Transcriptional profile changes in dedifferentiating Müller glia in mammalian cell culture after treatment with an α7 nicotinic acetylcholine receptor agonist

Conclusions: PNU-282987 activation of RPE causes signficant transcript profile changes in MG cell culture that leads to upregulation of the HB-EGF/Ascl1/Lin28 pathway that was seen with RNA-seq profiles and validated with qRT-PCR. Further, these gene expression changes lead to dedifferentation of MG to an RPC-like fate and expression of several retinal RPC markers were found in RNA-seq data set and was validated with immunocytochemistry. Our study is the first to show that activaiton of the α7 nAChR on RPE can lead to MG dedifferentiation and that the HB-EGF/Ascl1/Lin28 pathway is upregulated in these cells.结论：PNU-282987激活视网膜色素上皮（retinal pigment epithelium, RPE）细胞后，可使穆勒胶质细胞（Müller glia, MG）的体外培养体系出现显著的转录组谱改变，进而上调HB-EGF/Ascl1/Lin28通路，该结果通过RNA测序（RNA-seq）谱分析得到，并经实时定量聚合酶链反应（qRT-PCR）验证。进一步研究发现，上述基因表达变化可诱导穆勒胶质细胞去分化为视网膜祖细胞（retinal progenitor cell, RPC）样表型，RNA测序数据集中共检测到多种视网膜祖细胞标志物的表达，并通过免疫细胞化学实验得到验证。本研究首次证实，激活视网膜色素上皮细胞上的α7烟碱型乙酰胆碱受体（α7 nicotinic acetylcholine receptor, α7 nAChR）可诱导穆勒胶质细胞去分化，且此类细胞中的HB-EGF/Ascl1/Lin28通路呈现上调表达。