CELF2 Deficiency Demonstrates Autism-like Behaviors and Interferes with Late Development of Cortical Neurons in Mice

Purpose: Celf2 variants have been associated with neurodevelopmental disorders (NDD) including Autism Spectrum Disorders (ASD). This study aims to explore the role of CELF2 in ASD and related NDDs.Methods: Behavioral tests were performed to delineate the behavioral phenotype of celf2 nestin-cre conditional knock out mice. The cerebral cortex of mice on postnatal day 0 was sectioned and stained to evaluate cortical development. In vitro, the neuronal development was assessed by culturing primary neurons of mice with celf2 knockdown, while in vivo, the development and synaptic maturation of cortical neurons in heterozygous knockout mice were examined by Golgi staining. Finally, RNA-seq was employed to identify CELF2 targets and explore the potential mechanism.Results: Our findings revealed that celf2 nestin-cre heterozygous knockout mice exhibited social impairment and anxiety, an autism-like behavior, though no manifestations of repetitive stereotyped behavior, learning cognitive impairment, or depression were observed. Immunofluorescence assay showed an underdeveloped cerebral cortex with significantly reduced cortical thickness, albeit without abnormal cell density. Further in vitro neuronal culture demonstrated a significant reduction in dendritic spines density and affected synaptic maturation in celf2 deficient mice, with no notable abnormalities in total neurite and axon length. RNA-seq analysis of the cerebral cortex revealed differentially expressed genes post celf2 gene knockout compared with the control group. Enrichment analysis highlighted significant enrichment in dendritic and synapse-related biological processes and pathways.Conclusions: Our study delineated the behavioral and neurodevelopmental phenotypes of celf2, suggesting its potential involvement in autism through the regulation of target genes associated with dendritic spines and synapse development. Further research is needed to elucidate the specific mechanisms involved.