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The hypothalamic suprachiasmatic (SCN) clock contains several neurochemically defined cell groups that contribute to the genesis of circadian rhythms. Using cell-specific and genetically targeted approaches we have confirmed an indispensable role for vasoactive intestinal polypeptide-expressing SCN (SCN VIP ) neurons, including their molecular clock, in generating the mammalian locomotor activity (LMA) circadian rhythm. Optogenetic-assisted circuit mapping revealed functional, di-synaptic connectivity between SCN VIP  neurons and dorsomedial hypothalamic neurons, providing a circuit substrate by which SCN VIP  neurons may regulate LMA rhythms. In vivo photometry revealed that while SCN VIP  neurons are acutely responsive to light, their activity is otherwise behavioral state invariant. Single-nuclei RNA-sequencing revealed that SCN VIP  neurons comprise two transcriptionally distinct subtypes, including putative pacemaker and non-pacemaker populations. Altogether, our work establishes necessity of SCN VIP  neurons for the LMA circadian rhythm, elucidates organization of circadian outflow from and modulatory input to SCN VIP  cells, and demonstrates a subpopulation-level molecular heterogeneity that suggests distinct functions for specific SCN VIP  subtypes.

Suprachiasmatic VIP neurons are required for normal circadian rhythmicity and comprised of molecularly distinct subpopulations