Cntn4 Gene Deficiency Promotes Autism-like Phenotypes Associated with Gut Microbiota Perturbations and Gut-Brain Axis Metabolomic Alterations in Mice

Autism spectrum disorder (ASD) has a pathogenesis that involves complex genetic-environmental interactions. To investigate whether the ASD risk gene Cntn4 contributes to disease development via gut-brain axis mechanisms. In this study, a Cntn4 gene knockout mouse model was established. After behavioral tests, 16S rDNA sequencing was employed to evaluate microbial diversity and abundance in the gut contents, while metabolomic methods were applied to detect changes in metabolites in gut contents, serum and cerebral cortex. The results demonstrated that knockdown of the Cntn4 gene induced autism-like behavioral changes in the mice. Significant differences were observed in both intestinal microbiota composition and metabolomic profiles compared to control mice. Arginine and proline metabolism, histidine metabolism, sphingolipid metabolism, tyrosine metabolism, and purine metabolism, were notably affected. Changes in the abundance of certain microbial taxa, such as Adlercreutzia and Desulfovibrionaceae_unclassified, were correlated with alterations in metabolites, particularly organic acids and their derivatives, across different sample types. These findings suggest that knockout of the Cntn4 gene leads to autism-like changes in mice through mechanisms associated with alterations in gut microbiota and gut-brain axis metabolites. This study provides valuable insights into the mechanisms underlying ASD development and offers potential directions for the prevention and treatment of ASD.