Inhibition of SOCE disrupts cytokinesis in zebrafish embryos via inhibition of cleavage furrow deepening.

C. Chan, Yiyun Chen, T. S. Hung, A. Miller, A. Shipley, S. Webb
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引用次数: 16

Abstract

During the first few cell division cycles in zebrafish, distinct Ca(2+) transients are localized to the early embryonic cleavage furrows, where they accompany (and are required for) furrow positioning, propagation, deepening and apposition. It has previously been shown that the endoplasmic reticulum (ER) acts as the primary store for generating these Ca(2+) transients via release through inositol 1,4,5-trisphosphate receptors (IP 3Rs). We hypothesised that maintaining the elevated levels of intracellular Ca(2+) required for deepening and apposition of the cleavage furrows in these large eggs might result in the depletion of the available ER Ca(2+) store, thus the role of store-operated Ca(2+) entry (SOCE) was examined. Newly fertilized, dechorionated embryos were incubated with various SOCE inhibitors, starting just prior to the onset of the first cell division cycle. The effect of these inhibitors on mitosis, furrow positioning, propagation, deepening and apposition, and the generation of the cytokinetic Ca(2+) transients was determined. Treatment with 2-APB or SKF 96365 had no major effect on mitosis, furrow positioning or propagation, but inhibited furrow deepening resulting in regression of the cleavage furrow. Both of these inhibitors also blocked the furrowing Ca(2+) transient, with SKF 96365 having a more profound inhibitory effect than 2-APB. In zebrafish, SOCE does not appear to be required for mitosis or the early stages of cytokinesis during the early embryonic cell division cycles, but it does appear to be essential for maintaining the elevated levels of [Ca(2+)]i for the extended periods that are required during furrow deepening and daughter cell apposition.
抑制SOCE通过抑制卵裂沟加深破坏斑马鱼胚胎的细胞分裂。
在斑马鱼最初的几个细胞分裂周期中,不同的Ca(2+)瞬态定位于早期胚胎分裂沟,在那里它们伴随着(并且是必需的)沟的定位、繁殖、加深和合并。先前的研究表明,内质网(ER)通过肌醇1,4,5-三磷酸受体(IP 3Rs)释放,作为产生这些Ca(2+)瞬态的主要储存。我们假设,在这些大卵中,维持细胞内钙(2+)的升高水平,以加深和放置卵裂沟,可能导致可用ER钙(2+)储存的耗尽,因此,我们研究了储存操作的钙(2+)进入(SOCE)的作用。在第一个细胞分裂周期开始之前,用各种SOCE抑制剂孵育新受精、脱去绒毛膜的胚胎。测定了这些抑制剂对有丝分裂、沟定位、繁殖、加深和附着以及细胞动力学Ca(2+)瞬态的产生的影响。2-APB或SKF 96365处理对有丝分裂、沟定位和繁殖没有明显影响,但抑制了沟加深,导致卵裂沟消退。这两种抑制剂也阻断了沟状Ca(2+)瞬态,其中SKF 96365比2- apb具有更深刻的抑制作用。在斑马鱼中,在早期胚胎细胞分裂周期中,有丝分裂或细胞质分裂的早期阶段似乎并不需要SOCE,但在沟槽加深和子细胞增生所需的较长时间内,SOCE对于维持[Ca(2+)]i的升高水平似乎是必不可少的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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