Interactions between mural cells and endothelial cells stabilize the developing zebrafish dorsal aorta

Mural cells (vascular smooth muscle cells and pericytes) play an essential role in the development of the vasculature, promoting vascular quiescence and long-term vessel stabilization through their interactions with endothelial cells. However, the mechanistic details of how mural cells stabilize vessels are not fully understood. We have examined the emergence and functional role of mural cells investing the dorsal aorta during early development using the zebrafish. Consistent with previous literature, our data suggest that cells ensheathing the dorsal aorta emerge from a sub-population of cells in the adjacent sclerotome. Inhibition of mural cell recruitment to the dorsal aorta through disruption of pdgfr signaling leads to a reduced vascular basement membrane, which in turn results in enhanced dorsal aorta vessel elasticity and failure to restrict aortic diameter. Our results provide direct in vivo evidence for a functional role for mural cells in patterning and stabilization of the early vasculature through production and maintenance of the vascular basement membrane to prevent abnormal aortic expansion and elasticity. Summary: Vascular smooth muscle cells are required to help maintain the vascular basement membrane and promote vessel stabilization during development.

[1]  J. Kitajewski,et al.  Combined deficiency of Notch1 and Notch3 causes pericyte dysfunction, models CADASIL, and results in arteriovenous malformations , 2015, Scientific Reports.

[2]  S. Weiss,et al.  An MT1-MMP-PDGF receptor-beta axis regulates mural cell investment of the microvasculature. , 2005, Genes & development.

[3]  E. Knapik,et al.  An exclusively mesodermal origin of fin mesenchyme demonstrates that zebrafish trunk neural crest does not generate ectomesenchyme , 2013, Development.

[4]  Amber N. Stratman,et al.  Endothelial-derived PDGF-BB and HB-EGF coordinately regulate pericyte recruitment during vasculogenic tube assembly and stabilization. , 2010, Blood.

[5]  Bengt R. Johansson,et al.  Pericytes regulate the blood–brain barrier , 2010, Nature.

[6]  P. Yurchenco.  Basement membranes: cell scaffoldings and signaling platforms. , 2011, Cold Spring Harbor perspectives in biology.

[7]  Ruijin Huang,et al.  Sclerotomal origin of smooth muscle cells in the wall of the avian dorsal aorta , 2007, Developmental dynamics : an official publication of the American Association of Anatomists.

[8]  C. Lien,et al.  Platelet-Derived Growth Factor Receptor β Is Critical for Zebrafish Intersegmental Vessel Formation , 2010, PloS one.

[9]  D. Lu,et al.  Molecular cloning and expression of a smooth muscle-specific gene SM22α in zebrafish☆ , 2003 .

[10]  A. Kudo,et al.  Twist functions in vertebral column formation in medaka, Oryzias latipes , 2004, Mechanisms of Development.

[11]  R. Kelsh,et al.  Clarification of mural cell coverage of vascular endothelial cells by live imaging of zebrafish , 2016, Development.

[12]  E. Keshet,et al.  A plasticity window for blood vessel remodelling is defined by pericyte coverage of the preformed endothelial network and is regulated by PDGF-B and VEGF. , 1998, Development.

[13]  J. Abrams,et al.  Characterization of zebrafish intestinal smooth muscle development using a novel sm22α‐b promoter , 2010, Developmental dynamics : an official publication of the American Association of Anatomists.

[14]  A. Aplin,et al.  MCP-1 promotes mural cell recruitment during angiogenesis in the aortic ring model , 2010, Angiogenesis.

[15]  B R Johansson,et al.  Pericyte loss and microaneurysm formation in PDGF-B-deficient mice. , 1997, Science.

[16]  W. Burggren,et al.  A Three‐Dimensional Functional Assessment of Heart and Vessel Development in the Larva of the Zebrafish (Danio rerio) , 2005, Physiological and Biochemical Zoology.

[17]  G. Davis,et al.  Biosynthesis, Remodeling, and Functions During Vascular Morphogenesis and Neovessel Stabilization , 2005 .

[18]  M. Majesky.  Developmental basis of vascular smooth muscle diversity. , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[19]  D. Davidson,et al.  Central roles of alpha5beta1 integrin and fibronectin in vascular development in mouse embryos and embryoid bodies. , 2002, Arteriosclerosis, thrombosis, and vascular biology.

