Eya1 controls cell polarity, spindle orientation, cell fate and Notch signaling in distal embryonic lung epithelium

Cell polarity, mitotic spindle orientation and asymmetric division play a crucial role in the self-renewal/differentiation of epithelial cells, yet little is known about these processes and the molecular programs that control them in embryonic lung distal epithelium. Herein, we provide the first evidence that embryonic lung distal epithelium is polarized with characteristic perpendicular cell divisions. Consistent with these findings, spindle orientation-regulatory proteins Insc, LGN (Gpsm2) and NuMA, and the cell fate determinant Numb are asymmetrically localized in embryonic lung distal epithelium. Interfering with the function of these proteins in vitro randomizes spindle orientation and changes cell fate. We further show that Eya1 protein regulates cell polarity, spindle orientation and the localization of Numb, which inhibits Notch signaling. Hence, Eya1 promotes both perpendicular division as well as Numb asymmetric segregation to one daughter in mitotic distal lung epithelium, probably by controlling aPKCζ phosphorylation. Thus, epithelial cell polarity and mitotic spindle orientation are defective after interfering with Eya1 function in vivo or in vitro. In addition, in Eya1−/− lungs, perpendicular division is not maintained and Numb is segregated to both daughter cells in mitotic epithelial cells, leading to inactivation of Notch signaling. As Notch signaling promotes progenitor cell identity at the expense of differentiated cell phenotypes, we test whether genetic activation of Notch could rescue the Eya1−/− lung phenotype, which is characterized by loss of epithelial progenitors, increased epithelial differentiation but reduced branching. Indeed, genetic activation of Notch partially rescues Eya1−/− lung epithelial defects. These findings uncover novel functions for Eya1 as a crucial regulator of the complex behavior of distal embryonic lung epithelium.

[1]  D. Warburton,et al.  Eyes absent 1 (Eya1) is a critical coordinator of epithelial, mesenchymal and vascular morphogenesis in the mammalian lung. , 2011, Developmental biology.

[2]  Jing Liu,et al.  LGN regulates mitotic spindle orientation during epithelial morphogenesis , 2010, The Journal of cell biology.

[3]  B. Hogan,et al.  The Id2+ distal tip lung epithelium contains individual multipotent embryonic progenitor cells , 2009, Development.

[4]  D. Guidolin,et al.  miR-17 family of microRNAs controls FGF10-mediated embryonic lung epithelial branching morphogenesis through MAPK14 and STAT3 regulation of E-Cadherin distribution. , 2009, Developmental biology.

[5]  F. Matsuzaki,et al.  Protein phosphatase 2A negatively regulates aPKC signaling by modulating phosphorylation of Par-6 in Drosophila neuroblast asymmetric divisions , 2009, Journal of Cell Science.

[6]  S. Li,et al.  Numb regulates cell–cell adhesion and polarity in response to tyrosine kinase signalling , 2009, The EMBO journal.

[7]  P. Tsao,et al.  Notch signaling controls the balance of ciliated and secretory cell fates in developing airways , 2009, Development.

[8]  B. Ang,et al.  Protein phosphatase 2A regulates self-renewal of Drosophila neural stem cells , 2009, Development.

[9]  D. Melton,et al.  Notch signaling promotes airway mucous metaplasia and inhibits alveolar development , 2009, Development.

[10]  Chris Q. Doe,et al.  Spindle orientation during asymmetric cell division , 2009, Nature Cell Biology.

[11]  Christopher K. Glass,et al.  Tyrosine Dephosphorylation of H2AX Modulates Apoptosis and Survival Decisions , 2009, Nature.

[12]  I. Rebay,et al.  Interactions with the Abelson tyrosine kinase reveal compartmentalization of eyes absent function between nucleus and cytoplasm. , 2009, Developmental cell.

[13]  B. Ang,et al.  Protein phosphatase 2 A regulates self-renewal of Drosophila neural stem cells , 2009 .

