Melatonin-induced desensitization in amphibian melanophores

Video-microscopic examination of pigment granule translocation in cultured amphibian melanophores provides continuous, real-time observation of cellular responses to hormonal and pharmacologic agents and is particularly useful for studying the mechanisms underlying melatonin-induced pigment aggregation. We have used such video-microscopic technology to show that pigment cells become refractory to prolonged melatonin treatment and that the speed at which the desensitized condition becomes evident is dependent upon the countervailing concentration of antagonistic hormone, alpha-melanocyte stimulating hormone (MSH). When melanophores were treated with 10 nM melatonin, desensitization occurred within 30 minutes in the presence of 5 ng/ml MSH, whereas five hours of melatonin treatment was required before the desensitized state was observed in the presence of 1 ng/ml MSH. The persistence of desensitization after melatonin removal depends upon the duration of initial melatonin exposure. When melanophores were treated with melatonin (10 nM) in the presence of 10 ng/ml MSH for two hours, the desensitized condition lasted less than 30 minutes; if the initial melatonin treatment was increased to four hours, however, the melanophores remained in the desensitized state for more than two hours after the melatonin was removed from the medium. During the course of these treatments, there was no substantial degradation of melatonin activity; i.e., a second population of melanophores responded normally to the melatonin-containing media overlying desensitized melanophores.