[20]  W. Talbot,et al.  notch3 is essential for oligodendrocyte development and vascular integrity in zebrafish , 2013, Disease Models & Mechanisms.

[21]  B. Venkatesh,et al.  Phylogenetic and evolutionary relationships and developmental expression patterns of the zebrafish twist gene family , 2009, Development Genes and Evolution.

[22]  A. Eszterhas,et al.  TGF-β1 modulates EGF-stimulated phosphatidylinositol 3-kinase activity in human airway smooth muscle cells. , 1997, American journal of physiology. Lung cellular and molecular physiology.

[23]  K. Alitalo,et al.  Angiopoietin‐regulated recruitment of vascular smooth muscle cells by endothelial‐derived heparin binding EGF‐like growth factor , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[24]  M. Klagsbrun,et al.  Heparin-binding EGF-like growth factor stimulation of smooth muscle cell migration: dependence on interactions with cell surface heparan sulfate , 1993, The Journal of cell biology.

[25]  G. Owens,et al.  Regulation of differentiation of vascular smooth muscle cells. , 1995, Physiological reviews.

[26]  U. Landegren,et al.  Endothelial PDGF-B retention is required for proper investment of pericytes in the microvessel wall. , 2003, Genes & development.

[27]  C. Betsholtz,et al.  Pericytes: developmental, physiological, and pathological perspectives, problems, and promises. , 2011, Developmental cell.

[28]  David J. Anderson,et al.  Ventromedial hypothalamic neurons control a defensive emotion state , 2015, eLife.

[29]  N. Fusenig,et al.  Dynamics of basement membrane formation by keratinocyte-fibroblast interactions in organotypic skin culture. , 1998, Experimental cell research.

[30]  Kendal Broadie,et al.  CNS-derived glia ensheath peripheral nerves and mediate motor root development , 2008, Nature Neuroscience.

[31]  J. Nathans,et al.  Tip cell-specific requirement for an atypical Gpr124- and Reck-dependent Wnt/β-catenin pathway during brain angiogenesis , 2015, eLife.

[32]  R. Hynes,et al.  Defects in mesoderm, neural tube and vascular development in mouse embryos lacking fibronectin. , 1993, Development.

[33]  K. Hirschi,et al.  Endothelial cells modulate the proliferation of mural cell precursors via platelet-derived growth factor-BB and heterotypic cell contact. , 1999, Circulation research.

[34]  T. Ji,et al.  Cross-linking of fibronectin to sulfated proteoglycans at the cell surface , 1979, Cell.

[35]  Steven Song,et al.  The role of pericytes in blood-vessel formation and maintenance. , 2005, Neuro-oncology.

[36]  C. Betsholtz,et al.  Pericytes and the blood-brain barrier: recent advances and implications for the delivery of CNS therapy. , 2011, Therapeutic delivery.

[37]  M. Santoro,et al.  An α-Smooth Muscle Actin (acta2/αsma) Zebrafish Transgenic Line Marking Vascular Mural Cells and Visceral Smooth Muscle Cells , 2014, PloS one.

[38]  S. Strickland,et al.  Ablation of astrocytic laminin impairs vascular smooth muscle cell function and leads to hemorrhagic stroke , 2013, The Journal of cell biology.

[39]  Chih‐Chung Lin,et al.  TNF-α induces MMP-9 expression via activation of Src/EGFR, PDGFR/PI3K/Akt cascade and promotion of NF-κB/p300 binding in human tracheal smooth muscle cells , 2007 .

[40]  J. Gutkind,et al.  Assembly and patterning of the vascular network of the vertebrate hindbrain , 2011, Development.

[41]  F. Pieruzzi,et al.  ANG II increases TIMP-1 expression in rat aortic smooth muscle cells in vivo. , 2003, American journal of physiology. Heart and circulatory physiology.

[42]  B. Lilly,et al.  Transforming Growth Factor-β (TGF-β1) Down-regulates Notch3 in Fibroblasts to Promote Smooth Muscle Gene Expression* , 2008, Journal of Biological Chemistry.

[43]  J. Campos-Ortega,et al.  Use of the Gal4-UAS technique for targeted gene expression in the zebrafish , 1999, Mechanisms of Development.

[44]  K. L. de Mesy Bentley,et al.  Ultrastructure of zebrafish dorsal aortic cells. , 2006, Zebrafish.

[45]  B. Barres,et al.  Pericytes are required for blood–brain barrier integrity during embryogenesis , 2010, Nature.