[14]  Bernd Fritzsch,et al.  Eya1 gene dosage critically affects the development of sensory epithelia in the mammalian inner ear. , 2008, Human molecular genetics.

[15]  P. Tsao,et al.  γ-Secretase Activation of Notch Signaling Regulates the Balance of Proximal and Distal Fates in Progenitor Cells of the Developing Lung* , 2008, Journal of Biological Chemistry.

[16]  E. Rawlins.  Lung epithelial progenitor cells: lessons from development. , 2008, Proceedings of the American Thoracic Society.

[17]  M. Klymkowsky,et al.  Eya1 and Six1 promote neurogenesis in the cranial placodes in a SoxB1-dependent fashion. , 2008, Developmental biology.

[18]  Ophir D. Klein,et al.  The branching programme of mouse lung development , 2008, Nature.

[19]  D. Warburton.  Developmental biology: Order in the lung , 2008, Nature.

[20]  Yun Lu,et al.  Epithelial progenitor cells of the embryonic lung and the role of microRNAs in their proliferation. , 2008, Proceedings of the American Thoracic Society.

[21]  N. Gaiano,et al.  Differential Notch signalling distinguishes neural stem cells from intermediate progenitors. , 2007, Nature.

[22]  K. Kaibuchi,et al.  Numb controls integrin endocytosis for directional cell migration with aPKC and PAR-3. , 2007, Developmental cell.

[23]  I. Rebay,et al.  The eyes absent family of phosphotyrosine phosphatases: properties and roles in developmental regulation of transcription. , 2007, Annual review of biochemistry.

[24]  N. Mackman,et al.  TLR2 Is Constitutively Expressed within the Kidney and Participates in Ischemic Renal Injury through Both MyD88-Dependent and -Independent Pathways1 , 2007, The Journal of Immunology.

[25]  J. Casanova.  PARtitioning Numb , 2007, EMBO reports.

[26]  P. Thibault,et al.  aPKC‐mediated phosphorylation regulates asymmetric membrane localization of the cell fate determinant Numb , 2007, The EMBO journal.

[27]  S. Shapiro,et al.  Stage-specific Effects of cAMP Signaling during Distal Lung Epithelial Development* , 2006, Journal of Biological Chemistry.

[28]  C. Cepko,et al.  Notch activity permits retinal cells to progress through multiple progenitor states and acquire a stem cell property , 2006, Proceedings of the National Academy of Sciences.

[29]  U. Lendahl,et al.  High levels of Notch signaling down-regulate Numb and Numblike , 2006, The Journal of cell biology.

[30]  H. Rockoff.  On the Origins of , 2006 .

[31]  D. Siderovski,et al.  Dynamic regulation of mammalian numb by G protein-coupled receptors and protein kinase C activation: Structural determinants of numb association with the cortical membrane. , 2006, Molecular biology of the cell.

[32]  D. Warburton,et al.  Differential role of FGF9 on epithelium and mesenchyme in mouse embryonic lung. , 2006, Developmental biology.

[33]  A. Suzuki,et al.  The PAR-aPKC system: lessons in polarity , 2006, Journal of Cell Science.

[34]  D. Warburton,et al.  VEGF-A signaling through Flk-1 is a critical facilitator of early embryonic lung epithelial to endothelial crosstalk and branching morphogenesis. , 2006, Developmental biology.

[35]  Elaine Fuchs,et al.  Asymmetric cell divisions promote stratification and differentiation of mammalian skin , 2005, Nature.

[36]  J. Knoblich,et al.  Numb and α‐Adaptin regulate Sanpodo endocytosis to specify cell fate in Drosophila external sensory organs , 2005, EMBO reports.

[37]  D. Warburton,et al.  A novel function for the protein tyrosine phosphatase Shp2 during lung branching morphogenesis. , 2005, Developmental biology.

[38]  D. Warburton,et al.  In vivo inosine protects alveolar epithelial type 2 cells against hyperoxia-induced DNA damage through MAP kinase signaling. , 2005, American journal of physiology. Lung cellular and molecular physiology.

