A Quantitative Cell Migration Assay for Murine Enteric Neural Progenitors

Neural crest cells (NCC) are a transient and multipotent cell population that originates from the dorsal neural tube and migrates extensively throughout ...

UQAM

Although the development of sea urchin embryos has been studied extensively and clearly involves both cell adhesion and cell migration, rather little is known about the adhesion receptors and extracellular matrix molecules involved. The completion of the genome of Strongylocentrotus purpuratus allows a comprehensive survey of the complement of cell-cell and cell-matrix adhesion molecules in this organism. Furthermore, the phylogenetic position of echinoderms offers the opportunity to compare the complement of adhesion proteins between protostome and deuterostome invertebrates and between invertebrate and vertebrate deuterostomes. Many aspects of development and cell interactions differ among these different taxa and it is likely that analysis of the spectrum of adhesion receptors and extracellular matrix proteins can open up new insights into which molecules have evolved to suit particular developmental processes. In this paper, we report the results of an initial analysis along these lines. The echinoderm adhesome (complement of adhesion-related genes/proteins) is similar overall to that of other invertebrates although there are significant deuterostome-specific innovations and some interesting features previously thought to be chordate or vertebrate specific.

The echinoderm adhesome.

Wir berichten die Abfolge und die Analyse des Genoms 814-Megabase Seeigel Strongylocentrotus Purpuratus, ein Modell f&uuml;r Entwicklungsbiologie und Systembiologie. Die Sequenzierung-Strategie kombiniert ganz-Genoms Schrotflinte und bacterial artificial Chromosome (BAC) Sequenzen. Diese Verwendung von BAC Klone, unterst&uuml;tzt durch ein pooling-Strategie, &uuml;berwand die Schwierigkeiten im Zusammenhang mit hohen Heterozygosit&auml;t des Genoms. Das Genom kodiert rund 23.300 Gene, darunter viele, die vorher als Wirbeltiere Innovationen werden oder nur au&szlig;erhalb der Deuterostomiern bekannt. Diese Stachelh&auml;uter-Genom enth&auml;lt einen evolution&auml;ren Fremdgruppe f&uuml;r die Chordatiere und Renditen Einblicke in die Entwicklung der Deuterostomiern.

Das Genom der Seeigel Strongylocentrotus Purpuratus.

Riportiamo la sequenza e l&#39;analisi del genoma 814-megabase di riccio di mare Strongylocentrotus purpuratus, un modello per lo sviluppo e la biologia dei sistemi. La strategia di sequenziamento combinato whole genome shotgun e sequenze di cromosomi artificiali batterici (BAC). Questo uso di cloni BAC, aiutato da una strategia di pooling, superato Difficolt&agrave; associate con elevata eterozigosi del genoma. Il genoma codifica circa 23.300 geni, tra cui molti pensava di essere vertebrate innovazioni o noto solo di fuori i deuterostomi. Questo genoma echinodermi fornisce un evolutivo outgroup per i cordati e rendimenti intuizioni l&#39;evoluzione dei deuterostomi.

Il genoma del riccio di mare Strongylocentrotus purpuratus.

&#x6211;&#x4EEC;&#x62A5;&#x544A;&#x7684;&#x987A;&#x5E8F;&#x548C;&#x6D77;&#x80C6;&#x6D77;&#x80C6; purpuratus&#xFF0C;&#x4E00;&#x79CD;&#x6A21;&#x578B;&#x7B2C; 814 &#x7ECF;&#x8FC7;&#x57FA;&#x56E0;&#x7EC4;&#x5206;&#x6790;&#x53D1;&#x5C55;&#x548C;&#x7CFB;&#x7EDF;&#x751F;&#x7269;&#x5B66;&#x3002;&#x6D4B;&#x5E8F;&#x6218;&#x7565;&#x8054;&#x5408;&#x5168;&#x57FA;&#x56E0;&#x7EC4;&#x9E1F;&#x67AA;&#x548C;&#x7EC6;&#x83CC;&#x4EBA;&#x5DE5;&#x67D3;&#x8272;&#x4F53; (BAC) &#x5E8F;&#x5217;&#x3002;&#x8FD9;&#x79CD;&#x5229;&#x7528;&#x6240;&#x6C47;&#x96C6;&#x7684;&#x7B56;&#x7565;&#xFF0C;&#x8F85;&#x52A9;&#x7684; BAC &#x514B;&#x9686;&#x514B;&#x670D;&#x4E86;&#x56F0;&#x96BE;&#x4E0E;&#x9AD8;&#x6742;&#x5408;&#x6027;&#x57FA;&#x56E0;&#x7EC4;&#x76F8;&#x5173;&#x8054;&#x3002;&#x57FA;&#x56E0;&#x7EC4;&#x7F16;&#x7801;&#x7EA6; 23,300 &#x7684;&#x57FA;&#x56E0;&#xFF0C;&#x5305;&#x62EC;&#x8BB8;&#x591A;&#x4EE5;&#x524D;&#x8BA4;&#x4E3A;&#x662F;&#x810A;&#x690E;&#x52A8;&#x7269;&#x521B;&#x65B0;&#x6216; deuterostomes &#x4EE5;&#x5916;&#x53EA;&#x77E5;&#x9053;&#x3002;&#x8FD9;&#x4E2A;&#x68D8;&#x76AE;&#x52A8;&#x7269;&#x57FA;&#x56E0;&#x7EC4;&#x63D0;&#x4F9B;&#x4E86;&#x8FDB;&#x5316; outgroup &#x5230; deuterostomes &#x7684;&#x6F14;&#x53D8;&#x810A;&#x7D22;&#x52A8;&#x7269;&#x548C;&#x6536;&#x76CA;&#x7387;&#x7684;&#x6DF1;&#x523B;&#x89C1;&#x89E3;&#x3002;

