Aminochrome Induces Disruption of Actin, Alpha-, and Beta-Tubulin Cytoskeleton Networks in Substantia-Nigra-Derived Cell Line
Aminochrome Induces Disruption of Actin, Alpha-, and Beta-Tubulin Cytoskeleton Networks in Substantia-Nigra-Derived Cell Line
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DOI：
10.1007/s12640-009-9148-4
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发表时间：
2010-07-01
影响因子：
3.7
通讯作者：
Segura-Aguilar, Juan
中科院分区：
文献类型：
--
作者：
Paris, Irmgard;Perez-Pastene, Carolina;Segura-Aguilar, Juan
在以前的研究中，我们观察到，细胞处理的氨基色素通过氧化剂氧化多巴胺显着改变细胞形态，从而提出了这样的形态变化是否依赖于氨基色素或用于产生氨基色素的氧化剂的问题。因此，为了回答这个问题，我们现在已经在CM-Sepharose 50-100柱上纯化了氨基色素，并且使用NMR研究，我们已经证实所得氨基色素是纯的并且它保留了其结构。使用钙黄绿素-AM的荧光显微镜和透射电子显微镜显示，RCSN-3细胞呈细长形状，当细胞与50 μ M氨基色素或100 μ M双香豆醇（DT-心肌黄酶的抑制剂）孵育时，该形状没有变化。然而，当细胞在100 μ M双香豆素存在下与50 μ M氨基色素孵育时，细胞尺寸减小，细长形状变成球形。在这些条件下，肌动蛋白，α-和β-微管蛋白细胞骨架细丝网络成为凝聚在细胞膜周围。在连接培养细胞的细胞突起中也观察到肌动蛋白聚集体。这些结果表明，氨基色素单电子代谢诱导破坏的正常形态的肌动蛋白，α-和β-微管蛋白的细胞骨架，和DT-心肌黄酶防止这些影响。
In previous studies, we observed that cells treated with aminochrome obtained by oxidizing dopamine with oxidizing agents dramatically changed cell morphology, thus posing the question if such morphological changes were dependent on aminochrome or the oxidizing agents used to produce aminochrome. Therefore, to answer this question, we have now purified aminochrome on a CM-Sepharose 50-100 column and, using NMR studies, we have confirmed that the resulting aminochrome was pure and that it retained its structure. Fluorescence microscopy with calcein-AM and transmission electron microscopy showed that RCSN-3 cells presented an elongated shape that did not change when the cells were incubated with 50 mu M aminochrome or 100 mu M dicoumarol, an inhibitor of DT-diaphorase. However, the cell were reduced in size and the elongated shape become spherical when the cells where incubated with 50 mu M aminochrome in the presence of 100 mu M dicoumarol. Under these conditions, actin, alpha-, and beta-tubulin cytoskeleton filament networks became condensed around the cell membrane. Actin aggregates were also observed in cells processes that connected the cells in culture. These results suggest that aminochrome one-electron metabolism induces the disruption of the normal morphology of actin, alpha-, and beta-tubulin in the cytoskeleton, and that DT-diaphorase prevents these effects.