Name: assessing signaling properties of ectodermal epithelia during craniofacial development
Uploaded: 2011-03-24T11:00:00
Description: Watch this Scientific Journal Video about Assessing Signaling Properties of Ectodermal Epithelia During Craniofacial Development at JoVE.com

Questo lavoro è stato finanziato dalla R01-R01-DE018234 e DE019638.

1. Preparazione del tessuto del donatore <ol><li> Preparare terreni di coltura, affinare le spine di vetro, affilare gli aghi di tungsteno. </li><li> Raccogliere embrione dal guscio, lavare in gelide PBS. <ol class="lalpha"><li> Usando una siringa da 10 ml e 18 gauge rimuovere 1,0 ml di albumina da l&#39;estremità appuntita del guscio d&#39;uovo. </li><li> Fai un piccolo foro sulla parte superiore del guscio utilizzando il punto di forbici, e poi tagliare un&#39;apertura circolare per esporre l&#39;embrione </li></ol...

<ol>
	<li>Noden, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Role+of+the+Neural+Crest+in+Patterning+of+Avian+Cranial+Skeletal,+Connective,+and+Muscle+Tissues.">The Role of the Neural Crest in Patterning of Avian Cranial Skeletal, Connective, and Muscle Tissues.</a> Developmental Biology. 96, 144-144 (1983).</li><li>Bronner-Fraser, M., Stern, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+Mesodermal+Tissues+on+Avian+Neural+Crest+Cell+Migration.">Effects of Mesodermal Tissues on Avian Neural Crest Cell Migration.</a> Developmental Biology. 143, 213-213 (1991).</li><li>Schneider, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+crest+can+form+cartilages+normally+derived+from+mesoderm+during+development+of+the+avian+head+skeleton.">Neural crest can form cartilages normally derived from mesoderm during development of the avian head skeleton.</a> Developmental Biology. 208, 441-441 (1999).</li><li>Couly, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interactions+between+Hox-negative+cephalic+neural+crest+cells+and+the+foregut+endoderm+in+patterning+the+facial+skeleton+in+the+vertebrate+head.">Interactions between Hox-negative cephalic neural crest cells and the foregut endoderm in patterning the facial skeleton in the vertebrate head.</a> Development. 129, 1061-1061 (2002).</li><li>Evans, D. J., Noden, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+relations+between+avian+craniofacial+neural+crest+and+paraxial+mesoderm+cells.">Spatial relations between avian craniofacial neural crest and paraxial mesoderm cells.</a> Dev Dyn. , (2006).</li><li>Hu, D., Marcucio, R., Helms, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+zone+of+frontonasal+ectoderm+regulates+patterning+and+growth+in+the+face.">A zone of frontonasal ectoderm regulates patterning and growth in the face.</a> Development. 130, 1749-1749 (2003).</li><li>Hu, D., Marcucio, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unique+organization+of+the+frontonasal+ectodermal+zone+in+birds+and+mammals.">Unique organization of the frontonasal ectodermal zone in birds and mammals.</a> Dev Biol. 325, 200-200 (2009).</li><li>Le Li&#232;vre, C. S., Le Douarin, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesenchymal+derivatives+of+the+neural+crest:+analysis+of+chimaeric+quail+and+chick+embryos.">Mesenchymal derivatives of the neural crest: analysis of chimaeric quail and chick embryos.</a> Journal of Embryology and Experimental Morphology. 34, 125-125 (1975).</li><li>Bollag, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+a+repetitive+mouse+B2+element+to+identify+transplanted+mouse+cells+in+mouse-chick+chimeras.">Use of a repetitive mouse B2 element to identify transplanted mouse cells in mouse-chick chimeras.</a> Exp Cell Res. 248, 75-75 (1999).</li><li>Korn, M. J., Cramer, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Windowing+chicken+eggs+for+developmental+studies.">Windowing chicken eggs for developmental studies.</a> J Vis Exp. , (2007).</li><li>Eames, B. F., Schneider, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quail-duck+chimeras+reveal+spatiotemporal+plasticity+in+molecular+and+histogenic+programs+of+cranial+feather+development.">Quail-duck chimeras reveal spatiotemporal plasticity in molecular and histogenic programs of cranial feather development.</a> Development. 132, 1499-1499 (2005).</li><li>Odent, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+the+Sonic+hedgehog+(SHH+)+gene+during+early+human+development+and+phenotypic+expression+of+new+mutations+causing+holoprosencephaly.">Expression of the Sonic hedgehog (SHH ) gene during early human development and phenotypic expression of new mutations causing holoprosencephaly.</a> Hum Mol Genet. 8, 1683-1683 (1999).</li><li>Szabo-Rogers, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+skeletogenic+patterning+roles+for+the+olfactory+pit.">Novel skeletogenic patterning roles for the olfactory pit.</a> Development. 136, 219-219 (2009).</li></ol>

