Multiparametric Assays Capture Sex- and Environment-Dependent Modifiers of Behavioral Phenotypes in Autism Mouse Models

Current phenotyping approaches for murine autism models often focus on one selected behavioral feature, making the translation onto a spectrum of autistic characteristics in humans challenging. Furthermore, sex and environmental factors are rarely considered. Here, we aimed to capture the full spectrum of behavioral manifestations in three autism mouse models to develop a "behavioral fingerprint" that takes environmental and sex influences under consideration. To this end, we employed a wide range of classical standardized behavioral tests; and two multi-parametric behavioral assays: the Live Mouse Tracker and Motion Sequencing (MoSeq), on male and female Shank2, Tsc1 and Purkinje cell specific-Tsc1 mutant mice raised in standard or enriched environments. Our aim was to integrate our high dimensional data into one single platform to classify differences in all experimental groups along dimensions with maximum discriminative power. Multi-parametric behavioral assays enabled far more accurate classification of experimental groups compared to classical tests, and dimensionality reduction analysis demonstrated significant additional gains in classification accuracy, highlighting the presence of sex, environmental and genotype differences in our experimental groups. Together, our results provide a complete phenotypic description of all tested groups, suggesting multi-parametric assays can capture the entire spectrum of the heterogenous phenotype in autism mouse models.