Artegiani Benedetta, de Jesus Domingues Antonio M, Bragado Alonso Sara, Brandl Elisabeth, Massalini Simone, Dahl Andreas, Calegari Federico
DFG-Research Center for Regenerative Therapies, Cluster of Excellence, TU-Dresden, Dresden, Germany.
Deep Sequencing Group-SFB655, Biotechnology Center, TU-Dresden, Dresden, Germany.
EMBO J. 2015 Apr 1;34(7):896-910. doi: 10.15252/embj.201490061. Epub 2014 Dec 19.
Major efforts are invested to characterize the factors controlling the proliferation of neural stem cells. During mammalian corticogenesis, our group has identified a small pool of genes that are transiently downregulated in the switch of neural stem cells to neurogenic division and reinduced in newborn neurons. Among these switch genes, we found Tox, a transcription factor with hitherto uncharacterized roles in the nervous system. Here, we investigated the role of Tox in corticogenesis by characterizing its expression at the tissue, cellular and temporal level. We found that Tox is regulated by calcineurin/Nfat signalling. Moreover, we combined DNA adenine methyltransferase identification (DamID) with deep sequencing to characterize the chromatin binding properties of Tox including its motif and downstream transcriptional targets including Sox2, Tbr2, Prox1 and other key factors. Finally, we manipulated Tox in the developing brain and validated its multiple roles in promoting neural stem cell proliferation and neurite outgrowth of newborn neurons. Our data provide a valuable resource to study the role of Tox in other tissues and highlight a novel key player in brain development.
人们投入了大量精力来确定控制神经干细胞增殖的因素。在哺乳动物皮质发生过程中，我们的研究小组已鉴定出一小部分基因，这些基因在神经干细胞向神经源性分裂转变时会短暂下调，并在新生神经元中重新诱导表达。在这些转换基因中，我们发现了Tox，这是一种在神经系统中作用尚未明确的转录因子。在此，我们通过在组织、细胞和时间水平上表征其表达来研究Tox在皮质发生中的作用。我们发现Tox受钙调神经磷酸酶/Nfat信号传导调控。此外，我们将DNA腺嘌呤甲基转移酶鉴定（DamID）与深度测序相结合，以表征Tox的染色质结合特性，包括其基序以及下游转录靶点，如Sox2、Tbr2、Prox1和其他关键因子。最后，我们在发育中的大脑中对Tox进行操作，并验证了其在促进神经干细胞增殖和新生神经元神经突生长中的多种作用。我们的数据为研究Tox在其他组织中的作用提供了宝贵资源，并突出了其在大脑发育中的一个新的关键作用因子。
