Palevitz B A
J Cell Biol. 1987 Jun;104(6):1515-9. doi: 10.1083/jcb.104.6.1515.
F-actin has been identified in the preprophase band of Allium cepa. Cells attached to subbed slides were obtained from formaldehyde-fixed root tips digested in EGTA and Cellulysin. The air-dried cells were extracted in Triton X-100, treated with rhodamine-phalloidin, rinsed briefly in PBS, and viewed in the fluorescence microscope. Interphase cells contain a network of actin fibers that extends into all areas of the cytoplasm. During preprophase, the network is replaced by a band of fibers aligned in the position of the preprophase band. Colocalization of F-actin with rhodamine-phalloidin and microtubules with tubulin immunocytochemistry confirms that the two bands are coincident. The actin appears to comprise a thin layer of fibers next to the plasmalemma. Like the microtubule preprophase band, the actin band narrows as preprophase progresses and disappears by midprophase. Fluorescent actin bands are not seen in fixed cells pretreated with excess unlabeled phalloidin before staining. They are also absent in roots exposed to cytochalasins B and D before fixation, but preprophase band microtubules at all stages of aggregation are still present. Colchicine treatment leads to the loss of both preprophase band microtubules and actin. The possible function of preprophase band actin is discussed.
已在洋葱的前期带中鉴定出F-肌动蛋白。附着在有涂层载玻片上的细胞取自用乙二醇双四乙酸(EGTA)和纤维素酶消化的甲醛固定根尖。将风干的细胞用曲拉通X-100提取,用罗丹明-鬼笔环肽处理,在磷酸盐缓冲盐水(PBS)中短暂冲洗,然后在荧光显微镜下观察。间期细胞含有延伸到细胞质所有区域的肌动蛋白纤维网络。在前期,该网络被一条纤维带取代,该纤维带排列在前期带的位置。F-肌动蛋白与罗丹明-鬼笔环肽的共定位以及微管与微管蛋白免疫细胞化学的共定位证实这两条带是重合的。肌动蛋白似乎在质膜旁边形成一层薄薄的纤维层。与微管前期带一样,随着前期的进展,肌动蛋白带变窄,并在中期消失。在用过量未标记的鬼笔环肽预处理后再染色的固定细胞中看不到荧光肌动蛋白带。在固定前暴露于细胞松弛素B和D的根中也不存在荧光肌动蛋白带,但在聚集的所有阶段前期带微管仍然存在。秋水仙碱处理导致前期带微管和肌动蛋白都消失。本文讨论了前期带肌动蛋白的可能功能。