visualization-craniofacial-development-sox10-kaede-transgenic

Visualization of Craniofacial Development in the sox10: kaede Transgenic Zebrafish Line Using Time-lapse Confocal Microscopy

Vertebrate palatogenesis is a highly choreographed and complex developmental process, which involves migration of cranial neural crest (CNC) cells, ...

Massachusetts General Hospital

Cranial neural crest cells follow stereotypic patterns of migration to form craniofacial structures. The zebrafish is a powerful vertebrate genetic model where transgenics with reporter proteins under the transcriptional regulation of lineage-specific promoters can be generated. Numerous studies demonstrate that the zebrafish ethmoid plate is embryologically analogous to the mammalian palate. A fate map correlating embryonic cranial neural crest to defined jaw structures would provide a useful context for the morphogenetic analysis of craniofacial development. To that end, the sox10:kaede transgenic was generated, where sox10 provides lineage restriction to the neural crest. Specific regions of neural crest were labeled at the 10-somite stage by photoconversion of the kaede reporter protein. Lineage analysis was carried out during pharyngeal development in wild-type animals, after miR140 injection, and after estradiol treatment. At the 10-somite stage, cranial neural crest cells anterior of the eye contributed to the median ethmoid plate, whereas cells medial to the eye formed the lateral ethmoid plate and trabeculae and a posterior population formed the mandible. miR-140 overexpression and estradiol inhibition of Hedgehog signaling resulted in cleft development, with failed migration of the anterior cell population to form the median ethmoid plate. The sox10:kaede transgenic line provides a useful tool for neural crest lineage analysis. These studies illustrate the advantages of the zebrafish model for application in morphogenetic studies of vertebrate craniofacial development.

Embryonic fate map of first pharyngeal arch structures in the sox10: kaede zebrafish transgenic model.

Lo sviluppo del palato nei vertebrati comporta la migrazione cresta neurale cranica, convergenza di prominenze facciali e l&#39;estensione del quadro cartilagineo. Disregolazione del palatogenesis risultati nella Schisi orofacciale, che rappresentano le pi&ugrave; comuni malformazioni strutturali. Analisi dettagliata di zebrafish palatogenesis rivelato distinti meccanismi di morfogenesi palatale: estensione, proliferazione e integrazione. Mostriamo che wnt9a &egrave; richiesto per estensione palatale, in cui i condrociti formano un fronte proliferativo, subiscono cambiamenti morfologici e intercalano per formare la piastra etmoide. Nel frattempo, irf6 &egrave; richiesta specificamente per l&#39;integrazione di prominenze facciali lungo una cucitura a forma di V. Questo lavoro presenta un&#39;analisi meccanicistica della morfogenesi del palato in un contesto clinicamente rilevante.

Requisiti diversi per wnt9a e irf6 nei meccanismi di integrazione e di estensione durante la morfogenesi palato zebrafish.

