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Aldosterone production is initiated by angiotensin II stimulation and activation of intracellular Ca²⁺ signaling. In aldosterone-producing adenoma (APA) cells, the activation of intracellular Ca²⁺ signaling is independent of the renin–angiotensin–aldosterone systems. The purpose of our study was to clarify molecular mechanisms of aldosterone production related to Ca²⁺ signaling. Transcriptome analysis revealed that the CALN1 gene encoding calneuron 1 had the strongest correlation with CYP11B2 (aldosterone synthase) among genes encoding Ca²⁺-binding proteins in APA. CALN1 modulation and synthetic or fluorescent compounds were used for functional studies in human adrenocortical carcinoma (HAC15) cells. CALN1 expression was 4.4-fold higher in APAs than nonfunctioning adrenocortical adenomas. CALN1 expression colocalized with CYP11B2 expression as investigated using …