Borges Ricardo Moraes, Lamers Marcelo Lazzaron, Forti Fabio Luis, Santos Marinilce Fagundes Dos, Yan Chao Yun Irene
Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, SP, Brazil.
Genesis. 2011 May;49(5):368-79. doi: 10.1002/dvg.20723. Epub 2011 Apr 2.
Epithelial invagination in many model systems is driven by apical cell constriction, mediated by actin and myosin II contraction regulated by GTPase activity. Here we investigate apical constriction during chick lens placode invagination. Inhibition of actin polymerization and myosin II activity by cytochalasin D or blebbistatin prevents lens invagination. To further verify if lens placode invaginate through apical constriction, we analyzed the role of Rho-ROCK pathway. Rho GTPases expression at the apical portion of the lens placode occurs with the same dynamics as that of the cytoskeleton. Overexpression of the pan-Rho inhibitor C3 exotoxin abolished invagination and had a strong effect on apical myosin II enrichment and a mild effect on apical actin localization. In contrast, pharmacological inhibition of ROCK activity interfered significantly with apical enrichment of both actin and myosin. These results suggest that apical constriction in lens invagination involves ROCK but apical concentration of actin and myosin are regulated through different pathways upstream of ROCK. genesis 49:368-379, 2011.
在许多模型系统中,上皮内陷是由顶端细胞收缩驱动的,这种收缩由受GTPase活性调节的肌动蛋白和肌球蛋白II介导。在此,我们研究鸡晶状体原基内陷过程中的顶端收缩。用细胞松弛素D或blebbistatin抑制肌动蛋白聚合和肌球蛋白II活性可阻止晶状体内陷。为进一步验证晶状体原基是否通过顶端收缩内陷,我们分析了Rho-ROCK途径的作用。晶状体原基顶端部分的Rho GTPases表达与细胞骨架的表达具有相同的动态变化。泛Rho抑制剂C3外毒素的过表达消除了内陷,并对顶端肌球蛋白II的富集有强烈影响,对顶端肌动蛋白定位有轻微影响。相反,ROCK活性的药理学抑制显著干扰了肌动蛋白和肌球蛋白的顶端富集。这些结果表明,晶状体内陷中的顶端收缩涉及ROCK,但肌动蛋白和肌球蛋白的顶端聚集是通过ROCK上游的不同途径调节的。《发生学》49:368 - 379,2011年。