&#x5728;&#x810A;&#x690E;&#x52A8;&#x7269;&#x4E2D;&#x53E3;&#x611F;&#x7684;&#x53D1;&#x5C55;&#x6D89;&#x53CA;&#x5230;&#x9885;&#x795E;&#x7ECF;&#x5D74;&#x8FC1;&#x79FB;&#xFF0C;&#x9762;&#x90E8;&#x7684;&#x65E5;&#x73E5;&#x7684;&#x6536;&#x655B;&#x6027;&#x548C;&#x8F6F;&#x9AA8;&#x6846;&#x67B6;&#x7684;&#x5EF6;&#x957F;&#x3002;Palatogenesis &#x5931;&#x8C03;&#x5BFC;&#x81F4;&#x5634;&#x5507;&#x53E3;&#x9762;&#x90E8;&#x7578;&#x5F62;&#xFF0C;&#x8FD9;&#x662F;&#x6700;&#x5E38;&#x89C1;&#x7684;&#x7ED3;&#x6784;&#x51FA;&#x751F;&#x7F3A;&#x9677;&#x3002;&#x6591;&#x9A6C;&#x9C7C; palatogenesis &#x7684;&#x8BE6;&#x7EC6;&#x7684;&#x5206;&#x6790;&#x63ED;&#x793A;&#x4E0D;&#x540C;&#x673A;&#x5236;&#x7684;&#x816D;&#x5F62;&#x6001;&#x53D1;&#x751F;&#xFF1A; &#x6269;&#x5C55;&#x3001; &#x6269;&#x6563;&#x548C;&#x878D;&#x5408;&#x3002;&#x6211;&#x4EEC;&#x663E;&#x793A;&#x8BE5; wnt9a &#x662F;&#x6240;&#x5FC5;&#x9700;&#x7684;&#x816D;&#x6269;&#x5C55;&#xFF0C;&#x5176;&#x4E2D;&#x8F6F;&#x9AA8;&#x7EC6;&#x80DE;&#x5F62;&#x6210;&#x589E;&#x751F;&#x6027;&#x9635;&#x7EBF;&#x3001; &#x5F62;&#x6001;&#x5B66;&#x6539;&#x53D8;&#x548C; intercalate&#xFF0C;&#x5F62;&#x6210;&#x7B5B;&#x7684;&#x677F;&#x3002;&#x540C;&#x65F6;&#xFF0C;&#x9700;&#x8981;&#x4E13;&#x95E8;&#x4E3A;&#x4E00;&#x4F53;&#x5316;&#x7684; V &#x5F62;&#x710A;&#x7F1D;&#x6CBF;&#x9762;&#x90E8;&#x73E5; irf6&#x3002;&#x8FD9;&#x9879;&#x5DE5;&#x4F5C;&#x5728;&#x4E34;&#x5E8A;&#x4E0A;&#x6709;&#x5173;&#x65B9;&#x9762;&#x4ECB;&#x7ECD;&#x4E86;&#x816D;&#x90E8;&#x5F62;&#x6001;&#x53D1;&#x751F;&#x673A;&#x7406;&#x5206;&#x6790;&#x3002;

Wnt9a &#x548C; irf6 &#x5728;&#x6591;&#x9A6C;&#x9C7C;&#x7684;&#x5473;&#x89C9;&#x5F62;&#x6001;&#x53D1;&#x751F;&#x671F;&#x95F4;&#x7684;&#x6269;&#x5C55;&#x548C;&#x96C6;&#x6210;&#x673A;&#x5236;&#x7684;&#x4E0D;&#x540C;&#x8981;&#x6C42;&#x3002;

Development of the palate in vertebrates involves cranial neural crest migration, convergence of facial prominences and extension of the cartilaginous framework. Dysregulation of palatogenesis results in orofacial clefts, which represent the most common structural birth defects. Detailed analysis of zebrafish palatogenesis revealed distinct mechanisms of palatal morphogenesis: extension, proliferation and integration. We show that wnt9a is required for palatal extension, wherein the chondrocytes form a proliferative front, undergo morphological change and intercalate to form the ethmoid plate. Meanwhile, irf6 is required specifically for integration of facial prominences along a V-shaped seam. This work presents a mechanistic analysis of palate morphogenesis in a clinically relevant context.

Distinct requirements for wnt9a and irf6 in extension and integration mechanisms during zebrafish palate morphogenesis.

D&eacute;veloppement du Palais chez les vert&eacute;br&eacute;s implique la migration de cr&ecirc;te neurale cr&acirc;nienne, convergence des protub&eacute;rances osseuses du visage et l&#39;extension du cadre cartilagineux. Dysregulation de palatogenesis se traduit par des fentes oro-faciale, qui repr&eacute;sentent des d&eacute;fauts de naissance structurels les plus communs. Une analyse d&eacute;taill&eacute;e du poisson-z&egrave;bre palatogenesis a r&eacute;v&eacute;l&eacute; des m&eacute;canismes distincts de la morphog&eacute;n&egrave;se palatale : extension, de prolif&eacute;ration et d&#39;int&eacute;gration. Nous montrons que wnt9a est n&eacute;cessaire pour l&#39;extension palatine, dans lequel les chondrocytes forment un front prolif&eacute;ratif, subissent des modifications morphologiques et intercaler pour former la plaque de l&#39;ethmo&iuml;de. Dans le m&ecirc;me temps, il faut irf6 sp&eacute;cifiquement pour l&#39;int&eacute;gration des protub&eacute;rances du visage le long d&#39;une couture en forme de V. Cet ouvrage pr&eacute;sente une analyse m&eacute;caniste de la morphog&eacute;n&egrave;se de la bouche dans un contexte clinique pertinent.

