Regulators of cell movement during development and regeneration in Drosophila.

Cell migration is a fundamental cell biological process essential both for normal development and for tissue regeneration after damage. Cells can migrate individually or as a collective. To better understand the genetic requirements for collective migration, we expressed RNA interference (RNAi) against 30 genes in the Drosophila embryonic salivary gland cells that are known to migrate collectively.

Microtubule Motors in Establishment of Epithelial Cell Polarity.

Epithelial cells play a key role in insuring physiological homeostasis by acting as a barrier between the outside environment and internal organs. They are also responsible for the vectorial transport of ions and fluid essential to the function of many organs. To accomplish these tasks, epithelial cells must generate an asymmetrically organized plasma membrane comprised of structurally and functionally distinct apical and basolateral membranes.

Receptor-Type Guanylyl Cyclase at 76C (Gyc76C) Regulates De Novo Lumen Formation during Drosophila Tracheal Development.

Lumen formation and maintenance are important for the development and function of essential organs such as the lung, kidney and vasculature. In the Drosophila embryonic trachea, lumena form de novo to connect the different tracheal branches into an interconnected network of tubes. Here, we identify a novel role for the receptor type guanylyl cyclase at 76C (Gyc76C) in de novo lumen formation in the Drosophila trachea.

Receptor guanylyl cyclase Gyc76C is required for invagination, collective migration and lumen shape in the Drosophila embryonic salivary gland.

The Drosophila embryonic salivary gland is formed by the invagination and collective migration of cells. Here, we report on a novel developmental role for receptor-type guanylyl cyclase at 76C, Gyc76C, in morphogenesis of the salivary gland. We demonstrate that Gyc76C and downstream cGMP-dependent protein kinase 1 (DG1) function in the gland and surrounding mesoderm to control invagination, collective migration and lumen shape.

Rac1 GTPase acts downstream of αPS1βPS integrin to control collective migration and lumen size in the Drosophila salivary gland.

During collective migration of the Drosophila embryonic salivary gland, the distal gland cells mediate integrin-based contacts with surrounding tissues while proximal gland cells change shape and rearrange. Here, we show that αPS1βPS integrin controls salivary gland migration through Rac1 GTPase which downregulates E-cadherin in proximal and distal gland cells, and promotes extension of actin-rich basal membrane protrusions in the distal cells. In embryos mutant for multiple edematous wings (mew), which encodes the αPS1 subunit of the αPS1βPS integrin heterodimer, or rac1 and rac2 GTPases, salivary gland cells failed to migrate, to downregulate E-cadherin and to extend basal membrane protrusions.

Receptor-type guanylyl cyclase Gyc76C is required for development of the Drosophila embryonic somatic muscle.

Guanylyl cyclases mediate a number of physiological processes, including smooth muscle function and axonal guidance. Here, we report a novel role for Drosophila receptor-type guanylyl cyclase at 76C, Gyc76C, in development of the embryonic somatic muscle. In embryos lacking function of Gyc76C or the downstream cGMP-dependent protein kinase (cGK), DG1, patterning of the somatic body wall muscles was abnormal with ventral and lateral muscle groups showing the most severe defects.

Coordinated control of lumen size and collective migration in the salivary gland.

The Drosophila embryonic salivary gland is a powerful system to analyze the cellular and molecular mechanisms of tubulogenesis in vivo. The secretory portion of the salivary gland (referred to here as the salivary gland), is a pair of elongated epithelial tubes with a single central lumen, that is formed by the invagination and collective migration of gland cells from the ventral surface of the embryo. ( 1) (,) ( 2) As the salivary gland elongates during migration, the central lumen elongates and narrows at the proximal end.

Rho GTPase controls Drosophila salivary gland lumen size through regulation of the actin cytoskeleton and Moesin.

Generation and maintenance of proper lumen size is important for tubular organ function. We report on a novel role for the Drosophila Rho1 GTPase in control of salivary gland lumen size through regulation of cell rearrangement, apical domain elongation and cell shape change. We show that Rho1 controls cell rearrangement and apical domain elongation by promoting actin polymerization and regulating F-actin distribution at the apical and basolateral membranes through Rho kinase.

The bHLH transcription factor, hairy, refines the terminal cell fate in the Drosophila embryonic trachea.

Background: The pair-rule gene, hairy, encodes a basic helix-loop-helix transcription factor and is required for patterning of the early Drosophila embryo and for morphogenesis of the embryonic salivary gland. Although hairy was shown to be expressed in the tracheal primordia and in surrounding mesoderm, whether hairy plays a role in tracheal development is not known.

Principal Findings: Here, we report that hairy is required for refining the terminal cell fate in the embryonic trachea and that hairy's tracheal function is distinct from its earlier role in embryonic patterning.

Pak1 control of E-cadherin endocytosis regulates salivary gland lumen size and shape.

