Name: the power of simplicity: sea urchin embryos as in vivo developmental models for studying complex cell-to-cell signaling network interactions
Uploaded: 2017-02-16T14:00:00
Description: Watch this Scientific Journal Video about The Power of Simplicity: Sea Urchin Embryos as in Vivo Developmental Models for Studying Complex Cell-to-cell Signaling Network Interactions at JoVE.com

We would like to thank Dr. Robert Angerer for his careful reading and editing of the manuscript. NIH R15HD088272-01 as well as the Office of Research and Development, and Department of Biological Sciences at Mississippi State University provided support for this project to RCR.

1. Hoher Durchsatz Morpholino Design Strategy <ol><li> Identifizieren eines Gens (e) von Interesse (zB Kandidatengen - Ansatz, cis-regulatorische Analyse, RNA-Seq und / oder Proteom - Differential - Bildschirme). </li><li> Verwenden Sie genomischen, Transkriptom und Genexpressionsdaten auf häufig aktualisierte Webseiten (zB SpBase http://www.echinobase.org 20 und S. purpuratus Genom suchen http: ///urchin.nidcr.nih.gov/blast/index .html), um festzustellen, dass der R...

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	<li>Erwin, D. H., Davidson, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+evolution+of+hierarchical+gene+regulatory+networks.">The evolution of hierarchical gene regulatory networks.</a> Nature reviews. Genetics. 10, 141-148 (2009).</li><li>Peter, I. S., Davidson, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolution+of+gene+regulatory+networks+controlling+body+plan+development.">Evolution of gene regulatory networks controlling body plan development.</a> Cell. 144, 970-985 (2011).</li><li>Borggrefe, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Notch+intracellular+domain+integrates+signals+from+Wnt,+Hedgehog,+TGFbeta/BMP+and+hypoxia+pathways.">The Notch intracellular domain integrates signals from Wnt, Hedgehog, TGFbeta/BMP and hypoxia pathways.</a> Biochimica et biophysica acta. 1863, 303-313 (2016).</li><li>Range, R. C., Angerer, R. C., Angerer, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integration+of+canonical+and+noncanonical+Wnt+signaling+pathways+patterns+the+neuroectoderm+along+the+anterior-posterior+axis+of+sea+urchin+embryos.">Integration of canonical and noncanonical Wnt signaling pathways patterns the neuroectoderm along the anterior-posterior axis of sea urchin embryos.</a> PLoS Biol. 11, e1001467 (2013).</li><li>Cleary, M. A., van Osch, G. J., Brama, P. A., Hellingman, C. A., Narcisi, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FGF,+TGFbeta+and+Wnt+crosstalk:+embryonic+to+in+vitro+cartilage+development+from+mesenchymal+stem+cells.">FGF, TGFbeta and Wnt crosstalk: embryonic to in vitro cartilage development from mesenchymal stem cells.</a> Journal of tissue engineering and regenerative medicine. 9, 332-342 (2015).</li><li>Lapraz, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RTK+and+TGF-beta+signaling+pathways+genes+in+the+sea+urchin+genome.">RTK and TGF-beta signaling pathways genes in the sea urchin genome.</a> Dev Biol. 300, 132-152 (2006).</li><li>Pires-daSilva, A., Sommer, 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=The+evolution+of+signalling+pathways+in+animal+development.">The evolution of signalling pathways in animal development.</a> Nature reviews. Genetics. 4, 39-49 (2003).</li><li>Sethi, A. J., Wikramanayake, R. M., Angerer, R. C., Range, R. C., Angerer, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sequential+signaling+crosstalk+regulates+endomesoderm+segregation+in+sea+urchin+embryos.">Sequential signaling crosstalk regulates endomesoderm segregation in sea urchin embryos.</a> Science. 335, 590-593 (2012).</li><li>Range, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Specification+and+positioning+of+the+anterior+neuroectoderm+in+deuterostome+embryos.">Specification and positioning of the anterior neuroectoderm in deuterostome embryos.</a> Genesis. 52, 222-234 (2014).</li><li>Petersen, C. P., Reddien, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wnt+signaling+and+the+polarity+of+the+primary+body+axis.">Wnt signaling and the polarity of the primary body axis.</a> Cell. 139, 1056-1068 (2009).</li><li>Lapraz, F., Haillot, E., Lepage, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+deuterostome+origin+of+the+Spemann+organiser+suggested+by+Nodal+and+ADMPs+functions+in+Echinoderms.">A deuterostome origin of the Spemann organiser suggested by Nodal and ADMPs functions in Echinoderms.