[39]  Bernd Fritzsch,et al.  Eya1 and Six1 are essential for early steps of sensory neurogenesis in mammalian cranial placodes , 2004, Development.

[40]  Donald G Welsh,et al.  Cell-cell interactions in regulating lung function. , 2004, American journal of physiology. Lung cellular and molecular physiology.

[41]  J. Knoblich,et al.  Dare to Be Different: Asymmetric Cell Division in Drosophila, C. elegans and Vertebrates , 2004, Current Biology.

[42]  S. Krauss,et al.  Continuing role for mouse Numb and Numbl in maintaining progenitor cells during cortical neurogenesis , 2004, Nature Neuroscience.

[43]  R. Klein,et al.  Notch activation induces apoptosis in neural progenitor cells through a p53-dependent pathway. , 2004, Developmental biology.

[44]  T. Golub,et al.  The Six1 homeoprotein stimulates tumorigenesis by reactivation of cyclin A1. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[45]  I. Macara.  Par Proteins: Partners in Polarization , 2004, Current Biology.

[46]  A. E. Bishop.  Pulmonary epithelial stem cells , 2004, Cell proliferation.

[47]  A. Kriegstein,et al.  Cortical neurons arise in symmetric and asymmetric division zones and migrate through specific phases , 2004, Nature Neuroscience.

[48]  T. Golub,et al.  The Six 1 homeoprotein stimulates tumorigenesis by reactivation of cyclin A 1 , 2004 .

[49]  J. Beckmann,et al.  Six and Eya expression during human somitogenesis and MyoD gene family activation , 2004, Journal of Muscle Research & Cell Motility.

[50]  Claudia S. Barros,et al.  Drosophila nonmuscle myosin II promotes the asymmetric segregation of cell fate determinants by cortical exclusion rather than active transport. , 2003, Developmental cell.

[51]  C. Glass,et al.  Eya protein phosphatase activity regulates Six1–Dach–Eya transcriptional effects in mammalian organogenesis , 2003, Nature.

[52]  J. Epstein,et al.  Signal transduction: An eye on organ development , 2003, Nature.

[53]  W. Nelson,et al.  Epithelial cell polarity from the outside looking in. , 2003, News in physiological sciences : an international journal of physiology produced jointly by the International Union of Physiological Sciences and the American Physiological Society.

[54]  U. Tepass.  Claudin complexities at the apical junctional complex , 2003, Nature Cell Biology.

[55]  M. Raff,et al.  The orientation of cell division influences cell-fate choice in the developing mammalian retina , 2003, Development.

[56]  W. Nelson,et al.  Adaptation of core mechanisms to generate cell polarity , 2003, Nature.

[57]  Pasko Rakic,et al.  Mitotic spindle rotation and mode of cell division in the developing telencephalon , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[58]  Martin Raff,et al.  Asymmetric segregation of Numb: a mechanism for neural specification from Drosophila to mammals , 2002, Nature Neuroscience.

[59]  Y. Jan,et al.  Progenitor cell maintenance requires numb and numblike during mouse neurogenesis , 2002, Nature.

[60]  Sally Temple,et al.  Asymmetric Numb distribution is critical for asymmetric cell division of mouse cerebral cortical stem cells and neuroblasts. , 2002, Development.

[61]  D. Warburton,et al.  mSprouty2 inhibits FGF10-activated MAP kinase by differentially binding to upstream target proteins. , 2002, American journal of physiology. Lung cellular and molecular physiology.

[62]  C. White,et al.  Protein phosphatase 2A associates with and regulates atypical PKC and the epithelial tight junction complex , 2002, The Journal of cell biology.

[63]  Susan E. Wert,et al.  Early restriction of peripheral and proximal cell lineages during formation of the lung , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[64]  R. Maas,et al.  Eya1 is required for the morphogenesis of mammalian thymus, parathyroid and thyroid. , 2002, Development.