Generating and maintaining proper lumen size and shape in tubular organs is essential for organ function. Here, we demonstrate a novel role for p21-activated kinase 1 (Pak1) in defining the size and shape of the Drosophila embryonic salivary gland lumen by regulating the size and elongation of the apical domain of individual cells. Pak1 mediates these effects by decreasing and increasing E-cadherin levels at the adherens junctions and basolateral membrane, respectively, through Rab5- and Dynamin-dependent endocytosis.

Genetic regulation of salivary gland development in Drosophila melanogaster.

Studies of salivary gland development in the Drosophila embryo have revealed the morphogenic events by which the salivary gland tubes are formed, and identified the genes and genetic networks that regulate these events. Invagination of the salivary gland primordium occurs by an apical constriction mechanism regulated by the tyrosine kinase, Tec29, the transcription factors, Fork head and Huckebein, and Rho GTPase-mediated actomyosin contraction. After invagination is complete, transcriptional control of the apical membrane protein, Crumbs, by the transcription factors, Hairy, Hkb and Ribbon, and downregulation of Moesin-dependent apical stiffness promotes elongation of the salivary gland lumen.

Deficiency screen identifies a novel role for beta 2 tubulin in salivary gland and myoblast migration in the Drosophila embryo.

The Drosophila embryonic salivary gland is an epithelial organ formed by the coordinated invagination and migration of primordial cells. To identify genes that regulate gland migration we performed a deficiency screen of the third chromosome. Here, we report on the analysis of the beta 2 tubulin isoform (beta2t) that maps at 85D15.

Ribbon modulates apical membrane during tube elongation through Crumbs and Moesin.

Although the formation and maintenance of epithelial tubes are essential for the viability of multicellular organisms, our understanding of the molecular and cellular events coordinating tubulogenesis is relatively limited. Here, we focus on the activities of Ribbon, a novel BTB-domain containing nuclear protein, in the elongation of two epithelial tubes: the Drosophila salivary gland and trachea. We show that Ribbon interacts with Lola Like, another BTB-domain containing protein required for robust nuclear localization of Ribbon, to upregulate crumbs expression and downregulate Moesin activity.

Rho GTPase controls invagination and cohesive migration of the Drosophila salivary gland through Crumbs and Rho-kinase.

Coordinated cell movements shape simple epithelia into functional tissues and organs during embryogenesis. Regulators and effectors of the small GTPase Rho have been shown to be essential for epithelial morphogenesis in cell culture; however, the mechanism by which Rho GTPase and its downstream effectors control coordinated movement of epithelia in a developing tissue or organ is largely unknown. Here, we show that Rho1 GTPase activity is required for the invagination of Drosophila embryonic salivary gland epithelia and for directed migration of the internalized gland.

Rac function in epithelial tube morphogenesis.

Epithelial cell migration and morphogenesis require dynamic remodeling of the actin cytoskeleton and cell-cell adhesion complexes. Numerous studies in cell culture and in model organisms have demonstrated the small GTPase Rac to be a critical regulator of these processes; however, little is known about Rac function in the morphogenic movements that drive epithelial tube formation. Here, we use the embryonic salivary glands of Drosophila to understand the role of Rac in epithelial tube morphogenesis.

A molecular link between FGF and Dpp signaling in branch-specific migration of the Drosophila trachea.

The tracheal system of Drosophila embryos achieves its archetypal branching pattern through a series of cell migration events requiring the FGF, Dpp, and Wg/WNT signaling pathways. To gain insight into tracheal cell migration, we performed an F4 EMS mutagenesis screen to generate and characterize new mutations resulting in tracheal defects. From 2591 mutagenized third chromosome lines, we identified 33 mutations with defects in tracheal development, corresponding to 12 distinct complementation groups.

Making tubes in the Drosophila embryo.

Epithelial and endothelial tubes come in various shapes and sizes and form the basic units of many tubular organs. During embryonic development, single unbranched tubes as well as highly branched networks of tubes form from simple sheets of cells by several morphogenic movements. Studies of tube formation in the Drosophila embryo have greatly advanced our understanding of the cellular and molecular mechanisms by which tubes are formed.

Posterior migration of the salivary gland requires an intact visceral mesoderm and integrin function.

The final overall shape of an organ and its position within the developing embryo arise as a consequence of both its intrinsic properties and its interactions with surrounding tissues. Here, we focus on the role of directed cell migration in shaping and positioning the Drosophila salivary gland. We demonstrate that the salivary gland turns and migrates along the visceral mesoderm to become properly oriented with respect to the overall embryo.

Epithelial tube morphology is determined by the polarized growth and delivery of apical membrane.

Formation of tubes of the correct size and shape is essential for viability of most organisms, yet little is understood of the mechanisms controlling tube morphology. We identified a new allele of hairy in a mutagenesis screen and showed that hairy mutations cause branching and bulging of the normally unbranched salivary tube, in part through prolonged expression of huckebein (hkb). HKB controls polarized cell shape change and apical membrane growth during salivary cell invagination via two downstream target genes, crumbs (crb), a determinant of the apical membrane, and klarsicht (klar), which mediates microtubule-dependent organelle transport.