Modulation of the subcellular distribution of calmodulin by melatonin in MDCK cells

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Abstract
In this paper we describe the interaction of melatonin with calmodulin in MDCK cells. The double staining immunofluorescent method showed that calmodulin in control MDCK cells appeared as fluorescent spots at the cell periphery. In contrast, MDCK cells cultured with 10(-9) M melatonin for 6, 12, 24 hr or for 4 days showed that calmodulin fluorescent spots were distributed throughout the cytoplasm as well as in the nucleus. These calmodulin rearrangements were reversed 6 hr after melatonin withdrawal. Moreover, calmodulin radioimmunoassay showed that in 10(-9) M melatonin-treated cells, membrane-bound calmodulin content was increased by 78% whereas cytosolic calmodulin decreased by 60%. Simultaneous labeling of MDCK cells with specific antibodies against calmodulin and melatonin showed that not only does the indole enter the cell, but that it has the same subcellular distribution as calmodulin. Besides the early responses induced by the melatonin antagonism to calmodulin, the results suggest that the indole may also induce long-term cellular responses by changing calmodulin concentrations in specific cellular compartments.