An Optogenetic Method to Modulate Cell Contractility during Tissue Morphogenesis

Summary Morphogenesis of multicellular organisms is driven by localized cell shape changes. How, and to what extent, changes in behavior in single cells or groups of cells influence neighboring cells and large-scale tissue remodeling remains an open question. Indeed, our understanding of multicellular dynamics is limited by the lack of methods allowing the modulation of cell behavior with high spatiotemporal precision. Here, we developed an optogenetic approach to achieve local modulation of cell contractility and used it to control morphogenetic movements during Drosophila embryogenesis. We show that local inhibition of apical constriction is sufficient to cause a global arrest of mesoderm invagination. By varying the spatial pattern of inhibition during invagination, we further demonstrate that coordinated contractile behavior responds to local tissue geometrical constraints. Together, these results show the efficacy of this optogenetic approach to dissect the interplay between cell-cell interaction, force transmission, and tissue geometry during complex morphogenetic processes.

[1]  M. Peifer,et al.  Abelson kinase (Abl) and RhoGEF2 regulate actin organization during cell constriction in Drosophila , 2006, Development.

[2]  Carsten Schultz,et al.  Protein translocation as a tool: The current rapamycin story , 2012, FEBS letters.

[3]  S. Breuer,et al.  skittles, a Drosophila phosphatidylinositol 4-phosphate 5-kinase, is required for cell viability, germline development and bristle morphology, but not for neurotransmitter release. , 1998, Genetics.

[4]  S. Roth,et al.  Autonomy and non-autonomy in Drosophila mesoderm determination and morphogenesis. , 1994, Development.

[5]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[6]  M. Miodownik,et al.  Robust mechanisms of ventral furrow invagination require the combination of cellular shape changes , 2009, Physical biology.

[7]  Bob Goldstein,et al.  Apical constriction: themes and variations on a cellular mechanism driving morphogenesis , 2014, Development.

[8]  M. Peifer,et al.  How the cytoskeleton helps build the embryonic body plan: models of morphogenesis from Drosophila. , 2009, Current topics in developmental biology.

[9]  C. Parent,et al.  Phosphoinositides Specify Polarity during Epithelial Organ Development , 2007, Cell.

[10]  T. Lecuit,et al.  Trafficking through Rab11 Endosomes Is Required for Cellularization during Drosophila Embryogenesis , 2003, Current Biology.

[11]  Christopher A. Voigt,et al.  Spatiotemporal Control of Cell Signalling Using A Light-Switchable Protein Interaction , 2009, Nature.

[12]  Y. Bellaïche,et al.  PTEN controls junction lengthening and stability during cell rearrangement in epithelial tissue. , 2013, Developmental cell.

[13]  Pierre-François Lenne,et al.  Nature and anisotropy of cortical forces orienting Drosophila tissue morphogenesis , 2008, Nature Cell Biology.

[14]  Adam C. Martin,et al.  Apical domain polarization localizes actin-myosin activity to drive ratchet-like apical constriction , 2013, Nature Cell Biology.

[15]  Felix Oswald,et al.  Forces Driving Epithelial Spreading in Zebrafish Gastrulation , 2012, Science.

[16]  Jared E. Toettcher,et al.  Using Optogenetics to Interrogate the Dynamic Control of Signal Transmission by the Ras/Erk Module , 2013, Cell.

[17]  E. Wieschaus,et al.  Gastrulation in Drosophila: the formation of the ventral furrow and posterior midgut invaginations. , 1991, Development.

[18]  Maria Leptin,et al.  Control of Drosophila Gastrulation by Apical Localization of Adherens Junctions and RhoGEF2 , 2007, Science.

[19]  J. Lessard,et al.  Actin distribution patterns in the mouse neural tube during neurulation. , 1982, Science.

[20]  A. Miyawaki,et al.  Two-photon dual-color imaging using fluorescent proteins , 2008, Nature Methods.

[21]  M. Miodownik,et al.  A Biomechanical Analysis of Ventral Furrow Formation in the Drosophila Melanogaster Embryo , 2012, PloS one.