Exigences distinctes pour les wnt9a et irf6 en extension et int&eacute;gration des m&eacute;canismes au cours de la morphogen&egrave;se de palais de poisson-z&egrave;bre.

Entwicklung des Gaumens bei Wirbeltieren geh&ouml;ren kraniale Neuralleiste Migration, Konvergenz der Gesichts Vorspr&uuml;ngen und Erweiterung der knorpelige Rahmen. Dysregulation der Palatogenesis f&uuml;hrt orofazialen Spalten, die die h&auml;ufigste strukturelle Geburtsfehler darstellen. Detaillierte Analyse der Zebrafisch Palatogenesis offenbart verschiedene Mechanismen der palatalen Morphogenese: Erweiterung sowie Verbreitung und Integration. Wir zeigen, dass wnt9a f&uuml;r palataler Erweiterung erforderlich ist, worin die Chondrozyten eine proliferative Front, morphologische Ver&auml;nderungen zu unterziehen und einzulagern um die ethmoid Platte zu bilden. Inzwischen ist irf6 speziell f&uuml;r die Integration des Gesichts Vorspr&uuml;ngen entlang eine v-f&ouml;rmige Naht erforderlich. Dieses Werk stellt eine mechanistische Analyse der Gaumen Morphogenese in einem klinisch relevanten Kontext.

Unterschiedliche Anforderungen f&uuml;r wnt9a und irf6 in Erweiterung und Integration Mechanismen w&auml;hrend Zebrafisch Gaumen Morphogenese.

&#x958B;&#x767A;&#x53E3;&#x84CB;&#x810A;&#x690E;&#x52D5;&#x7269;&#x306E;&#x982D;&#x84CB;&#x795E;&#x7D4C;&#x5824;&#x79FB;&#x884C;&#x3001;&#x9854;&#x9762;&#x30D7;&#x30ED;&#x30DF;&#x30CD;&#x30F3;&#x30B9;&#x306E;&#x53CE;&#x675F;&#x3068;&#x8EDF;&#x9AA8;&#x30D5;&#x30EC;&#x30FC;&#x30E0;&#x30EF;&#x30FC;&#x30AF;&#x306E;&#x62E1;&#x5F35;&#x3057;&#x307E;&#x3059;&#x3002;&#x305D;&#x306E;&#x5236;&#x5FA1;&#x7834;&#x7DBB;&#x4E8C;&#x6700;&#x3082;&#x4E00;&#x822C;&#x7684;&#x306A;&#x69CB;&#x9020;&#x306E;&#x51FA;&#x751F;&#x6642;&#x6B20;&#x640D;&#x3092;&#x8868;&#x3059;&#x53E3;&#x8154;&#x9854;&#x9762;&#x88C2;&#x306E;&#x7D50;&#x679C;&#x3057;&#x307E;&#x3059;&#x3002;&#x30BC;&#x30D6;&#x30E9;&#x30D5;&#x30A3;&#x30C3;&#x30B7;&#x30E5;&#x4E8C;&#x306E;&#x8A73;&#x7D30;&#x306A;&#x5206;&#x6790;&#x306F;&#x53E3;&#x84CB;&#x306E;&#x5F62;&#x614B;&#x5F62;&#x6210;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x305F;: &#x62E1;&#x5F35;&#x5B50;&#x3001;&#x5897;&#x6B96;&#x3068;&#x7D71;&#x5408;&#x3002;&#x305D;&#x306E; wnt9a &#x306F;&#x53E3;&#x84CB;&#x306E;&#x62E1;&#x5F35;&#x5B50;&#x306B;&#x3064;&#x3044;&#x3066;&#x306F;&#x3001;&#x524D;&#x8A18;&#x306E;&#x8EDF;&#x9AA8;&#x7D30;&#x80DE;&#x5897;&#x6B96;&#x6226;&#x7DDA;&#x306E;&#x5F62;&#x6210;&#x5F62;&#x614B;&#x5909;&#x5316;&#x3092;&#x53D7;&#x3051;&#x308B;&#x3057;&#x3001;&#x7BE9;&#x9AA8;&#x677F;&#x3092;&#x5F62;&#x6210;&#x3059;&#x308B;&#x30A4;&#x30F3;&#x30BF;&#x30FC;&#x30AB; &#x30EC;&#x30FC;&#x30C8;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3059;.&#x4E00;&#x65B9;&#x3001;irf6 &#x306F;&#x3001;&#x5177;&#x4F53;&#x7684;&#x306B;&#x306F; V &#x5B57;&#x306E;&#x7E2B;&#x3044;&#x76EE;&#x306B;&#x6CBF;&#x3063;&#x3066;&#x9854;&#x30D7;&#x30ED;&#x30DF;&#x30CD;&#x30F3;&#x30B9;&#x306E;&#x7D71;&#x5408;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3059;&#x3002;&#x3053;&#x306E;&#x4ED5;&#x4E8B;&#x53E3;&#x84CB;&#x306E;&#x5F62;&#x614B;&#x5F62;&#x6210;&#x306E;&#x6A5F;&#x69CB;&#x7684;&#x89E3;&#x6790;&#x3067;&#x81E8;&#x5E8A;&#x7684;&#x306B;&#x95A2;&#x9023;&#x3059;&#x308B;&#x30B3;&#x30F3;&#x30C6;&#x30AD;&#x30B9;&#x30C8;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;

