β-Catenin–dependent mechanotransduction dates back to the common ancestor of Cnidaria and Bilateria

Significance Besides genetic regulation, mechanical forces have been identified as important cues in numerous developmental processes. Mechanical forces can activate biochemical cascades in a process called mechanotransduction. Recent studies in vertebrates and flies elucidated the role of mechanical forces for mesodermal gene expression. However, it remains unclear whether mechanotransduction is a universal regulatory mechanism throughout Metazoa. Here, we show in the sea anemone Nematostella vectensis that mechanical pressure can ectopically activate or restore brachyury expression. This mechanotransduction is dependent on β-catenin, similar to vertebrates. We propose that a regulatory feedback loop between genetic and mechanical gene activation exists during gastrulation and the β-catenin–dependent mechanotransduction is an ancient regulatory mechanism, which was present in the common ancestor of cnidarians and bilaterians. Although the genetic regulation of cellular differentiation processes is well established, recent studies have revealed the role of mechanotransduction on a variety of biological processes, including regulation of gene expression. However, it remains unclear how universal and widespread mechanotransduction is in embryonic development of animals. Here, we investigate mechanosensitive gene expression during gastrulation of the starlet sea anemone Nematostella vectensis, a cnidarian model organism. We show that the blastoporal marker gene brachyury is down-regulated by blocking myosin II-dependent gastrulation movements. Brachyury expression can be restored by applying external mechanical force. Using CRISPR/Cas9 and morpholino antisense technology, we also show that mechanotransduction leading to brachyury expression is β-catenin dependent, similar to recent findings in fish and Drosophila [Brunet T, et al. (2013) Nat Commun 4:1–15]. Finally, we demonstrate that prolonged application of mechanical stress on the embryo leads to ectopic brachyury expression. Thus, our data indicate that β-catenin–dependent mechanotransduction is an ancient gene regulatory mechanism, which was present in the common ancestor of cnidarians and bilaterians, at least 600 million years ago.

[1]  J. Folkman,et al.  Role of cell shape in growth control , 1978, Nature.

[2]  H. Hidaka,et al.  Selective inhibition of catalytic activity of smooth muscle myosin light chain kinase. , 1987, The Journal of biological chemistry.

[3]  J. Smith,et al.  Expression of a xenopus homolog of Brachyury (T) is an immediate-early response to mesoderm induction , 1991, Cell.

[4]  D E Ingber,et al.  Mechanotransduction across the cell surface and through the cytoskeleton. , 1993, Science.

[5]  B. Herrmann,et al.  Homologs of the mouse Brachyury gene are involved in the specification of posterior terminal structures in Drosophila, Tribolium, and Locusta. , 1994, Genes & development.

[6]  C. S. Chen,et al.  Geometric control of cell life and death. , 1997, Science.

[7]  H. Bode,et al.  HyBra1, a Brachyury homologue, acts during head formation in Hydra. , 1999, Development.

[8]  H. Yamaguchi,et al.  Brachyury regulatory region active in embryonal carcinoma P19 cells. , 1999, Bioscience, biotechnology, and biochemistry.

[9]  A. Bauer,et al.  Brachyury is a target gene of the Wnt/β-catenin signaling pathway , 2000, Mechanisms of Development.

[10]  Peter G. Gillespie,et al.  Molecular basis of mechanosensory transduction , 2001, Nature.

[11]  D. Arendt,et al.  Evolution of the bilaterian larval foregut , 2001, Nature.

[12]  U. Technau.  Brachyury, the blastopore and the evolution of the mesoderm , 2001, BioEssays : news and reviews in molecular, cellular and developmental biology.

[13]  D. McClay,et al.  The role of Brachyury (T) during gastrulation movements in the sea urchin Lytechinus variegatus. , 2001, Developmental biology.

[14]  J. Spring,et al.  Conservation of Brachyury, Mef2, and Snail in the myogenic lineage of jellyfish: a connection to the mesoderm of bilateria. , 2002, Developmental biology.

