Single cell RNA-seq analysis in mouse embryonic medial ganglionic eminence (MGE) under deletion of Ezh2 gene (10X Genomics Multiome)

Enhancer of zeste homolog 2 (Ezh2) is the methyltransferase component of the Polycomb Repressive Complex 2 (PRC2), which is critical for trimethylation of histone 3 at lysine 27 (H3K27me3) and results in gene repression. Mutations in EZH2 and dysregulation of H3K27me3 can lead to neurodevelopmental abnormalities such as Weaver Syndrome and ataxia-telangiectasia. During cortical neurogenesis, H3K27me3 is a critical epigenetic modification that regulates cellular maturation rate, and in turn, determines when precursor cells exit the cell cycle. Loss of function studies reveal that Ezh2 plays a critical role in epigenetic regulation of neuronal fate and maturation in some brain regions, but a role for Ezh2 in forebrain GABAergic interneurons has not been explored. Here, we removed Ezh2 from the medial ganglionic eminence (MGE) to study its role in interneuron development. WT, Het, and Ezh2 KO mice (Nkx2.1-CreC/+;Ezh2;Ai9F/+) were created by Nkx2.1-CreC/C;Ezh2F/+ males crossed to Ai9F/F;Ezh2F/+ females. Single nuclei from embryonic day 12.5 (e12) and 15.5 (e15) from each genotype were prepared to analyze transcriptome using 10X Genomics Multiome pipeline.zeste增强子同源物2（Enhancer of zeste homolog 2，Ezh2）是多梳抑制复合体2（Polycomb Repressive Complex 2，PRC2）的甲基转移酶组分，其在组蛋白H3赖氨酸27三甲基化（histone 3 at lysine 27，H3K27me3）过程中发挥关键作用，并介导基因阻抑。EZH2基因突变及H3K27me3调控异常可导致韦弗综合征（Weaver Syndrome）、共济失调毛细血管扩张症（ataxia-telangiectasia）等神经发育异常。在皮层神经发生过程中，H3K27me3是一种关键的表观遗传修饰，可调控细胞成熟速率，进而决定前体细胞退出细胞周期的时机。功能丧失性研究表明，Ezh2在部分脑区的神经元命运与成熟的表观遗传调控中发挥关键作用，但目前尚未明确其在前脑γ-氨基丁酸能中间神经元中的功能。本研究通过敲除内侧神经节隆起（medial ganglionic eminence，MGE）中的Ezh2，以探究其在中间神经元发育中的作用。通过将Nkx2.1-CreC/C;Ezh2F/+雄性小鼠与Ai9F/F;Ezh2F/+雌性小鼠杂交，获得了野生型（WT）、杂合型（Het）及Ezh2基因敲除型（Ezh2 KO）小鼠（基因型为Nkx2.1-CreC/+;Ezh2;Ai9F/+）。收集各基因型小鼠胚胎第12.5天（e12）和第15.5天（e15）的单细胞核，采用10×基因组学多组学流程（10X Genomics Multiome pipeline）进行转录组分析。