Brain-derived neurotrophic factor (BDNF) is an important mediator of activity-dependent functions of the nervous system and its expression is dysregulated in several neuropsychiatric disorders. Regulation of rodentBDNFneuronal activity-dependent transcription has been relatively well characterized. Here, we have studied regulation of humanBDNF(hBDNF) transcription by membrane depolarization of cultured mouse or rat primary cortical neurons expressing hBDNFgene or transfected with hBDNFpromoter constructs, respectively. We identified an asymmetric E-box-like element, PasRE [basic helix-loop-helix (bHLH)-PAS transcription factor response element], in hBDNFpromoter I and demonstrate that binding of this element by bHLH-PAS transcription factors ARNT2 (aryl hydrocarbon receptor nuclear translocator 2) and NPAS4 (neuronal PAS domain protein 4) is crucial for neuronal activity-dependent transcription from promoter I. We show that binding of CREB (cAMP response element-binding protein) to the cAMP/Ca²⁺-response element (CRE) in hBDNFpromoter IV is critical for activity-dependent transcription from this promoter and that upstream stimulatory factor (USF) transcription factors also contribute to the activation by binding to the upstream stimulatory factor binding element (UBE) in hBDNFpromoter IV. However, we report that full induction of hBDNFexon IV mRNA transcription is dependent on ARNT2 and NPAS4 binding to a PasRE in promoter IV. Finally, we demonstrate that CRE and PasRE elements in hBDNFpromoter IX are required for the induction of this promoter by neuronal activity. Together, the results of this study have identified thecis-elements and transcription factors regulating neuronal activity-dependent transcription of humanBDNFgene.