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从 Sox10-Venus 小鼠中分化少突前体细胞为少突胶质细胞和星形胶质细胞。

Differentiation of Oligodendrocyte Precursor Cells from Sox10-Venus Mice to Oligodendrocytes and Astrocytes.

机构信息

Department of Biochemistry and Biophysics, Graduate School of Health Care Sciences, Tokyo Medical and Dental University, Tokyo, Japan.

Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University, Tokyo, Japan.

出版信息

Sci Rep. 2017 Oct 26;7(1):14133. doi: 10.1038/s41598-017-14207-0.

Abstract

Oligodendrocytes are well known as myelin-forming cells in the central nervous system (CNS). However, detailed mechanisms of oligodendrocyte differentiation and myelination are poorly understood, particularly due to the difficulty of the purification of murine oligodendrocyte precursor cells (OPCs). We have recently established a transgenic mouse line that expresses a fluorescent protein Venus under the promoter of Sox10, whose expression is restricted to OPCs and oligodendrocytes in the CNS. Here, we have characterized Venus-positive cells from the Sox10-Venus mouse brain for analyzing oligodendrocyte differentiation. We first purified Venus-positive cells from the postnatal day 0-2 brain by flow cytometry. Most of the Venus-positive cells expressed NG2, an OPC marker. After induction of differentiation, an increased population of galactocerebroside-positive oligodendrocytes and decrease of OPCs were observed in the Venus-positive culture. Furthermore, a time-lapse analysis showed that Venus-positive oligodendrocytes dynamically changed their morphology with highly branched cell processes during differentiation. In addition, we found that Venus-positive OPCs were able to differentiate to type II astrocytes. In vivo, OPCs and oligodendrocytes express Venus, and some of astrocytes were positive for Venus in the ventral cortex. Taken together, the Sox10-Venus mouse system is useful for analyzing differentiation and multipotency of murine OPCs.

摘要

少突胶质细胞是中枢神经系统(CNS)中形成髓鞘的细胞,这是众所周知的。然而,少突胶质细胞分化和髓鞘形成的详细机制还了解甚少,特别是由于难以纯化鼠少突胶质前体细胞(OPC)。我们最近建立了一个转基因小鼠品系,其在 Sox10 启动子的控制下表达荧光蛋白 Venus,该蛋白的表达仅限于 CNS 中的 OPC 和少突胶质细胞。在这里,我们对 Sox10-Venus 小鼠大脑中的 Venus 阳性细胞进行了特征分析,以研究少突胶质细胞的分化。我们首先通过流式细胞术从出生后 0-2 天的大脑中纯化 Venus 阳性细胞。大多数 Venus 阳性细胞表达 NG2,这是一种 OPC 标志物。诱导分化后,在 Venus 阳性培养物中观察到 GalC 阳性少突胶质细胞的数量增加,而 OPC 数量减少。此外,延时分析显示 Venus 阳性少突胶质细胞在分化过程中其形态具有高度分支的细胞突起,呈现动态变化。此外,我们发现 Venus 阳性 OPC 能够分化为星形胶质细胞 II 型。在体内,OPC 和少突胶质细胞表达 Venus,腹侧皮质中的一些星形胶质细胞也呈 Venus 阳性。总之,Sox10-Venus 小鼠系统可用于分析鼠 OPC 的分化和多能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/febf/5658394/d5d5bf09d27b/41598_2017_14207_Fig1_HTML.jpg

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