To investigate vitamin D-related control of brain-expressed genes, candidate vitamin D responsive elements (VDREs) at 27/210 kb in human tryptophan hydroxylase (TPH)2 were probed. Both VDREs bound the vitamin D receptor (VDR)-retinoid X receptor (RXR) complex and drove reporter gene transcription in response to 1,25-dihydroxyvitamin D 3 (1,25D). Brain TPH2 mRNA, encoding the rate-limiting enzyme in serotonin synthesis, was induced 2.2-fold by 10 nM 1,25D in human U87 glioblastoma cells and 47.8-fold in rat serotonergic RN46A-B14 cells. 1,25D regulation of leptin (Lep), encoding a serotoninlike satiety factor, was also examined. In mouse adipocytes, 1,25D repressed leptin mRNA levels by at least 84%, whereas 1,25D induced leptin mRNA 15.1-fold in human glioblastoma cells. Chromatin immunoprecipitation sequencing analysis of the mouse Lep gene in response to 1,25D revealed a cluster of regulatory sites (cis-regulatory module; CRM) at 228 kb that 1,25D-dependently docked VDR, RXR, C/EBPb, and RUNX2. This CRM harbored 3 VDREs and single C/EBPb and RUNX2 sites. Therefore, the expression of human TPH2 and mouse Lep are governed by 1,25D, potentially via respective VDREs located at 27/210 kb and 228 kb. These results imply that vitamin D affects brain serotonin concentrations, which may be relevant to psychiatric disorders, such as autism, and may control leptin levels and affect eating behavior.-Kaneko, I., Sabir, M. S., Dussik, C. M., Whitfield, G. K., Karrys, A., Hseih, J.-C., Haussler, M. R., Meyer, M. B., Pike, J. W., Jurutka, P. W. 1,25-dihydroxyvitamin D regulates expression of the tryptophan hydroxylase 2 and leptin genes: implication for behavioral influences of vitamin D. FASEB J. 29, 4023-4035 (2015 (1,25D), acts as a classic nuclear receptor ligand that binds specifically to the vitamin D receptor (VDR) to control the transcription of a multitude of genes, primarily those encoding proteins participating in bone mineral metabolism, regulating the immune system, and controlling cell growth and differentiation (1). Liganded VDR attracts one of the retinoid X receptors (RXRs) into a heterodimer that recognizes vitamin D responsive elements (VDREs) in the vicinity of target genes, regulating their expression via the recruitment of comodulator complexes that modify chromatin to effect either induction or repression of the cognate mRNA (2-4). A currently understudied area of vitamin D's function and significance is the CNS, even though the VDR protein is reportedly expressed broadly in the brain, including the neurons and glial cells (5). The most intense immunochemical signal is present in the hypothalamus (paraventricular and supraoptic nuclei and the lateral and ventromedial regions) and in the dopaminergic neurons of the substantia nigra (6). However, VDR protein is also detected in prefrontal cortex, cingulate gyrus, and CA2 region of the hippocampus. Thus, the VDR protein is clearly present in the CNS, but only limited information is available on the functions of VDR and its 1,25D ligand in the bra...