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系统表达和功能丧失分析定义了果蝇 RhoGEFs 和 RhoGAPs 在腿部形态发生中空间限制的要求。

Systematic expression and loss-of-function analysis defines spatially restricted requirements for Drosophila RhoGEFs and RhoGAPs in leg morphogenesis.

机构信息

Department of Anatomy and Cellular Biology, Program in Cell, Molecular and Developmental Biology, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA 02111, USA.

出版信息

Mech Dev. 2011 Jan-Feb;128(1-2):5-17. doi: 10.1016/j.mod.2010.09.001. Epub 2010 Sep 17.

Abstract

The Drosophila leg imaginal disc consists of a peripheral region that contributes to adult body wall, and a central region that forms the leg proper. While the patterning signals and transcription factors that determine the identity of adult structures have been identified, the mechanisms that determine the shape of these structures remain largely unknown. The family of Rho GTPases, which consists of seven members in flies, modulates cell adhesion, actomyosin contractility, protrusive membrane activity, and cell-matrix adhesion to generate mechanical forces that shape adult structures. The Rho GTPases are ubiquitously expressed and it remains unclear how they orchestrate morphogenetic events. The Rho guanine nucleotide exchange factors (RhoGEFs) and Rho GTPase activating proteins (RhoGAPs), which respectively activate and deactivate corresponding Rho GTPases, have been proposed to regulate the activity of Rho signaling cascades in specific spatiotemporal patterns to orchestrate morphogenetic events. Here we identify restricted expression of 12 of the 20 RhoGEFs and 10 of the 22 Rho RhoGAPs encoded in Drosophila during metamorphosis. Expression of a subset of each family of RhoGTPase regulators was restricted to motile cell populations including tendon, muscle, trachea, and peripodial stalk cells. A second subset was restricted either to all presumptive joints or only to presumptive tarsal joints. Depletion of individual RhoGEFs and RhoGAPs in the epithelium of the disc proper identified several joint-specific genes, which act downstream of segmental patterning signals to control epithelial morphogenesis. Our studies provide a framework with which to understand how Rho signaling cascades orchestrate complex morphogenetic events in multi-cellular organisms, and evidence that patterning signals regulate these cascades to control apical constriction and epithelial invagination at presumptive joints.

摘要

果蝇腿的翅芽盘由一个周边区域组成,该区域有助于形成成虫体壁,以及一个中央区域,该区域形成腿的主体。虽然确定了决定成虫结构身份的模式信号和转录因子,但决定这些结构形状的机制在很大程度上仍然未知。Rho GTPase 家族由果蝇中的 7 个成员组成,调节细胞黏附、肌动球蛋白收缩性、突出膜活性和细胞-基质黏附,以产生塑造成虫结构的机械力。Rho GTPases 广泛表达,其如何协调形态发生事件仍不清楚。Rho 鸟嘌呤核苷酸交换因子(RhoGEFs)和 Rho GTPase 激活蛋白(RhoGAPs)分别激活和失活相应的 Rho GTPases,据推测,它们以特定的时空模式调节 Rho 信号级联的活性,以协调形态发生事件。在这里,我们在果蝇的变态过程中鉴定了 20 个 RhoGEFs 中的 12 个和 22 个 RhoGAPs 中的 10 个的受限表达。这些 RhoGTPase 调节剂家族的一部分表达仅限于包括腱、肌肉、气管和足垫柄细胞在内的运动细胞群体。另一部分子集仅限于所有假定的关节或仅针对假定的跗关节。在盘状上皮细胞中耗尽单个 RhoGEFs 和 RhoGAPs,鉴定出几个关节特异性基因,这些基因作为节段性模式信号的下游因子,控制上皮形态发生。我们的研究提供了一个框架,以了解 Rho 信号级联如何协调多细胞生物中的复杂形态发生事件,并提供证据表明模式信号调节这些级联以控制假定关节处的顶端缢缩和上皮内陷。

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