Background: The paraventricular nuclei of the hypothalamus (PVH) are important for both body weight and glucose regulation. We previously demonstrated that vitamin D receptors (VDRs) in the PVH are necessary for glucose regulation in a high-fat fed model but notably, do not affect body weight. Previous work in neuronal populations within the PVH almost always show a body weight phenotype. Thus, the lack of body weight effects in our previous work suggests that VDRs may identify a subpopulation of PVH neurons that have distinct glucose-regulating effects. Thus, we aimed to determine the effect of activation and inhibition of VDR-positive neurons in the PVH. Methods: We used a chemogenetic strategy in transgenic mice containing Cre-recombinase in VDR+ neurons (VDR-Cre) combined with intra-PVH injections of a cre-dependent virus to acutely activate (AAV-hM3D(Gq)) or inhibit (AAV-hM4D(Gi)) VDR-positive PVH neurons when activated with CNO. After a 2 week recovery, mice were fasted for 4 hours, injected with either CNO or saline (counterbalanced design), and then underwent a intraperitoneal (ip) glucose tolerance test. Two weeks later, mice underwent a repeat ip glucose tolerance test with the alternate treatment (CNO or saline). After a four week recovery, mice underwent a similar protocol, but instead of receiving ip glucose, had their food intake measured for 24 hours after a 4 hour fast (each animal serving as its own control with all animals receiving both CNO or vehicle). These experiments were repeated in a separate cohort that was placed on a high-fat diet for 8 weeks prior to their initial intra-PVH injections. Results: Chow-fed mice had no effects of either acute activation (hM3D(Gq)) or inhibition (hM4D(Gi)) of PHV-VDR+ neurons on glucose regulation, food intake, or body weight. Additionally, acute inhibition of PVH-VDR+ neurons with hM4D(Gi) did not cause changes in glucose regulation in male or female mice. Studies with VDR-Cre mice with hM3D(Gq) on high fat diet are still ongoing. Conclusions: Acute activation or inhibition of VDR-positive neurons within the PVH does not seem to have measurable effects on glucose homeostasis. Next steps will be to determine if chronic activation or inhibition of these neurons are important for glucose control.