&#x6D77;&#x80C6;&#x6D77;&#x80C6; purpuratus &#x7684;&#x57FA;&#x56E0;&#x7EC4;&#x3002;

We report the sequence and analysis of the 814-megabase genome of the sea urchin Strongylocentrotus purpuratus, a model for developmental and systems biology. The sequencing strategy combined whole-genome shotgun and bacterial artificial chromosome (BAC) sequences. This use of BAC clones, aided by a pooling strategy, overcame difficulties associated with high heterozygosity of the genome. The genome encodes about 23,300 genes, including many previously thought to be vertebrate innovations or known only outside the deuterostomes. This echinoderm genome provides an evolutionary outgroup for the chordates and yields insights into the evolution of deuterostomes.

The genome of the sea urchin Strongylocentrotus purpuratus.

Nous rapportons la s&eacute;quence et l&#39;analyse du g&eacute;nome de l&#39;oursin Strongylocentrotus purpuratus, un mod&egrave;le pour 814-megabase d&eacute;veloppementale et biologie des syst&egrave;mes. La strat&eacute;gie de s&eacute;quen&ccedil;age combin&eacute; whole-genome shotgun et des s&eacute;quences des chromosomes artificiels bact&eacute;riens (BAC). Cette utilisation des clones BAC, aid&eacute; par une strat&eacute;gie de mise en commun, a surmont&eacute; les difficult&eacute;s associ&eacute;es &agrave; l&#39;h&eacute;t&eacute;rozygotie &eacute;lev&eacute; du g&eacute;nome. Le g&eacute;nome encode environ 23 300 g&egrave;nes, dont un grand nombre qu&#39;on croyait &ecirc;tre des innovations vert&eacute;br&eacute;es ou connu seulement en dehors des deut&eacute;rostomiens. Ce g&eacute;nome &eacute;chinoderme fournit un &eacute;volutionniste exogroupe pour les id&eacute;es chord&eacute;s et les rendements dans l&#39;&eacute;volution des deut&eacute;rostomiens.

Le g&eacute;nome de l&#39;oursin Strongylocentrotus purpuratus.

