CRISPR/Cas9 mediated intersectional knockout of GSK3β in D2 receptor expressing mPFC neurons reveals contributions to emotional regulation

BackgroundGlycogen synthase kinase 3{beta} (GSK3{beta}) regulates neurodevelopment, synaptic plasticity as well as mood, cognition, social interaction, and depressive-like behaviors. Inhibition of GSK3{beta} is a shared consequence of treatment by lithium, SSRIs, ketamine and antipsychotics. GSK3{beta} activity is regulated by dopamine D2 receptor signaling and can be inhibited by psychoactive drugs in a D2 receptor dependent manner. Functions of GSK3{beta} in striatal D2 neurons has been studied extensively. However, GSK3{beta} is ubiquitously expressed in the brain and D2 receptor expressing cells are distributed as a mosaic in multiple cortical regions. This complicates the interrogation of GSK3{beta} functions in cortical D2 cells in a circuit defined manner using conventional animal models.\n\nMethodsWe have used a CRISPR/Cas9 mediated intersectional approach to achieve targeted deletion of GSK3{beta} in D2 expressing neurons of the adult medial prefrontal cortex (mPFC).\n\nResultsIsolation and analysis of ribosome associated RNA specifically from mPFC D2 neurons lacking GSK3{beta} demonstrated large scale translatome alterations. Deletion of GSK3{beta} in mPFC D2 neurons revealed its contribution to anxiety-related, cognitive, and social behaviors.\n\nConclusionsOur results underscore the viability of intersectional knockout approach to study functions of a ubiquitous gene in a network defined fashion while uncovering a contribution of GSK3{beta} expressed in mPFC D2 neurons in the regulation of behavioral dimensions related to mood and emotions. This advances our understanding of GSK3{beta} action at a brain circuit level and can potentially lead to the development of circuit selective therapeutics.