因子（Nerve Growth Factor，NGF）诱导大鼠嗜铬细胞瘤PC12细胞向神经细胞分化。添加myoseverin没有促进NGF处理的PC12细胞的去分化。然而，当将myoseverin与NGF同时加入到PC12细胞中时，约15%的细胞中轴突的延伸被抑制。2）RalGDS的诱导可以抑制NGF诱导的PC12细胞的分化。我们发现myoseverin诱导PC12细胞中RalGDS的mRNA。因此，RalGDS可能在抑制NGF诱导的分化中发挥重要作用。3）我们尝试克隆了PC12细胞中myoseverin诱导的额外基因，克隆了核糖体PO基因、cofilin基因和GADD 45 β基因。我们建立了稳定表达这些基因的PC12细胞系，并检测了它们对NGF的反应。4）体外培养孕12天大鼠胚胎视网膜色素上皮（RPE）细胞。碱性成纤维细胞生长因子（bFGF）诱导细胞增殖，抑制其色素沉着，并增加CHX10和巢蛋白的表达，神经祖细胞的分子标记。Myoseverin可增强bFGF的上述作用达30%左右。5）虽然bFGF在培养7天后诱导Muller胶质细胞的分子标志物胶质细胞酸性蛋白（GFAP）的表达，但Myoseverin的加入抑制了这种诱导，因此，虽然Myoseverin在bFGF诱导的RPE转分化的初始阶段具有协同作用，其毒性可能会抑制它们进一步分化为成熟的神经细胞。