</a> Nature communications. 6, 8434 (2015).</li><li>Kikuchi, A., Yamamoto, H., Sato, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+activation+mechanisms+of+Wnt+signaling+pathways.">Selective activation mechanisms of Wnt signaling pathways.</a> Trends in cell biology. 19, 119-129 (2009).</li><li>Hogan, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+morphogenetic+proteins:+multifunctional+regulators+of+vertebrate+development.">Bone morphogenetic proteins: multifunctional regulators of vertebrate development.</a> Genes Dev. 10, 1580-1594 (1996).</li><li>Houart, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+the+telencephalon+during+gastrulation+by+local+antagonism+of+Wnt+signaling.">Establishment of the telencephalon during gastrulation by local antagonism of Wnt signaling.</a> Neuron. 35, 255-265 (2002).</li><li>Bertrand, S., Escriva, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolutionary+crossroads+in+developmental+biology:+amphioxus.">Evolutionary crossroads in developmental biology: amphioxus.</a> Development. 138, 4819-4830 (2011).</li><li>Rottinger, E., Lowe, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolutionary+crossroads+in+developmental+biology:+hemichordates.">Evolutionary crossroads in developmental biology: hemichordates.</a> Development. 139, 2463-2475 (2012).</li><li>Genome Sequencing Sea Urchin, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genome+of+the+sea+urchin+Strongylocentrotus+purpuratus.">The genome of the sea urchin Strongylocentrotus purpuratus.</a> Science. 314, 941-952 (2006).</li><li>Ben-Tabou de-Leon, S., Su, Y. H., Lin, K. T., Li, E., Davidson, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+regulatory+control+in+the+sea+urchin+aboral+ectoderm:+spatial+initiation,+signaling+inputs,+and+cell+fate+lockdown.">Gene regulatory control in the sea urchin aboral ectoderm: spatial initiation, signaling inputs, and cell fate lockdown.</a> Dev Biol. 374, 245-254 (2013).</li><li>Saudemont, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ancestral+regulatory+circuits+governing+ectoderm+patterning+downstream+of+Nodal+and+BMP2/4+revealed+by+gene+regulatory+network+analysis+in+an+echinoderm.">Ancestral regulatory circuits governing ectoderm patterning downstream of Nodal and BMP2/4 revealed by gene regulatory network analysis in an echinoderm.</a> PLoS Genet. 6, e1001259 (2010).</li><li>Cameron, R. A., Samanta, M., Yuan, A., He, D., Davidson, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SpBase:+the+sea+urchin+genome+database+and+web+site.">SpBase: the sea urchin genome database and web site.</a> Nucleic Acids Res. 37, D750-D754 (2009).</li><li>Stepicheva, N. A., Song, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+throughput+microinjections+of+sea+urchin+zygotes.">High throughput microinjections of sea urchin zygotes.</a> Journal of visualized experiments : JoVE. , e50841 (2014).</li><li>Cheers, M. S., Ettensohn, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+microinjection+of+fertilized+eggs.">Rapid microinjection of fertilized eggs.</a> Methods in cell biology. 74, 287-310 (2004).</li><li>Arenas-Mena, C., Cameron, A. R., Davidson, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+expression+of+Hox+cluster+genes+in+the+ontogeny+of+a+sea+urchin.">Spatial expression of Hox cluster genes in the ontogeny of a sea urchin.</a> Development. , 4631-4643 (2000).</li><li>Sethi, A. J., Angerer, R. C., Angerer, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multicolor+labeling+in+developmental+gene+regulatory+network+analysis.">Multicolor labeling in developmental gene regulatory network analysis.</a> Methods in molecular biology. , 249-262 (2014).</li><li>Wikramanayake, A. H., Huang, L., Klein, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=beta-Catenin+is+essential+for+patterning+the+maternally+specified+animal-vegetal+axis+in+the+sea+urchin+embryo.">beta-Catenin is essential for patterning the maternally specified animal-vegetal axis in the sea urchin embryo.</a> Proc Natl Acad Sci U S A. 95, 9343 (1998).</li><li>Yaguchi, S., Yaguchi, J., Angerer, R. C., Angerer, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Wnt-FoxQ2-nodal+pathway+links+primary+and+secondary+axis+specification+in+sea+urchin+embryos.">A Wnt-FoxQ2-nodal pathway links primary and secondary axis specification in sea urchin embryos.