[65]  C. Guidos,et al.  Transgenic Expression of Numb Inhibits Notch Signaling in Immature Thymocytes But Does Not Alter T Cell Fate Specification1 , 2002, The Journal of Immunology.

[66]  M. Prentki,et al.  Protein Kinase Cζ Activation Mediates Glucagon-Like Peptide-1–Induced Pancreatic β-Cell Proliferation , 2001 .

[67]  S. Ohno.  Intercellular junctions and cellular polarity: the PAR-aPKC complex, a conserved core cassette playing fundamental roles in cell polarity. , 2001, Current opinion in cell biology.

[68]  M. Raff,et al.  Asymmetric Segregation of Numb in Retinal Development and the Influence of the Pigmented Epithelium , 2001, The Journal of Neuroscience.

[69]  B. Hogan,et al.  Notch/Delta expression in the developing mouse lung , 2000, Mechanisms of Development.

[70]  W. Cardoso.  Lung morphogenesis revisited: Old facts, current ideas , 2000, Developmental dynamics : an official publication of the American Association of Anatomists.

[71]  L. Shoshani,et al.  Molecular physiology and pathophysiology of tight junctions. I. Biogenesis of tight junctions and epithelial polarity. , 2000, American journal of physiology. Gastrointestinal and liver physiology.

[72]  K. Mostov,et al.  Membrane traffic in polarized epithelial cells. , 2000, Current opinion in cell biology.

[73]  David Warburton,et al.  The molecular basis of lung morphogenesis , 2000, Mechanisms of Development.

[74]  P. Cohen,et al.  Specificity and mechanism of action of some commonly used protein kinase inhibitors. , 2000, The Biochemical journal.

[75]  Joe C. Adams,et al.  Eya1-deficient mice lack ears and kidneys and show abnormal apoptosis of organ primordia , 1999, Nature Genetics.

[76]  J. Brody,et al.  FGF-2 induces surfactant protein gene expression in foetal rat lung epithelial cells through a MAPK-independent pathway. , 1999, Cellular signalling.

[77]  A. Newton,et al.  Regulation of conventional protein kinase C isozymes by phosphoinositide-dependent kinase 1 (PDK-1) , 1998, Current Biology.

[78]  H. Ford,et al.  Abrogation of the G2 cell cycle checkpoint associated with overexpression of HSIX1: a possible mechanism of breast carcinogenesis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[79]  H. Yin,et al.  The Yeast Spindle Pole Body Component Spc72p Interacts with Stu2p and Is Required for Proper Microtubule Assembly , 1998, The Journal of cell biology.

[80]  J. Epstein,et al.  Mouse Eya genes are expressed during limb tendon development and encode a transcriptional activation function. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[81]  R. Maas,et al.  Mouse Eya homologues of the Drosophila eyes absent gene require Pax6 for expression in lens and nasal placode. , 1997, Development.

[82]  Y. Jan,et al.  Control of Daughter Cell Fates during Asymmetric Division: Interaction of Numb and Notch , 1996, Neuron.

[83]  D. Drubin,et al.  Origins of Cell Polarity , 1996, Cell.

[84]  Y. Jan,et al.  Asymmetric segregation of Numb and Prospero during cell division , 1995, Nature.

[85]  A. Newton,et al.  In vivo regulation of protein kinase C by trans-phosphorylation followed by autophosphorylation. , 1994, The Journal of biological chemistry.

[86]  Y. Jan,et al.  Asymmetric distribution of numb protein during division of the sensory organ precursor cell confers distinct fates to daughter cells , 1994, Cell.

[87]  M. Williams,et al.  An improved method for isolating type II cells in high yield and purity. , 2015, The American review of respiratory disease.

[88]  W. Crocker,et al.  Polarity , 1910, Botanical Gazette.

[89]  RosaGFP SPCCre.  Notch signaling promotes airway mucous metaplasia and inhibits alveolar development , 2022 .