Aims: The lipid-secreting exocrine Harderian gland contains a large amount of porphyrins (mainly protoporphyrin IX, PPIX) in the glandular cells, the physiological significance of which is rather poorly understood. Methods: In the present study, the possibility of using Fura-2 to measure intracellular calcium ([Ca2+]c) changes in these cells was assessed. Results: It was found that when Fura-2-loaded cells were excited by light at 340/380 nm, [Ca2+]c increased spontaneously, indicating a photodynamic action powered by light at 340/380 nm. In contrast, with the visible spectrum calcium probe Fluo-3 (λex = 475 nm), carbachol at 10 μM induced [Ca2+]c increase; [Ca 2+]c did not change without carbachol stimulation. Brief illumination with light at 340/380 nm induced a large [Ca2+] c increase in Fluo-3-loaded cells. Photodynamic stimulation of [Ca2+]c increase was confirmed with an exogenous photosensitizer sulphonated aluminium phthalocyanine (SALPC) and visible light (>580 nm). The wavelength-dependence of the [Ca2+]c increase correlates well with the excitation spectrum of the isolated Harderian glandular cells. Conclusion: These data suggest that PPIX present in rat Harderian glandular cells plays the role of a photosensitizer which upon activation by UVA and blue components of daylight and subsequent singlet oxygen generation, triggers [Ca2+]c increase and secretory response. The PPIX photodynamic action may also play a potential role in photic entrainment of the central circadian clock.
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