Anoctamin-1 (ANO1; also termed TMEM16A) is a Ca²⁺-activated Cl^- (Cl_Ca) channel expressed in arterial myocytes that regulates membrane potential and contractility. Signaling mechanisms that control ANO1 activity in arterial myocytes are poorly understood. In cerebral artery myocytes, ANO1 channels are activated by local Ca²⁺ signals generated by plasma membrane non-selective cation channels, but the molecular identity of these proteins is unclear. Arterial myocytes express several different non-selective cation channels, including multiple members of the transient receptor potential receptor (TRP) family. The goal of this study was to identify localized ion channels that control ANO1 currents in cerebral artery myocytes. Co-immunoprecipitation and immunofluorescence resonance energy transfer microscopy experiments indicate that ANO1 and TRP canonical 6 (TRPC6) channels are present in the same macromolecular complex and locate in close spatial proximity in the myocyte plasma membrane. In contrast, ANO1 is not nearby TRPC3, TRPM4, or inositol trisphosphate receptor 1 (IP3R1) channels. Hyp9, a selective TRPC6 channel activator, stimulated Cl^- currents in myocytes that were blocked by T16Ainh-A01, an ANO1 inhibitor, ANO1 knockdown using siRNA, and equimolar replacement of intracellular EGTA with BAPTA, a fast Ca2+ chelator that abolishes local Ca²⁺ signaling. Hyp9 constricted pressurized cerebral arteries and this response was attenuated by T16Ainh-A01. In contrast T16Ainh-A01 did not alter depolarization-induced (60 mM K⁺) vasoconstriction. These data indicate that TRPC6 channels generate a local intracellular Ca²⁺ signal that activates nearby ANO1 channels in myocytes to stimulate vasoconstriction.