小鼠不同脑区星形胶质细胞小分子诱导转分化和基因表达差异的研究

The Differences of Transdifferentiation Induced by Small Molecules and Gene Expression between Mouse Astrocytes from Different Brain Regions

该文旨在比较小分子化合物诱导小鼠不同脑区星形胶质细胞向神经元转分化的特性,并利用转录组测序技术分析小鼠不同脑区星形胶质细胞的基因表达差异。以新生小鼠皮层和海马的星形胶质细胞作为起始细胞,通过小分子化合物VCR诱导其向神经元转分化,利用免疫荧光染色检测转分化过程中细胞形态的变化以及神经元的比例,通过转录组测序比较两种星形胶质细胞的基因表达差异,并对差异基因进行荧光定量PCR验证及GO富集分析。结果表明,皮层星形胶质细胞经VCR诱导转分化为神经元的能力要显著优于海马星形胶质细胞;转录组测序发现,两种星形胶质细胞有12658个基因存在差异表达,GO分析结果表明,在皮层星形胶质细胞中高表达的基因更多地参与细胞分裂的过程,推测差异显著基因GAD2、EYA2、GSX2、INSM1以及GNG3是与转分化相关的基因。该研究对星形胶质细胞向神经元转分化的机制研究具有借鉴意义。

The current study aimed to compare the neuronal transdifferentiation ability induced by small molecules of astrocytes derived from different mouse brain regions,and analyze the differentially expressed genes in these astrocytes by transcriptome sequencing.Astrocytes isolated from the cortex and hippocampus of neonatal mice were induced into neuronal transdifferentiation by a small molecule cocktail,VCR.The morphological changes of cells and ratio of neurons in total cells in the process of transdifferentiation were investigated by immunofluorescence staining.Then transcriptome sequencing was carried out to compare the differences in gene expression between the two types of astrocytes.The sequencing results were further verified by qPCR and analyzed by GO analysis.The results showed that the transdifferentiation efficiency of cortical astrocytes was significantly higher than that of hippocampal astrocytes.The transcriptome sequencing results revealed that 12658 genes were differentially expressed between the two kinds of astrocytes.GO analysis showed that genes highly expressed in cortical astrocytes were mainly involved in the process of cell division.It is speculated that some key genes,such as GAD2,EYA2,GSX2,INSM1 and GNG3,significantly highly expressed in cortical astrocytes might be related to transdifferentiation.This study provides a reference for further mechanism study of regional specific astrocytes-to-neuron conversion.