</a> Dev Cell. 14, 97-107 (2008).</li><li>Molina, M. D., de Croze, N., Haillot, E., Lepage, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nodal:+master+and+commander+of+the+dorsal-ventral+and+left-right+axes+in+the+sea+urchin+embryo.">Nodal: master and commander of the dorsal-ventral and left-right axes in the sea urchin embryo.</a> Curr Opin Genet Dev. 23, 445-453 (2013).</li><li>Range, R. C., Glenn, T. D., Miranda, E., McClay, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=LvNumb+works+synergistically+with+Notch+signaling+to+specify+non-skeletal+mesoderm+cells+in+the+sea+urchin+embryo.">LvNumb works synergistically with Notch signaling to specify non-skeletal mesoderm cells in the sea urchin embryo.</a> Development. 135, 2445-2454 (2008).</li><li>Range, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cis-regulatory+analysis+of+nodal+and+maternal+control+of+dorsal-ventral+axis+formation+by+Univin,+a+TGF-beta+related+to+Vg1.">Cis-regulatory analysis of nodal and maternal control of dorsal-ventral axis formation by Univin, a TGF-beta related to Vg1.</a> Development. 134, 3649-3664 (2007).</li><li>Warner, J. F., Miranda, E. L., McClay, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+hedgehog+signaling+to+the+establishment+of+left-right+asymmetry+in+the+sea+urchin.">Contribution of hedgehog signaling to the establishment of left-right asymmetry in the sea urchin.</a> Dev Biol. 411, 314-324 (2016).</li><li>Rottinger, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FGF+signals+guide+migration+of+mesenchymal+cells,+control+skeletal+morphogenesis+[corrected]+and+regulate+gastrulation+during+sea+urchin+development.">FGF signals guide migration of mesenchymal cells, control skeletal morphogenesis [corrected] and regulate gastrulation during sea urchin development.</a> Development. 135, 353-365 (2008).</li><li>Warner, J. F., McCarthy, A. M., Morris, R. L., McClay, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hedgehog+signaling+requires+motile+cilia+in+the+sea+urchin.">Hedgehog signaling requires motile cilia in the sea urchin.</a> Mol Biol Evol. 31, 18-22 (2014).</li><li>Technau, U., Steele, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolutionary+crossroads+in+developmental+biology.">Evolutionary crossroads in developmental biology.</a> Cnidaria. Development. 138, 1447-1458 (2011).</li><li>Yaguchi, J., Takeda, N., Inaba, K., Yaguchi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cooperative+Wnt-Nodal+Signals+Regulate+the+Patterning+of+Anterior+Neuroectoderm.">Cooperative Wnt-Nodal Signals Regulate the Patterning of Anterior Neuroectoderm.</a> PLoS Genet. 12, e1006001 (2016).</li><li>Duboc, V., Rottinger, E., Besnardeau, L., Lepage, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nodal+and+BMP2/4+signaling+organizes+the+oral-aboral+axis+of+the+sea+urchin+embryo.">Nodal and BMP2/4 signaling organizes the oral-aboral axis of the sea urchin embryo.</a> Dev Cell. 6, 397-410 (2004).</li><li>Bradham, C. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chordin+is+required+for+neural+but+not+axial+development+in+sea+urchin+embryos.">Chordin is required for neural but not axial development in sea urchin embryos.</a> Dev Biol. 328, 221-233 (2009).</li><li>Su, Y. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+regulatory+networks+for+ectoderm+specification+in+sea+urchin+embryos.">Gene regulatory networks for ectoderm specification in sea urchin embryos.</a> Biochimica et biophysica acta. 1789, 261-267 (2009).</li><li>Lin, C. Y., Su, Y. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genome+editing+in+sea+urchin+embryos+by+using+a+CRISPR/Cas9+system.">Genome editing in sea urchin embryos by using a CRISPR/Cas9 system.</a> Dev Biol. 409, 420-428 (2016).</li></ol>

Die Methodik präsentiert hier ist ein Beispiel, das die Macht der Verwendung von Embryonen mit weniger genomischen und morphologische Komplexität als Wirbeltiere zeigt die Signalübertragungswege und KNS Regierungs grundlegenden Entwicklungsmechanismen .. Viele Labors verwenden ähnliche Tests während der frühen Seeigel Entwicklung zu verstehen, die zu sezieren Signalisierung in anderen Zellschicksal Spezifikation Ereignisse beteiligt Wege (zB Notch, Igel, TGF - β und FGF - Signalisierung) <sup class="xref...