&#x660E;&#x78BA;&#x306A;&#x8981;&#x4EF6;&#x3092; wnt9a &#x3068; irf6 &#x30BC;&#x30D6;&#x30E9;&#x30D5;&#x30A3;&#x30C3;&#x30B7;&#x30E5;&#x306E;&#x53E3;&#x84CB;&#x306E;&#x5F62;&#x614B;&#x5F62;&#x6210;&#x306B;&#x304A;&#x3051;&#x308B;&#x62E1;&#x5F35;&#x3068;&#x7D71;&#x5408;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3067;&#x3002;

&#xCC99;&#xCD94; &#xB3D9;&#xBB3C;&#xC5D0;&#xC11C; &#xBBF8;&#xAC01;&#xC758; &#xAC1C;&#xBC1C; &#xB450;&#xAC1C;&#xACE8; &#xC2E0;&#xACBD; &#xD06C;&#xB808;&#xC2A4;&#xD2B8; &#xB9C8;&#xC774;&#xADF8;&#xB808;&#xC774;&#xC158;&#xC744;, &#xC5BC;&#xAD74; &#xAD74;&#xC9C0;&#xC758; &#xC735;&#xD569; &#xBC0F; &#xC5F0;&#xACE8; &#xD504;&#xB808;&#xC784; &#xC6CC;&#xD06C;&#xC758; &#xD655;&#xC7A5;&#xC744; &#xD3EC;&#xD568; &#xD55C;&#xB2E4;. &#xAC00;&#xC7A5; &#xC77C;&#xBC18;&#xC801;&#xC778; &#xAD6C;&#xC870;&#xC801; &#xAE30;&#xD615; &#xB300;&#xD45C; orofacial clefts Dysregulation palatogenesis&#xC758; &#xACB0;&#xACFC;. Zebrafish palatogenesis&#xC758; &#xC0C1;&#xC138;&#xD55C; &#xBD84;&#xC11D; &#xACF5;&#xAC1C; &#xACBD;&#xAD6C;&#xAC1C; morphogenesis&#xC758; &#xB3C5;&#xD2B9;&#xD55C; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;: &#xD655;&#xC7A5;, &#xD655;&#xC0B0; &#xBC0F; &#xD1B5;&#xD569;. &#xC6B0;&#xB9AC;&#xB294; &#xADF8; wnt9a&#xB294; &#xACBD;&#xAD6C;&#xAC1C; &#xD655;&#xC7A5;, &#xC5B4;&#xB5A4; &#xC810;&#xC5D0;&#xC11C; chondrocytes &#xC99D;&#xC2DD; &#xD504;&#xB7F0;&#xD2B8;&#xB97C; &#xD615;&#xC131;, &#xD615;&#xD0DC;&#xD559; &#xBCC0;&#xD654; &#xBC1B;&#xC544;&#xC57C;&#xC640; &#xC0AC; &#xACE8; &#xD310; &#xD615;&#xC131; intercalate &#xD544;&#xC694; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. &#xD55C;&#xD3B8;, irf6&#xB294; V &#xC790;&#xD615; &#xC194; &#xAE30;&#xB97C; &#xB530;&#xB77C; &#xC5BC;&#xAD74; &#xAD74;&#xC9C0;&#xC758; &#xD1B5;&#xD569;&#xC744; &#xC704;&#xD574; &#xD2B9;&#xBCC4;&#xD788; &#xD544;&#xC694; &#xD569;&#xB2C8;&#xB2E4;. &#xC774; &#xC791;&#xD488; &#xC784;&#xC0C1; &#xAD00;&#xB828; &#xCEE8;&#xD14D;&#xC2A4;&#xD2B8;&#xC5D0;&#xC11C; &#xAD6C;&#xAC1C; morphogenesis&#xC758; &#xAE30;&#xACC4; &#xB860; &#xC801;&#xC778; &#xBD84;&#xC11D;&#xC744; &#xC81C;&#xACF5;&#xD569;&#xB2C8;&#xB2E4;.

