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面包屑影响发育中的果蝇胚胎前区域的蛋白质动态。

Crumbs affects protein dynamics in anterior regions of the developing Drosophila embryo.

机构信息

Max-Planck-Institute of Molecular Cell Biology and Genetics, Dresden, Germany.

出版信息

PLoS One. 2013;8(3):e58839. doi: 10.1371/journal.pone.0058839. Epub 2013 Mar 21.

Abstract

Maintenance of apico-basal polarity is essential for epithelial integrity and requires particular reinforcement during tissue morphogenesis, when cells are reorganised, undergo shape changes and remodel their junctions. It is well established that epithelial integrity during morphogenetic processes depends on the dynamic exchange of adherens junction components, but our knowledge on the dynamics of other proteins and their dynamics during these processes is still limited. The early Drosophila embryo is an ideal system to study membrane dynamics during morphogenesis. Here, morphogenetic activities differ along the anterior-posterior axis, with the extending germband showing a high degree of epithelial remodelling. We developed a Fluorescence Recovery After Photobleaching (FRAP) assay with a higher temporal resolution, which allowed the distinction between a fast and a slow component of recovery of membrane proteins during the germband extension stage. We show for the first time that the recovery kinetics of a general membrane marker, SpiderGFP, differs in the anterior and posterior parts of the embryo, which correlates well with the different morphogenetic activities of the respective embryonic regions. Interestingly, absence of crumbs, a polarity regulator essential for epithelial integrity in the Drosophila embryo, decreases the fast component of SpiderGFP and of the apical marker Stranded at Second-Venus specifically in the anterior region. We suggest that the defects in kinetics observed in crumbs mutant embryos are the first signs of tissue instability in this region, explaining the earlier breakdown of the head epidermis in comparison to that of the trunk, and that diffusion in the plasma membrane is affected by the absence of Crumbs.

摘要

顶端-基底极性的维持对于上皮完整性至关重要,在组织形态发生过程中需要特别加强,此时细胞会被重新组织,发生形状变化并重塑它们的连接。上皮完整性在形态发生过程中依赖于黏着连接成分的动态交换,这一点已经得到充分证实,但是我们对其他蛋白质的动态及其在这些过程中的动力学的了解仍然有限。早期果蝇胚胎是研究形态发生过程中膜动态的理想系统。在这里,形态发生活动沿前后轴方向不同,延伸的原肠胚显示出高度的上皮重塑。我们开发了一种具有更高时间分辨率的荧光恢复后漂白(FRAP)测定法,可以区分原肠胚延伸阶段膜蛋白恢复的快和慢成分。我们首次表明,一般膜标记物 SpiderGFP 的恢复动力学在前部和后部胚胎之间存在差异,这与各自胚胎区域的不同形态发生活动很好地相关。有趣的是,缺失 crumbs,一种对于果蝇胚胎上皮完整性至关重要的极性调节剂,会特异性地降低前部区域中 SpiderGFP 和顶端标记物 Stranded at Second-Venus 的快成分。我们认为,crumbs 突变体胚胎中观察到的动力学缺陷是该区域组织不稳定的第一个迹象,这解释了与躯干相比头部表皮更早破裂的原因,并且质膜中的扩散受到 Crumbs 缺失的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f76/3605435/225331b9e924/pone.0058839.g001.jpg

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