Department of Biological Structure, University of Washington, Seattle, WA 98195, USA; Department of Pathology, Molecular Medicine and Mechanisms of Disease Program, University of Washington, Seattle, WA 98195, USA.
Department of Biological Structure, University of Washington, Seattle, WA 98195, USA.
Cell Rep. 2020 Feb 18;30(7):2195-2208.e5. doi: 10.1016/j.celrep.2020.01.075.
Müller glia (MG) serve as sources for retinal regeneration in non-mammalian vertebrates. We find that this process can be induced in mouse MG, after injury, by transgenic expression of the proneural transcription factor Ascl1 and the HDAC inhibitor TSA. However, new neurons are generated only from a subset of MG. Identifying factors that limit Ascl1-mediated MG reprogramming could make this process more efficient. In this study, we test whether injury-induced STAT activation hampers the ability of Ascl1 to reprogram MG into retinal neurons. Single-cell RNA-seq shows that progenitor-like cells derived from Ascl1-expressing MG have a higher level of STAT signaling than do those cells that become neurons. Ascl1-ChIPseq and ATAC-seq show that STAT potentially directs Ascl1 to developmentally inappropriate targets. Using a STAT inhibitor, in combination with our previously described reprogramming paradigm, we found a large increase in the ability of MG to generate neurons.
Müller 胶质细胞 (MG) 是无脊椎动物视网膜再生的来源。我们发现，在小鼠 MG 受到损伤后，通过过表达神经前体细胞转录因子 Ascl1 和组蛋白去乙酰化酶抑制剂 TSA，可诱导这一过程。然而，新的神经元仅由 MG 的一部分产生。确定限制 Ascl1 介导的 MG 重编程的因素，可以使这一过程更加高效。在这项研究中，我们测试了损伤诱导的 STAT 激活是否会阻碍 Ascl1 将 MG 重编程为视网膜神经元的能力。单细胞 RNA-seq 表明，与那些成为神经元的细胞相比，源自表达 Ascl1 的 MG 的祖细胞样细胞具有更高水平的 STAT 信号。Ascl1-ChIPseq 和 ATAC-seq 表明，STAT 可能将 Ascl1 导向发育不当的靶标。使用 STAT 抑制剂，结合我们之前描述的重编程范例，我们发现 MG 生成神经元的能力大大提高。