&#x6211;&#x3005; &#x306F;&#x3001;&#x30B7;&#x30FC;&#x30B1;&#x30F3;&#x30B9;&#x3068;&#x30A6;&#x30CB; &#x30C8;&#x30E9;&#x30B6;&#x30E1; purpuratus &#x306E;&#x305F;&#x3081;&#x306E;&#x30E2;&#x30C7;&#x30EB;&#x306E; 814 &#x7D50;&#x679C;&#x30B2;&#x30CE;&#x30E0;&#x89E3;&#x6790;&#x5831;&#x544A;&#x767A;&#x9054;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x751F;&#x7269;&#x5B66;&#x3002;&#x30B7;&#x30FC;&#x30B1;&#x30F3;&#x30B9;&#x306E;&#x6226;&#x7565;&#x306F;&#x3001;&#x5168;&#x30B2;&#x30CE;&#x30E0;&#x306E;&#x6563;&#x5F3E;&#x9283;&#x3068;&#x7D30;&#x83CC;&#x4EBA;&#x5DE5;&#x67D3;&#x8272;&#x4F53; &#xFF08;BAC &#x306E;&#xFF09; &#x30B7;&#x30FC;&#x30B1;&#x30F3;&#x30B9;&#x7D50;&#x5408;&#x3002;&#x3053;&#x306E;&#x4F7F;&#x7528;&#x30D7;&#x30FC;&#x30EA;&#x30F3;&#x30B0;&#x306E;&#x6226;&#x7565;&#x306B;&#x3088;&#x3063;&#x3066;&#x3001;&#x652F;&#x63F4;&#x3001;BAC &#x30AF;&#x30ED;&#x30FC;&#x30F3;&#x306E;&#x30B2;&#x30CE;&#x30E0;&#x306E;&#x9AD8;&#x3044;&#x30D8;&#x30C6;&#x30ED;&#x63A5;&#x5408;&#x6027;&#x3068;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x308B;&#x96E3;&#x3057;&#x3055;&#x3092;&#x514B;&#x670D;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x30B2;&#x30CE;&#x30E0;&#x4EE5;&#x524D;&#x810A;&#x690E;&#x52D5;&#x7269;&#x306E;&#x6280;&#x8853;&#x9769;&#x65B0;&#x3067;&#x3042;&#x308B;&#x8003;&#x3048;&#x307E;&#x305F;&#x306F;&#x306E;&#x307F;&#x3001;&#x5F8C;&#x53E3;&#x52D5;&#x7269;&#x5916;&#x77E5;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x591A;&#x304F;&#x3092;&#x542B;&#x3080;&#x3001;&#x7D04; 23,300 &#x907A;&#x4F1D;&#x5B50;&#x3092;&#x30A8;&#x30F3;&#x30B3;&#x30FC;&#x30C9;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x68D8;&#x76AE;&#x52D5;&#x7269;&#x306E;&#x30B2;&#x30CE;&#x30E0;&#x9032;&#x5316;&#x63D0;&#x4F9B;&#x5F8C;&#x53E3;&#x52D5;&#x7269;&#x306E;&#x9032;&#x5316;&#x306E;&#x810A;&#x7D22;&#x52D5;&#x7269;&#x3068;&#x5229;&#x56DE;&#x308A;&#x6D1E;&#x5BDF;&#x3092;&#x91CD;&#x8981;&#x3002;

&#x30A6;&#x30CB; &#x30C8;&#x30E9;&#x30B6;&#x30E1; purpuratus &#x306E;&#x30B2;&#x30CE;&#x30E0;&#x3002;

&#xC6B0;&#xB9AC;&#xB294; &#xC2DC;&#xD000;&#xC2A4; &#xBC0F; &#xC131; Strongylocentrotus purpuratus&#xC5D0; &#xB300; &#xD55C; &#xBAA8;&#xB378;&#xC758; 814 megabase &#xAC8C;&#xB188; &#xBD84;&#xC11D; &#xBCF4;&#xACE0;&#xC11C; &#xAC1C;&#xBC1C; &#xBC0F; &#xC2DC;&#xC2A4;&#xD15C; &#xC0DD;&#xBB3C;&#xD559;. &#xC2DC;&#xD000;&#xC2F1; &#xC804;&#xB7B5; &#xC804;&#xCCB4; &#xAC8C;&#xB188; &#xC0F7;&#xAC74; &#xD558; &#xACE0; &#xC138;&#xADE0; &#xC778;&#xACF5; &#xC5FC;&#xC0C9;&#xCCB4; (BAC) &#xC2DC;&#xD000;&#xC2A4; &#xACB0;&#xD569;. BAC &#xD074;&#xB860;, &#xD480;&#xB9C1; &#xC804;&#xB7B5;&#xC5D0; &#xC758;&#xD574; &#xC6D0;&#xC870;&#xC758;&#xC774; &#xC0AC;&#xC6A9;&#xC774; &#xAC8C;&#xB188;&#xC758; &#xB192;&#xC740; heterozygosity&#xC640; &#xAD00;&#xB828; &#xB41C; &#xC5B4;&#xB824;&#xC6C0;&#xC744; &#xADF9;&#xBCF5; &#xD588;&#xB2E4;. &#xAC8C;&#xB188;&#xC5D0;&#xB294; &#xC57D; 23,300 &#xC720;&#xC804;&#xC790;&#xB97C; &#xD3EC;&#xD568; &#xD55C; &#xB9CE;&#xC740; &#xC774;&#xC804;&#xC5D0; &#xCC99;&#xCD94; &#xD601;&#xC2E0; &#xC0DD;&#xAC01; &#xB098;&#xB294; deuterostomes &#xBC16;&#xC5D0; &#xC11C;&#xB9CC; &#xC54C;&#xB824;&#xC9C4; &#xC778;&#xCF54;&#xB529;&#xD569;&#xB2C8;&#xB2E4;. &#xADF9;&#xD53C;&#xB3D9;&#xBB3C; &#xAC8C;&#xB188;&#xC774; &#xC81C;&#xACF5; &#xD558;&#xB294; &#xC9C4;&#xD654; outgroup chordates &#xBC0F; &#xC218;&#xC728; &#xD1B5;&#xCC30;&#xB825; deuterostomes&#xC758; &#xC9C4;&#xD654;&#xC5D0; &#xB300; &#xD55C;.

