Gene expression profile at single cell level from rhesus macaque fetal brain prefrontal cortex and human organotypic dlPFC slices.

The dorsolateral prefrontal cortex (dlPFC) is a derived cortical area in primates that is involved in myriad high-cognitive functions and is associated with several neuropsychiatric disorders. Here, we performed patch-seq and single-nucleus multiomic analyses of the rhesus macaque dlPFC to identify genes governing neuronal maturation during midfetal to late-fetal development. We also employed patch-seq analysis to assess the impact of CHD8 knockdown on human midfetal neuronal development with human organotypic dlPFC slices. Nuclei were isolated from the prefrontal cortex of rhesus macaque fetal brains and subsequently subjected to single-nuclear ATAC-seq(snATAC-seq) and single-nuclear RNA-seq(snRNA-seq) using the 10X Genomics platform. The rhesus macaque acute slice cells obtained from the prefrontal cortex of the fetal brain were patched, and the nucleus of each cell was subsequently isolated for single-nuclear SAMRT-seq(patch-seq). Also, We performed patch-seq on cells from human organotypic dlPFC slices that contained both CHD8 knockdown(infected by lentivirus expressing both mScarlet and shRNA against CHD8) and control cells(infected by lentivirus expressing both mNeon and control shRNA).

背外侧前额叶皮层（dorsolateral prefrontal cortex, dlPFC）是灵长类动物中演化而来的特化皮层区域，参与诸多高级认知功能，并与多种神经精神疾病密切相关。本研究针对恒河猴的dlPFC开展了Patch-seq与单细胞核多组学分析，以鉴定中胎期至晚胎期神经元成熟过程中的调控基因；同时，借助人类器官型dlPFC脑片，通过Patch-seq分析评估了CHD8基因敲低对人类中胎期神经元发育的影响。研究人员从恒河猴胎儿大脑的前额叶皮层分离细胞核，随后依托10X Genomics平台完成单细胞核转座酶可及性测序（single-nuclear ATAC-seq, snATAC-seq）与单细胞核RNA测序（single-nuclear RNA-seq, snRNA-seq）。此外，研究人员对从胎儿大脑前额叶皮层获取的恒河猴急性脑片细胞进行膜片钳记录，分离每个细胞的细胞核并开展单细胞核SAMRT-seq（即Patch-seq）。本研究还对人类器官型dlPFC脑片的细胞开展Patch-seq分析，其中敲低组细胞感染了同时表达mScarlet与靶向CHD8的短发夹RNA（shRNA）的慢病毒，对照组细胞感染了同时表达mNeon与阴性对照shRNA的慢病毒。