Clark T G, Rosenbaum J L
Proc Natl Acad Sci U S A. 1982 Aug;79(15):4655-9. doi: 10.1073/pnas.79.15.4655.
Melanophore preparations of Fundulus heteroclitus that have been treated with the detergent Brij 58 can aggregate their pigment in response to epinephrine. On the basis of several criteria, it appears that cell lysis occurs under the detergent conditions used. Electron microscopic examination of detergent-treated cells shows progressive disruption of the melanophore plasma membrane during the time in which pigment aggregation occurs. Brij-treated cells are accessible to ferritin, a large electron-dense probe that is effectively excluded from non-detergent-treated controls. In cells incubated with detergent, fixed, and treated first with rat monoclonal antibodies against tubulin and then with fluorescein-isothiocyanate-labeled goat anti-rat IgG, a characteristic radical pattern of microtubule staining can be visualized by fluorescence microscopy. Control preparations treated similarly, but without detergent, do not stain. Vanadate, an inhibitor of ciliary and flagellar dynein ATPase, blocks melanosome aggregation in response to epinephrine in detergent-treated preparations but has no effect on intact melanophores. erythro-9-[3-(2-hydroxynonyl)]Adenine, another inhibitor of dynein ATPase, also inhibits pigment aggregation in Fundulus melanophores. The possibility that a dynein-like molecule plays a role in pigment aggregation is discussed.
用去污剂Brij 58处理过的底鳉黑素细胞制剂能在肾上腺素作用下聚集其色素。基于多项标准，在所用的去污剂条件下似乎发生了细胞裂解。对经去污剂处理的细胞进行电子显微镜检查显示，在色素聚集发生的时间段内，黑素细胞质膜逐渐受到破坏。Brij处理过的细胞可被铁蛋白（一种大的电子致密探针）进入，而该探针在未用去污剂处理的对照中被有效排除。在用去污剂孵育、固定并先用抗微管蛋白的大鼠单克隆抗体处理然后用异硫氰酸荧光素标记的山羊抗大鼠IgG处理的细胞中，通过荧光显微镜可观察到微管染色的特征性放射状模式。以类似方式处理但未用去污剂的对照制剂则不染色。钒酸盐是一种睫状和鞭毛动力蛋白ATP酶的抑制剂，它能阻断经去污剂处理的制剂中黑素体对肾上腺素的聚集反应，但对完整的黑素细胞没有影响。另一种动力蛋白ATP酶抑制剂erythro-9-[3-(2-羟基壬基)]腺嘌呤也能抑制底鳉黑素细胞中的色素聚集。文中讨论了一种类似动力蛋白的分子在色素聚集中发挥作用的可能性。
