A spatial single-cell atlas of the claustro-insular region uncovers key regulators of neuronal identity and excitability

The claustro-insular region is an evolutionarily conserved and extensively interconnected brain area, critical for functions such as attention, cognitive flexibility, interoception, and affective processing. Despite its importance, its cellular composition and organization remain poorly characterized, hindering a comprehensive understanding of the mechanisms underlying its diverse functions. By combining single-cell RNA sequencing and spatial transcriptomics, we created a high-resolution atlas of this region in mice, uncovering distinct neuronal subtypes and unexpected complexity. Leveraging this atlas, we investigated the role of NR4A2, a neuropsychiatric risk factor expressed in several claustro-insular neuronal subtypes. In an Nr4a2 haploinsufficiency model, we found that only claustrum neurons exhibited shifts in molecular identity. This identity shift, which involved the activation of a transcription factor cascade, was associated with alterations in neuronal firing activity. Our findings provide new insights into the cellular architecture of the claustro-insular region and highlights Nr4a2 as a master regulator of its components identities.