Histamine Regulates Molecular Clock Oscillations in Human Retinal Pigment Epithelial Cells via H₁ Receptors

Vertebrate eyes are known to contain circadian clocks, but their regulatory mechanisms remain largely unknown. To address this, we used a cell line from human retinal pigment epithelium (hRPE-YC) with stable coexpression of reporters for molecular clock oscillations (Bmal1-luciferase) and intracellular Ca²⁺ concentrations (YC3.6). We observed concentration-dependent increases in cytosolic Ca²⁺ concentrations after treatment with histamine (1-100 µM) and complete suppression of histamine-induced Ca²⁺ mobilizations by H₁ histamine receptor (H₁R) antagonist d-chlorpheniramine (d-CPA) in hRPE-YC cells. Consistently, real-time RT-PCR assays revealed that H₁R showed the highest expression among the four subtypes (H₁-H₄) of histamine receptors in hRPE-YC cells. Stimulation of hRPE-YC cells with histamine transiently increased nuclear localization of phosphorylated Ca²⁺/cAMP-response element-binding protein that regulates clock gene transcriptions. Administration of histamine also shifted the Bmal1-luciferase rhythms with a type-1 phase-response curve, similar to previous results with carbachol stimulations. Treatment of hRPE-YC cells with d-CPA or with more specific H₁R antagonist, ketotifen, blocked the histamine-induced phase shifts. Furthermore, an H₂ histamine receptor agonist, amthamine, had little effect on the Bmal1-luciferase rhythms. Although the function of the in vivo histaminergic system within the eye remains obscure, the present results suggest histaminergic control of the molecular clock via H₁R in retinal pigment epithelial cells. Also, since d-CPA and ketotifen have been widely used (e.g., to treat allergy and inflammation) in our daily life and thus raise a possible cause for circadian rhythm disorders by improper use of antihistamines.