Department of Genetics and Genome Sciences, University of Connecticut School of Medicine, 263 Farmington Avenue, Farmington, CT 06030-6403, USA.
Institute for Systems Genomics, University of Connecticut, 400 Farmington Avenue, Farmington, CT 06030-6403, USA.
Sci Adv. 2022 Apr;8(13):eabl9156. doi: 10.1126/sciadv.abl9156. Epub 2022 Apr 1.
Recent studies using single-cell RNA-sequencing have revealed cellular heterogeneity in the developing mammalian cerebellum, yet the regulatory logic underlying this cellular diversity remains to be elucidated. Using integrated single-cell RNA and ATAC analyses, we resolved developmental trajectories of cerebellar progenitors and identified putative trans- and cis-elements that control cell state transition. We reverse engineered gene regulatory networks (GRNs) of each cerebellar cell type. Through in silico simulations and in vivo experiments, we validated the efficacy of GRN analyses and uncovered the molecular control of a posterior transitory zone (PTZ), a distinct progenitor zone residing immediately anterior to the morphologically defined rhombic lip (RL). We showed that perturbing cell fate specification in the PTZ and RL causes posterior cerebellar vermis hypoplasia, the most common cerebellar birth defect in humans. Our study provides a foundation for comprehensive studies of developmental programs of the mammalian cerebellum.
最近使用单细胞 RNA 测序的研究揭示了哺乳动物小脑发育过程中的细胞异质性，但控制这种细胞多样性的调控逻辑仍有待阐明。我们使用整合的单细胞 RNA 和 ATAC 分析，解析了小脑祖细胞的发育轨迹，并确定了控制细胞状态转变的潜在顺式和反式元件。我们对每个小脑细胞类型的基因调控网络 (GRN) 进行了反向工程。通过计算机模拟和体内实验，我们验证了 GRN 分析的有效性，并揭示了一个独特的前体细胞区（PTZ）的分子调控，该区域位于形态定义的菱形唇（RL）的前部。我们表明，在 PTZ 和 RL 中干扰细胞命运特化会导致小脑蚓部后叶发育不良，这是人类最常见的小脑出生缺陷。我们的研究为全面研究哺乳动物小脑的发育程序提供了基础。