&#xBC14;&#xB2E4; &#xC131; &#xAC8C; Strongylocentrotus purpuratus&#xC758; &#xAC8C;&#xB188;&#xC785;&#xB2C8;&#xB2E4;.

Divulgamos la secuencia y el an&aacute;lisis del genoma 814-megabase del erizo de mar Strongylocentrotus purpuratus, un modelo de desarrollo y Biolog&iacute;a de sistemas. La estrategia de secuenciaci&oacute;n hab&iacute;a combinado escopeta todo el genoma y secuencias del cromosoma artificial bacteriano (BAC). Este uso de clones BAC, ayudado por una estrategia de agrupaci&oacute;n super&oacute; dificultades asociadas con el alto heterocigoto del genoma. El genoma codifica unos 23.300 genes, incluyendo muchos pensaba ser vertebradas innovaciones o conocida s&oacute;lo fuera los deuterostomes. Este genoma de equinodermos proporciona un evolutivo de grupos para las penetraciones de cordados y rendimientos en la evoluci&oacute;n de deuterostomes.

El genoma del erizo de mar Strongylocentrotus purpuratus.

Glycosaminoglycans (GAGs) are a heavily sulfated component of the extracellular matrix (ECM) implicated in a variety of cell signaling events involved in patterning of embryos. Embryos of the sea urchin Strongylocentrotus purpuratus were exposed to several inhibitors that disrupt GAG function during development. Treatment with chlorate, a general inhibitor of sulfation that leads to undersulfated GAGs, reduced sulfation of the urchin blastocoelar ECM. It also prevented correct specification of the oral-aboral axis and mouth formation, resulting in a radialized phenotype characterized by the lack of an oral field, incomplete gastrulation and formation of multiple skeletal spicule rudiments. Oral markers were initially expressed in most of the prospective ectoderm of chlorate-treated early blastulae, but then declined as aboral markers became expressed throughout most of the ectoderm. Nodal expression in the presumptive oral field is necessary and sufficient to specify the oral-aboral axis in urchins. Several lines of evidence suggest a deregulation of Nodal signaling is involved in the radialization caused by chlorate: (1) Radial embryos resemble those in which Nodal expression was knocked down. (2) Chlorate disrupted localized nodal expression in oral ectoderm, even when applied after the oral-aboral axis is specified and expression of other oral markers is resistant to treatment. (3) Inhibition with SB-431542 of ALK-4/5/7 receptors that mediate Nodal signaling causes defects in ectodermal patterning similar to those caused by chlorate. (4) Intriguingly, treatment of embryos with a sub-threshold dose of SB-431542 rescued the radialization caused by low concentrations of chlorate. Our results indicate important roles for sulfated GAGs in Nodal signaling and oral-aboral axial patterning, and in the cellular processes necessary for archenteron extension and mouth formation during gastrulation. We propose that interaction of the Nodal ligand with sulfated GAGs limits its diffusion, and is required to specify an oral field in the urchin embryo and organize the oral-aboral axis.

Karl-Frederik Bergeron

Molecular Genetics of Development Laboratory,
Biological Sciences,
Molecular Genetics of Development Laboratory, Biological Sciences

UQAM

A Quantitative Cell Migration Assay for Murine Enteric Neural Progenitors

Karl-Frederik Bergeron

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Molecular Genetics of Development Laboratory,Biological Sciences,Molecular Genetics of Development Laboratory, Biological Sciences

UQAM

A Quantitative Cell Migration Assay for Murine Enteric Neural Progenitors

Molecular Genetics of Development Laboratory,
Biological Sciences,
Molecular Genetics of Development Laboratory, Biological Sciences