[15]  B. Gumbiner,et al.  Zygotic Wnt activity is required for Brachyury expression in the early Xenopus laevis embryo. , 2002, Developmental biology.

[16]  U. Technau,et al.  Induction of gametogenesis in the basal cnidarian Nematostella vectensis (Anthozoa) , 2002, Development Genes and Evolution.

[17]  N. Lartillot,et al.  Expression pattern of Brachyury in the mollusc Patella vulgata suggests a conserved role in the establishment of the AP axis in Bilateria. , 2002, Development.

[18]  U. Technau,et al.  The ancestral role of Brachyury: expression of NemBra1 in the basal cnidarian Nematostella vectensis (Anthozoa) , 2002, Development Genes and Evolution.

[19]  P. Cohen,et al.  The specificities of protein kinase inhibitors: an update. , 2003, The Biochemical journal.

[20]  U. Technau,et al.  Arrested apoptosis of nurse cells during Hydra oogenesis and embryogenesis. , 2003, Developmental biology.

[21]  M. Martindale,et al.  An ancient role for nuclear β-catenin in the evolution of axial polarity and germ layer segregation , 2003, Nature.

[22]  E. Farge.  Mechanical Induction of Twist in the Drosophila Foregut/Stomodeal Primordium , 2003, Current Biology.

[23]  Gabriel Acevedo-Bolton,et al.  Intracardiac fluid forces are an essential epigenetic factor for embryonic cardiogenesis , 2003, Nature.

[24]  J. Smith,et al.  Evolution of Brachyury proteins: identification of a novel regulatory domain conserved within Bilateria. , 2003, Developmental biology.

[25]  Christopher S. Chen,et al.  Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. , 2004, Developmental cell.

[26]  U. Technau,et al.  Gastrulation in the sea anemone Nematostella vectensis occurs by invagination and immigration: an ultrastructural study , 2006, Development Genes and Evolution.

[27]  Alexander A Spector,et al.  Emergent patterns of growth controlled by multicellular form and mechanics. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[28]  M. Sheetz,et al.  Local force and geometry sensing regulate cell functions , 2006, Nature Reviews Molecular Cell Biology.

[29]  Lev V Beloussov,et al.  Gastrulation in amphibian embryos, regarded as a succession of biomechanical feedback events. , 2006, The International journal of developmental biology.

[30]  S. Sen,et al.  Matrix Elasticity Directs Stem Cell Lineage Specification , 2006, Cell.

[31]  M. Martindale,et al.  Surprisingly complex T‐box gene complement in diploblastic metazoans , 2007, Evolution & development.

[32]  M. Martindale,et al.  Gastrulation in the cnidarian Nematostella vectensis occurs via invagination not ingression. , 2007, Developmental biology.

[33]  Christopher S. Chen.  Mechanotransduction – a field pulling together? , 2008, Journal of Cell Science.

[34]  Lev V. Beloussov,et al.  Relocations of cell convergence sites and formation of pharyngula-like shapes in mechanically relaxed Xenopus embryos , 2008, Development Genes and Evolution.

[35]  Christopher S. Chen,et al.  Mechanotransduction in development: a growing role for contractility , 2009, Nature Reviews Molecular Cell Biology.

[36]  D. Ingber,et al.  Mechanotransduction at a distance: mechanically coupling the extracellular matrix with the nucleus , 2009, Nature Reviews Molecular Cell Biology.

[37]  Grigory Genikhovich,et al.  In situ hybridization of starlet sea anemone (Nematostella vectensis) embryos, larvae, and polyps. , 2009, Cold Spring Harbor protocols.

[38]  Grigory Genikhovich,et al.  Induction of spawning in the starlet sea anemone Nematostella vectensis, in vitro fertilization of gametes, and dejellying of zygotes. , 2009, Cold Spring Harbor protocols.

