Kotz K J, McNiven M A
Department of Molecular Neuroscience, Mayo Clinic, Rochester, Minnesota 55905.
J Cell Biol. 1994 Feb;124(4):463-74. doi: 10.1083/jcb.124.4.463.
Teleost pigment cells (erythrophores and melanophores) are useful models for studying the regulation of rapid, microtubule-dependent organelle transport. Previous studies suggest that melanophores regulate the direction of pigment movements via changes in intracellular cAMP (Rozdzial and Haimo, 1986a; Sammak et al., 1992), whereas erythrophores may use calcium- (Ca(2+)-) based regulation (Luby-Phelps and Porter, 1982; McNiven and Ward, 1988). Despite these observations, there have been no direct measurements in intact erythrophores or any cell type correlating changes of intracellular free Ca2+ ([Ca2+]i) with organelle movements. Here we demonstrate that extracellular Ca2+ is necessary and that a Ca2+ influx via microinjection is sufficient to induce pigment aggregation in erythrophores, but not melanophores of squirrel fish. Using the Ca(2+)-sensitive indicator, Fura-2, we demonstrate that [Ca2+]i rises dramatically concomitant with aggregation of pigment granules in erythrophores, but not melanophores. In addition, we find that an erythrophore stimulated to aggregate pigment will immediately transmit a rise in [Ca2+]i to neighboring cells, suggesting that these cells are electrically coupled. Surprisingly, we find that a fall in [Ca2+]i is not sufficient to induce pigment dispersion in erythrophores, contrary to the findings obtained with the ionophore and lysed-cell models (Luby-Phelps and Porter, 1982; McNiven and Ward, 1988). We find that a rise in intracellular cAMP ([cAMP]i) induces pigment dispersion, and that this dispersive stimulus can be overridden by an aggregation stimulus, suggesting that both high [cAMP]i and low [Ca2+]i are necessary to produce pigment dispersion in erythrophores.
硬骨鱼色素细胞(红色素细胞和黑色素细胞)是研究快速的、微管依赖性细胞器运输调控的有用模型。先前的研究表明,黑色素细胞通过细胞内cAMP的变化来调节色素移动的方向(罗兹戴尔和海莫,1986a;萨马克等人,1992),而红色素细胞可能利用基于钙(Ca(2+))的调控(卢比-费尔普斯和波特,1982;麦克尼文和沃德,1988)。尽管有这些观察结果,但尚未在完整的红色素细胞或任何细胞类型中进行直接测量,以关联细胞内游离Ca2+([Ca2+]i)的变化与细胞器移动。在这里,我们证明细胞外Ca2+是必需的,并且通过显微注射的Ca2+内流足以诱导红色素细胞中的色素聚集,但对松鼠鱼的黑色素细胞则无效。使用Ca(2+)敏感指示剂Fura-2,我们证明红色素细胞中[Ca2+]i随着色素颗粒的聚集而急剧升高,但黑色素细胞中并非如此。此外,我们发现受到刺激而聚集色素的红色素细胞会立即将[Ca2+]i的升高传递给相邻细胞,这表明这些细胞是电耦合的。令人惊讶的是,我们发现与离子载体和裂解细胞模型的结果相反(卢比-费尔普斯和波特,1982;麦克尼文和沃德,1988),[Ca2+]i的下降不足以诱导红色素细胞中的色素分散。我们发现细胞内cAMP([cAMP]i)的升高会诱导色素分散,并且这种分散刺激可以被聚集刺激所抵消,这表明高[cAMP]i和低[Ca2+]i对于在红色素细胞中产生色素分散都是必需的。