Convergence and extension at gastrulation require a myosin IIB-dependent cortical actin network.
Force-producing convergence (narrowing) and extension (lengthening) of tissues by active intercalation of cells along the axis of convergence play a major role in axial morphogenesis during embryo development in both vertebrates and invertebrates, and failure of these processes in human embryos leads to defects including spina bifida and anencephaly. Here we use Xenopus laevis, a system in which the polarized cell motility that drives this active cell intercalation has been related to the development of forces that close the blastopore and elongate the body axis, to examine the role of myosin IIB in convergence and extension. We find that myosin IIB is localized in the cortex of intercalating cells, and show by morpholino knockdown that this myosin isoform is essential for the maintenance of a stereotypical, cortical actin cytoskeleton as visualized with time-lapse fluorescent confocal microscopy. We show that this actin network consists of foci or nodes connected by cables and is polarized relative to the embryonic axis, preferentially cyclically shortening and lengthening parallel to the axis of cell polarization, elongation and intercalation, and also parallel to the axis of convergence forces during gastrulation. Depletion of MHC-B results in disruption of this polarized cytoskeleton, loss of the polarized protrusive activity characteristic of intercalating cells, eventual loss of cell-cell and cell-matrix adhesion, and dose-dependent failure of blastopore closure, arguably because of failure to develop convergence forces parallel to the myosin IIB-dependent dynamics of the actin cytoskeleton. These findings bridge the gap between a molecular-scale motor protein and tissue-scale embryonic morphogenesis.
Department of Biology, University of Virginia, Charlottesville, VA 22903, USA. frogman@virginia.edu
Paul
P
Skoglund
Ana
A
Rolo
Xuejun
X
Chen
Barry M
BM
Gumbiner
Ray
R
Keller
D000199:Actins / Q000502:physiology*; D000818:Animals; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion; D002465:Cell Movement; D003599:Cytoskeleton / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000502:physiology; D005260:Female; D005775:Gastrula / Q000187:drug effects; D054262:Gastrulation / Q000502:physiology*; D039482:Integrin alpha5 / Q000378:metabolism; D008565:Membrane Proteins / Q000502:physiology; D009024:Morphogenesis; D024501:Nonmuscle Myosin Type IIB / Q000378:metabolism / Q000502:physiology*; D009672:Notochord / Q000166:cytology / Q000502:physiology; D020033:Protein Isoforms / Q000378:metabolism / Q000502:physiology; D014981:Xenopus / Q000196:embryology; D029867:Xenopus Proteins / Q000378:metabolism
D000199:Actins; C074260:CDH3 protein, Xenopus; D015820:Cadherins; D039482:Integrin alpha5; D008565:Membrane Proteins; D020033:Protein Isoforms; D029867:Xenopus Proteins; D024501:Nonmuscle Myosin Type IIB