[39]  R. Harland,et al.  Endocytosis Is Required for Efficient Apical Constriction during Xenopus Gastrulation , 2010, Current Biology.

[40]  L V Beloussov,et al.  Neuro‐mesodermal patterns in artificially deformed embryonic explants: A role for mechano‐geometry in tissue differentiation , 2010, Developmental dynamics : an official publication of the American Association of Anatomists.

[41]  D. Ingber,et al.  Mechanochemical control of mesenchymal condensation and embryonic tooth organ formation. , 2011, Developmental cell.

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

[43]  Brenton D. Hoffman,et al.  Dynamic molecular processes mediate cellular mechanotransduction , 2011, Nature.

[44]  M. Martindale,et al.  A Framework for the Establishment of a Cnidarian Gene Regulatory Network for “Endomesoderm” Specification: The Inputs of ß-Catenin/TCF Signaling , 2012, PLoS genetics.

[45]  Y. Won,et al.  Estimation of divergence times in cnidarian evolution based on mitochondrial protein-coding genes and the fossil record. , 2012, Molecular phylogenetics and evolution.

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

[47]  J. Doudna,et al.  RNA-guided genetic silencing systems in bacteria and archaea , 2012, Nature.

[48]  T. Adachi,et al.  External mechanical cues trigger the establishment of the anterior-posterior axis in early mouse embryos. , 2013, Developmental cell.

[49]  Jeffry D. Sander,et al.  Efficient In Vivo Genome Editing Using RNA-Guided Nucleases , 2013, Nature Biotechnology.

[50]  W. Weis,et al.  The β-catenin destruction complex. , 2013, Cold Spring Harbor perspectives in biology.

[51]  Carl Paterson,et al.  Spectral broadening in Brillouin imaging , 2013 .

[52]  Frédéric Dumas-Bouchiat,et al.  Evolutionary conservation of early mesoderm specification by mechanotransduction in Bilateria , 2013, Nature Communications.

[53]  S. McKinney,et al.  TALEN and CRISPR/Cas9-mediated genome editing in the early-branching metazoan Nematostella vectensis , 2014, Nature Communications.

[54]  A. Martinez Arias,et al.  Brachyury cooperates with Wnt/β-catenin signalling to elicit primitive-streak-like behaviour in differentiating mouse embryonic stem cells , 2014, BMC Biology.

[55]  Michael P. Sheetz,et al.  Appreciating force and shape — the rise of mechanotransduction in cell biology , 2014, Nature Reviews Molecular Cell Biology.

[56]  Ziheng Yang,et al.  Uncertainty in the Timing of Origin of Animals and the Limits of Precision in Molecular Timescales , 2015, Current Biology.

[57]  William J. Polacheck,et al.  Noncontact three-dimensional mapping of intracellular hydro-mechanical properties by Brillouin microscopy , 2015, Nature Methods.

[58]  Grigory Genikhovich,et al.  Pre-bilaterian origin of the blastoporal axial organizer , 2016, Nature Communications.

[59]  Kareem Elsayad,et al.  Mapping the subcellular mechanical properties of live cells in tissues with fluorescence emission–Brillouin imaging , 2016, Science Signaling.

[60]  F. Rentzsch,et al.  Development of the aboral domain in Nematostella requires β-catenin and the opposing activities of Six3/6 and Frizzled5/8 , 2016, Development.

[61]  Giuseppe Antonacci,et al.  Biomechanics of subcellular structures by non-invasive Brillouin microscopy , 2016, Scientific Reports.

[62]  Patrick R. H. Steinmetz,et al.  Gut-like ectodermal tissue in a sea anemone challenges germ layer homology , 2017, Nature Ecology & Evolution.

[63]  Giuliano Scarcelli,et al.  Integration of spectral coronagraphy within VIPA-based spectrometers for high extinction Brillouin imaging. , 2017, Optics express.