F. Hoffmann-La Roche, 4070 Basel, Switzerland.
Department of Ophthalmology, University Hospital of Bern, University of Bern, 3010 Bern, Switzerland.
Int J Mol Sci. 2019 Jul 19;20(14):3547. doi: 10.3390/ijms20143547.
Lgr5, an intestinal adult stem cell marker, was recently also found in neuronal tissues. We investigated whether retinal Lgr5 cells express properties of neural stem cells (NSC) and/or of differentiated interneurons during retinal development. RNA was isolated from Lgr5 and Lgr5 populations from postnatal day 5 (PN5) and adult retinas of Lgr5 knock-in mice sorted by fluorescence-activated cell sorting (FACS). Transcriptome analyses were performed on two RNA samples of each developmental stage (PN5 and adult). The online platform PANTHER (Protein ANalysis THrough Evolutionary Relationships) was used to determine overrepresented gene ontology (GO) terms of biological processes within the set of differentially expressed genes. The detailed evaluation included gene expression in regard to stem cell maintenance/proliferation, cell cycle, and Wnt signaling but also markers of differentiated retinal neurons. None of the enriched GO terms of upregulated genes of Lgr5 cells showed a positive association to NSC. On the contrary, NSC maintenance and proliferation rather prevail in the Lgr5 cell population. Furthermore, results suggesting that Wnt signaling is not active in the Lgr5 population. Therefore, our transcriptome analysis of Lgr5 retinal cells suggest that these cells are differentiated neurons, specifically glycinergic amacrine cells.
Lgr5 是一种肠道成体干细胞标志物,最近也在神经元组织中被发现。我们研究了视网膜 Lgr5 细胞在视网膜发育过程中是否表达神经干细胞 (NSC) 和/或分化的中间神经元的特性。从小鼠的视网膜中分离出 Lgr5 和 Lgr5 群体的 RNA,这些细胞是通过荧光激活细胞分选 (FACS) 从出生后 5 天 (PN5) 和成年 Lgr5 敲入小鼠的视网膜中分选出来的。对每个发育阶段 (PN5 和成年) 的两个 RNA 样本进行了转录组分析。在线平台 PANTHER(通过进化关系进行蛋白质分析)用于确定差异表达基因集中的生物学过程中过度表达的基因本体 (GO) 术语。详细的评估包括与干细胞维持/增殖、细胞周期和 Wnt 信号有关的基因表达,但也包括分化的视网膜神经元的标志物。Lgr5 细胞上调基因的富集 GO 术语中没有一个与 NSC 呈正相关。相反,Lgr5 细胞群体中更倾向于 NSC 的维持和增殖。此外,结果表明 Wnt 信号在 Lgr5 群体中不活跃。因此,我们对 Lgr5 视网膜细胞的转录组分析表明,这些细胞是分化的神经元,特别是甘氨酸能中间神经元。