Usando questo metodo di trapianto ha permesso di determinare che l&#39;ectoderma contiene informazioni di segnalazione che regola la polarità dorso-ventrale e l&#39;estensione prossimo-distale della mascella superiore. La somiglianza dei risultati quando si usa quaglia o ectoderma mouse e la conservazione dei segnali molecolari in questo tessuto tra le molte specie 6,11 indica che questo è un centro altamente conservato di segnalazione tra i vertebrati. Inoltre, altri ricercatori hanno usato tecniche simili...

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		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>1x PBS</td>
<td>&nbsp;</td>
<td>TEK</td>
<td>TEKZR114</td>
<td>&nbsp;</td>
<tr>
<td>DMEM</td>
<td>&nbsp;</td>
<td>UCSF</td>
<td>CCFDA003</td>
<td>&nbsp;</td>
<tr>
<td>BSA</td>
<td>&nbsp;</td>
<td>SIGMA</td>
<td>A7906</td>
<td>&nbsp;</td>
<tr>
<td>Dispase</td>
<td>&nbsp;</td>
<td>GIBCO</td>
<td>17105-041</td>
<td>&nbsp;</td>
<tr>
<td>35x10 mm Petri dish</td>
<td>&nbsp;</td>
<td>Falcon</td>
<td>1008</td>
<td>&nbsp;</td>
<tr>
<td>No. 5 Dumont forceps</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11252-20</td>
<td>&nbsp;</td>
<tr>
<td>Scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14058-11</td>
<td>&nbsp;</td>
<tr>
<td>Spring Scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>15010-11</td>
<td>&nbsp;</td>
<tr>
<td>Needle holder</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>26016-12</td>
<td>&nbsp;</td>
<tr>
<td>Tungsten Needle</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>26000</td>
<td>&nbsp;</td>
<tr>
<td>Microcapillary tube </td>
<td>&nbsp;</td>
<td>Drummond Scientific Company</td>
<td>3-000-225-G</td>
<td>&nbsp;</td>
<tr>
<td>Pasteur Pipets</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>13-678-6B</td>
<td>&nbsp;</td>
<tr>
<td>Spring scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>15010-11 </td>
<td>&nbsp;</td>
<tr>
<td>Blade holder</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>10052-11 </td>
<td>&nbsp;</td>
<tr>
<td>Razor blade</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>10050-00</td>
<td>&nbsp;</td>		</tbody>
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assessing signaling properties of ectodermal epithelia during craniofacial development

L&#39;accessibilità degli embrioni aviaria ha aiutato embriologi sperimentali capire il destino delle cellule durante lo sviluppo e il ruolo delle interazioni tessuti che regolano patterning e la morfogenesi dei vertebrati (ad esempio, 1, 2, 3, 4). Qui, illustrare un metodo che sfrutta questa accessibilità per testare la segnalazione e le proprietà patterning dei tessuti durante lo sviluppo ectodermica del viso. In questi esperimenti, creiamo quaglia-chick 5 o topo-chick 6 chimere trapiantando l&#39;ectoderma cefalico superficie che copre la mascella superiore da quaglia o mouse sulla entrambi della stessa regione o di una regione ectopica di embrioni di pollo. L&#39;uso di quaglia come tessuto del donatore per il trapianto in pulcini è stato sviluppato per trarre vantaggio da un marker nucleolare presente in quaglie, ma non le cellule pulcino, permettendo così agli investigatori di distinguere host e tessuti del donatore 7. Allo stesso modo, un elemento ripetitivo è presente nel genoma del topo ed è espressa ubiquitariamente, che ci permette di distinguere host e tessuti del donatore nel topo-chick chimere 8. L&#39;uso di ectoderma mouse come tessuto del donatore amplieranno enormemente la nostra comprensione di queste interazioni dei tessuti, perché questo ci permetterà di testare le proprietà di segnalazione di ectoderma derivate da embrioni mutanti vari.

Watch this Scientific Journal Video about Assessing Signaling Properties of Ectodermal Epithelia During Craniofacial Development at JoVE.com

Assessing Signaling Properties of Ectodermal Epithelia During Craniofacial Development

Questo articolo descrive una tecnica di trapianto di tessuto che è stato progettato per verificare la segnalazione e le proprietà di patterning ectoderma cefalico superficie durante lo sviluppo cranio-facciale.

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