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

Detta arbete har finansierats av R01-DE018234 och R01-DE019638.

1. Förbereda givare Tissue <ol><li> Förbered kulturmedia slipa glas stift, vässa volfram nålar. </li><li> Samla embryo från Shell, tvätta i iskallt PBS. <ol class="lalpha"><li> Använd en 10 ml spruta och en 18 gauge nål bort 1,0 ml av albumin från den spetsiga änden av äggskal. </li><li> Gör ett litet hål på toppen av skalet med den punkt sax och klipp sedan en rund öppning att exponera embryot </li></ol></li><li> Ta bort huvud och placera i DMEM (serum gratis, rumstemperatur) </li><li> Dissekera Fronto...

<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. 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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. 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Med denna transplantation metoden har vi kunnat fastställa att ektoderm innehåller signalering information som reglerar dorsoventral polaritet och proximodistal förlängning av överkäken. Likheten mellan resultat vid användning av vaktel eller musen ektoderm, och bevarandet av molekylära signaler i denna vävnad bland många arter 6,11 indikerar att detta är ett starkt konservativa signalering centrum bland ryggradsdjuren. Dessutom har andra forskare använt liknande tekniker för att testa signalerin...

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<td>1x PBS</td>
<td>&nbsp;</td>
<td>TEK</td>
<td>TEKZR114</td>
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<td>DMEM</td>
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<td>CCFDA003</td>
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<td>A7906</td>
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<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>
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<td>Scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14058-11</td>
<td>&nbsp;</td>
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<td>Spring Scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
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<td>Needle holder</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>26016-12</td>
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<td>Tungsten Needle</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>26000</td>
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<td>Microcapillary tube </td>
<td>&nbsp;</td>
<td>Drummond Scientific Company</td>
<td>3-000-225-G</td>
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<td>Pasteur Pipets</td>
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<td>Fisher</td>
<td>13-678-6B</td>
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<td>Spring scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>15010-11 </td>
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<td>Blade holder</td>
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<td>Fine Science Tools</td>
<td>10052-11 </td>
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<td>Razor blade</td>
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<td>Fine Science Tools</td>
<td>10050-00</td>
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assessing signaling properties of ectodermal epithelia during craniofacial development

Tillgängligheten av aviär embryon har hjälpt experimentell embryologer förstå öden celler under utveckling och roll vävnad interaktioner som reglerar mönstring och morfogenes av ryggradsdjur (t.ex. 1, 2, 3, 4). Här visar vi en metod som utnyttjar denna tillgänglighet för att testa signalering och egenskaper mönstring av ektodermal vävnader vid ansikts utveckling. I dessa försök skapar vi vaktel-chick 5 eller mus-chick 6 chimärer genom att transplantera ytan cefaliska ektoderm som täcker den övre käken från vaktel eller musen på antingen samma region eller en ektopisk region kycklingembryon. Användningen av vaktlar som givare vävnad för transplantation till kycklingar har utvecklats för att dra fördel av en nucleolar markör som finns i vaktlar men inte kycklingembryoceller, vilket gör utredare att skilja värd och vävnader donator 7. Likaså är ett återkommande inslag som finns i musens arvsmassa och uttrycks ubiquitously, som tillåter oss att skilja värd och vävnader givare mus-chick chimärer 8. Att använda musen ektoderm som givare vävnad kommer kraftigt att utöka vår förståelse för dessa vävnad interaktioner, eftersom detta ger oss möjlighet att testa signalering egenskaper ektoderm härleds från olika mutant embryon.

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

Den här artikeln beskriver en teknik vävnadstransplantation som var utformat för att testa signalsystemet och egenskaper mönstring av ytor cefaliska ektoderm under kraniofaciala utveckling.

Bedöma Signalering egenskaper Ectodermal epitel Under Kraniofaciala utveckling

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