[22]  K D Irvine,et al.  Cell intercalation during Drosophila germband extension and its regulation by pair-rule segmentation genes. , 1994, Development.

[23]  Pierre-François Lenne,et al.  Force generation, transmission, and integration during cell and tissue morphogenesis. , 2011, Annual review of cell and developmental biology.

[24]  S. Colombo,et al.  PI(4,5)P2-Dependent and Ca2+-Regulated ER-PM Interactions Mediated by the Extended Synaptotagmins , 2013, Cell.

[25]  M. Roth,et al.  Phosphatidylinositol 4,5-bisphosphate induces actin-based movement of raft-enriched vesicles through WASP-Arp2/3 , 2000, Current Biology.

[26]  Christopher A. Voigt,et al.  The promise of optogenetics in cell biology: interrogating molecular circuits in space and time , 2011, Nature Methods.

[27]  Jennifer A Zallen,et al.  Patterned gene expression directs bipolar planar polarity in Drosophila. , 2004, Developmental cell.

[28]  Wolfgang Huber,et al.  EBImage—an R package for image processing with applications to cellular phenotypes , 2010, Bioinform..

[29]  C. Heisenberg,et al.  Forces in Tissue Morphogenesis and Patterning , 2013, Cell.

[30]  P. Majerus,et al.  The protein deficient in Lowe syndrome is a phosphatidylinositol-4,5-bisphosphate 5-phosphatase. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[31]  P. De Camilli,et al.  Optogenetic control of phosphoinositide metabolism , 2012, Proceedings of the National Academy of Sciences.

[32]  C. Schultz,et al.  Plasma membrane phosphoinositide balance regulates cell shape during Drosophila embryo morphogenesis , 2014, The Journal of cell biology.

[33]  E. Munro,et al.  Sequential Activation of Apical and Basolateral Contractility Drives Ascidian Endoderm Invagination , 2010, Current Biology.

[34]  S. Parkhurst,et al.  Cell wound repair in Drosophila occurs through three distinct phases of membrane and cytoskeletal remodeling , 2011, The Journal of cell biology.

[35]  P. Devreotes,et al.  Phosphoinositide signaling plays a key role in cytokinesis , 2006, The Journal of cell biology.

[36]  Julien Colombelli,et al.  Pulsed Forces Timed by a Ratchet-like Mechanism Drive Directed Tissue Movement during Dorsal Closure , 2009, Cell.

[37]  L. Sulak,et al.  Myosin-dependent junction remodelling controls planar cell intercalation and axis elongation , 2004, Nature.

[38]  Chentao Lin,et al.  Photoexcited CRY2 Interacts with CIB1 to Regulate Transcription and Floral Initiation in Arabidopsis , 2008, Science.

[39]  Ravi S Kane,et al.  Optogenetic protein clustering and signaling activation in mammalian cells , 2013, Nature Methods.

[40]  Eric F. Wieschaus,et al.  Integration of contractile forces during tissue invagination , 2010, The Journal of cell biology.

[41]  Willy Supatto,et al.  Dynamic Analyses of Drosophila Gastrulation Provide Insights into Collective Cell Migration , 2008, Science.

[42]  Adam C. Martin,et al.  Intracellular signalling and intercellular coupling coordinate heterogeneous contractile events to facilitate tissue folding , 2015, Nature Communications.

[43]  W. Denk,et al.  Two-photon laser scanning fluorescence microscopy. , 1990, Science.

[44]  J. Settleman,et al.  The Rho GTPase and a Putative RhoGEF Mediate a Signaling Pathway for the Cell Shape Changes in Drosophila Gastrulation , 1997, Cell.

[45]  S. Grill,et al.  Anisotropies in cortical tension reveal the physical basis of polarizing cortical flows , 2010, Nature.

[46]  Eric F. Wieschaus,et al.  Pulsed contractions of an actin–myosin network drive apical constriction , 2009, Nature.