Wnt9a &#xBC0F; irf6 zebrafish &#xAD6C;&#xAC1C; morphogenesis &#xB3D9;&#xC548; &#xD655;&#xC7A5; &#xBC0F; &#xD1B5;&#xD569; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC5D0; &#xB300; &#xD55C; &#xAC1C;&#xBCC4; &#xC694;&#xAD6C; &#xC0AC;&#xD56D;.

Desarrollo del paladar en los vertebrados implica la migraci&oacute;n de la cresta neural craneal, convergencia de prominencias faciales y extensi&oacute;n del marco cartilaginoso. Dysregulation de palatogenesis da lugar a hendiduras orofaciales, que representan los defectos de nacimiento estructurales m&aacute;s comunes. An&aacute;lisis detallado de pez cebra palatogenesis revel&oacute; distintos mecanismos de morfog&eacute;nesis palatal: extensi&oacute;n, la proliferaci&oacute;n y la integraci&oacute;n. Mostramos que wnt9a se requiere para la extensi&oacute;n palatal, en donde los condrocitos formar un frente proliferativo, someterse a cambios morfol&oacute;gicos e intercalar para formar la placa etmoide. Mientras tanto, irf6 se requiere espec&iacute;ficamente para integraci&oacute;n de prominencias faciales a lo largo de una costura en forma de V. Este trabajo presenta un an&aacute;lisis mecanicista de la morfog&eacute;nesis del paladar en un contexto cl&iacute;nico relevante.

Distintos requisitos para wnt9a y irf6 en mecanismos de integraci&oacute;n y extensi&oacute;n durante la morfog&eacute;nesis de boca de pez cebra.

Regulation of convergence and extension by wnt-frizzled signaling is a common theme in embryogenesis. This study examines the functional requirements of frzb and fzd7a in convergence and extension mechanisms during craniofacial development. Using a morpholino knockdown approach, we found that frzb and fzd7a are dispensable for directed migration of the bilateral trabeculae, but necessary for the convergence and extension of the palatal elements, where the extension process is mediated by chondrocyte proliferation, morphologic change and intercalation. In contrast, frzb and fzd7a are required for convergence of the mandibular prominences, where knockdown of either frzb or fzd7a resulted in complete loss of lower jaw structures. Further, we found that bapx1 was specifically downregulated in the wnt9a/frzb/fzd7a morphants, while general neural crest markers were unaffected. In addition, expression of wnt9a and frzb was also absent in the edn-/- mutant. Notably, over-expression of bapx1 was sufficient to partially rescue mandibular elements in the wnt9a/frzb/fzd7a morphants, demonstrating genetic epistasis of bapx1 acting downstream of edn1 and wnt9a/frzb/fzd7a in lower jaw development. This study underscores the important role of wnt-frizzled signaling in convergence and extension in palate and craniofacial morphogenesis, distinct regulation of upper vs. lower jaw structures, and integration of wnt-frizzled with endothelin signaling to coordinate shaping of the facial form.

Article	Year
Embryonic fate map of first pharyngeal arch structures in the sox10: kaede zebrafish transgenic model. The Journal of craniofacial surgery\| PubMed ID: 22948622	2012
Distinct requirements for wnt9a and irf6 in extension and integration mechanisms during zebrafish palate morphogenesis. Development (Cambridge, England)\| PubMed ID: 23154410	2012
Requirement for frzb and fzd7a in cranial neural crest convergence and extension mechanisms during zebrafish palate and jaw morphogenesis. Developmental biology\| PubMed ID: 23806211	2013

Max Dougherty

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