[46]  G. Nixon,et al.  Sphingosine-1-phosphate-induced release of TIMP-2 from vascular smooth muscle cells inhibits angiogenesis , 2012, Journal of Cell Science.

[47]  B. Lilly,et al.  Notch2 and Notch3 Function Together to Regulate Vascular Smooth Muscle Development , 2012, PloS one.

[48]  C. Moens,et al.  Notch3 establishes brain vascular integrity by regulating pericyte number , 2014, Development.

[49]  Amber N. Stratman,et al.  Endothelial Cell-Pericyte Interactions Stimulate Basement Membrane Matrix Assembly: Influence on Vascular Tube Remodeling, Maturation, and Stabilization , 2011, Microscopy and Microanalysis.

[50]  B. Lilly,et al.  MicroRNA miR145 Regulates TGFBR2 Expression and Matrix Synthesis in Vascular Smooth Muscle Cells , 2015, Circulation research.

[51]  A. Eichmann,et al.  Vascular Mural Cells Promote Noradrenergic Differentiation of Embryonic Sympathetic Neurons. , 2015, Cell reports.

[52]  N. Fusenig,et al.  Inhibition of basement membrane formation by a nidogen-binding laminin γ1-chain fragment in human skin-organotypic cocultures , 2004, Journal of Cell Science.

[53]  D. Stainier,et al.  Characterization of vascular mural cells during zebrafish development , 2009, Mechanisms of Development.

[54]  B. Weinstein,et al.  Angiogenic network formation in the developing vertebrate trunk , 2003, Development.

[55]  S. Strickland,et al.  Astrocytic laminin regulates pericyte differentiation and maintains blood brain barrier integrity , 2014, Nature Communications.

[56]  R. Adams,et al.  Integrin &bgr;1 Subunit Controls Mural Cell Adhesion, Spreading, and Blood Vessel Wall Stability , 2008, Circulation research.

[57]  K. Hirschi,et al.  Gap Junction Communication Mediates Transforming Growth Factor-&bgr; Activation and Endothelial-Induced Mural Cell Differentiation , 2003, Circulation research.

[58]  C. Betsholtz,et al.  Reverse genetic screening reveals poor correlation between morpholino-induced and mutant phenotypes in zebrafish. , 2015, Developmental cell.

[59]  C. Kimmel,et al.  Stages of embryonic development of the zebrafish , 1995, Developmental dynamics : an official publication of the American Association of Anatomists.

[60]  K. Pottin,et al.  Sclerotomal origin of vascular smooth muscle cells and pericytes in the embryo. , 2008, Developmental biology.

[61]  Cheol‐Hee Kim,et al.  Four twist genes in zebrafish, four expression patterns , 2007, Developmental dynamics : an official publication of the American Association of Anatomists.

[62]  J. Partanen,et al.  Developmental origin of smooth muscle cells in the descending aorta in mice , 2008, Development.

[63]  Amber N. Stratman,et al.  Pericyte recruitment during vasculogenic tube assembly stimulates endothelial basement membrane matrix formation. , 2009, Blood.

[64]  Neville E Sanjana,et al.  A transcription activator-like effector toolbox for genome engineering , 2012, Nature Protocols.

[65]  G. Davis,et al.  Coregulation of vascular tube stabilization by endothelial cell TIMP-2 and pericyte TIMP-3 , 2006, The Journal of cell biology.

[66]  Marcus Krüger,et al.  Genetic compensation induced by deleterious mutations but not gene knockdowns , 2015, Nature.

[67]  Chih‐Chung Lin,et al.  TNF-alpha induces MMP-9 expression via activation of Src/EGFR, PDGFR/PI3K/Akt cascade and promotion of NF-kappaB/p300 binding in human tracheal smooth muscle cells. , 2007, American journal of physiology. Lung cellular and molecular physiology.

[68]  R. Hynes.  Cell–matrix adhesion in vascular development , 2007, Journal of thrombosis and haemostasis : JTH.

[69]  Nico Stuurman,et al.  Computer Control of Microscopes Using µManager , 2010, Current protocols in molecular biology.

[70]  Rakesh K Jain,et al.  Molecular regulation of vessel maturation , 2003, Nature Medicine.

[71]  C. Betsholtz,et al.  Role of PDGF-B and PDGFR-beta in recruitment of vascular smooth muscle cells and pericytes during embryonic blood vessel formation in the mouse. , 1999, Development.