Title: Identification of midfetal and late-fetal neuronal maturation-related genes using Patch-seq
Legend: (A) Schematic representation of the Patch-seq experimental design for analyzing midfetal and late-fetal rhesus macaque dlPFC cells. (B) Integration of the 256 cells profiled by Patch-seq onto the snRNA-seq reference UMAP. (C) Pseudotime trajectory based on the gene expression profiles of patched IT neurons from 16 individual rhesus macaque fetal brains at 9 distinct fetal ages, with representative electrophysiological sample traces (voltage responses to depolarizing and hyperpolarizing current injections, and inward sodium currents). (D) In situ hybridization detection and quantification (right panel) of CAMK2A, MICAL2, and RAPGEF4 in PCD100 and PCD155 dlPFC. Scale bar, 20 µm. (****p < 0.0001, t-test). (E-L) Plots illustrating the developmental changes in electrophysiological properties, including membrane capacitance (Cm, E), input resistance (Rin, F), resting membrane potential (RMP, G), and inward sodium current (INa, H) of IT neurons across six different age groups. Bottom panels show the expression levels of Cm-correlated genes (I), Rin-correlated genes (J), RMP-correlated genes (K), and INa-correlated genes (L) in IT neurons across the same age groups. (**p < 0.01, ***p < 0.001, ****p < 0.0001, one-way ANOVA, Tukey’s multiple comparisons test).
Citation: Gao Y, Dong Q, Arachchilage KH, Risgaard R, Sheng J, Syed M, Schmidt DK, Jin T, Liu S, Knaack SA, Doherty D, Glass I, Levine JE, Wang D, Chang Q, Zhao X, Sousa AM. Multimodal analyses reveal genes driving electrophysiological maturation of neurons in the primate prefrontal cortex. bioRxiv [Preprint]. 2024 May 16:2023.06.02.543460. doi: 10.1101/2023.06.02.543460. PMID: 37398253; PMCID: PMC10312516.
Abstract: The prefrontal cortex (PFC) is critical for myriad high-cognitive functions and is associated with several neuropsychiatric disorders. Here, using Patch-seq and single-nucleus multiomic analyses, we identified genes and regulatory networks governing the maturation of distinct neuronal populations in the PFC of rhesus macaque. We discovered that specific electrophysiological properties exhibited distinct maturational kinetics and identified key genes underlying these properties. We unveiled that RAPGEF4 is important for the maturation of resting membrane potential and inward sodium current in both macaque and human. We demonstrated that knockdown of CHD8, a high-confidence autism risk gene, in human and macaque organotypic slices led to impaired maturation, via downregulation of key genes, including RAPGEF4. Restoring the expression of RAPGEF4 rescued the proper electrophysiological maturation of CHD8-deficient neurons. Our study revealed regulators of neuronal maturation during a critical period of PFC development in primates and implicated such regulators in molecular processes underlying autism.
Investigator: André Sousa, PhD
About the Lab: The Sousa lab aims to identify and characterize the molecular and cellular mechanisms that govern human brain development and evolution, and to apply that knowledge towards understanding neurodevelopmental and psychiatric disorders.