Gene regulatorische Netzwerke (KNS) und Signaltransduktionswege schaffen die räumliche und zeitliche Expression von Genen während der embryonalen Entwicklung, die verwendet werden, um die erwachsenen tierischen Körper Plan zu bauen. Zell-Zell-Signalwege sind wesentliche Bestandteile dieser regulatorischen Netzwerke, die Bereitstellung der Mittel, durch die Zellen in Verbindung stehen. Diese zellulären Interaktionen etablieren und die Expression von regulatorischen und Differenzierung Gene in und zwischen den verschiedenen Gebieten während der Embryonalentwicklung 1, 2 zu verfeinern. Wechselwirkungen zwischen sezerniert extrazellulären Modulatoren (Liganden, Antagonisten), Rezeptoren und Co-Rezeptoren steuern die Aktivitäten von Signaltransduktionswegen. Eine Auswahl von intrazellulären Molekülen wandelt diese Eingänge in veränderter Genexpression, Abteilung führt, und / oder die Form einer Zelle. Während viele der Schlüsselmoleküle bei den extrazellulären und intrazellulären Spiegel in den Hauptwegen verwendet werden,bekannt ist, ist es eine unvollständige Kenntnis der Komplexität der einzelnen Signalwege zu einem großen Teil. Zusätzlich interagieren verschiedene Signalwege oft miteinander entweder positiv oder negativ auf den extrazellulären, intrazellulären und Transkriptionsebenen 3, 4, 5, 6. Wichtig ist , dass die Kernkomponenten von Signaltransduktionswegen hoch in allen metazoan Spezies konserviert, und bemerkenswert, führen die meisten der wichtigsten Signalwege oft ähnliche Entwicklungsfunktionen in vielen Arten , wenn Organismen von eng verwandten Stämmen insbesondere 7, 8, 9 zu vergleichen, 10, 11. Die Studie der während der Entwicklung Signalgebung ist eine schwierige Aufgabe in jedem Organismus, undmehrere bedeutende Herausforderungen zu studieren Signalwege in den meisten Deuterostomia Modelle (Vertebraten, Invertebraten Chordaten , Kiemenlochtiere und Echinoderme): 1) In Vertebraten gibt es eine große Anzahl von möglichen Liganden und Rezeptor / Co-Modulator - Wechselwirkungen, die intrazelluläre Transduktion Moleküle sowie mögliche Wechselwirkungen zwischen verschiedenen Signalwegen aufgrund der Komplexität des Genoms 12, 13, 14; 2) Die komplexe Morphologie und morphogenetische Bewegungen bei Wirbeltieren oft erschweren funktionelle Interaktionen in und unter Signaltransduktionswege zu interpretieren; 3) Die Analysen in den meisten Nicht-echinoderm invertebrate Deuterostomia Modellarten werden durch kurze Fenster von gravidity mit Ausnahme einiger tunicate Arten 15, 16 begrenzt. DasSeeigel embryo hat einige der oben erwähnten Beschränkungen und bietet viele einzigartige Eigenschaften für eine detaillierte Analyse von Signaltransduktionswegen in vivo durchgeführt wird . Dazu gehören die folgenden: 1) Die relative Einfachheit des Genoms Seeigel reduziert signifikant die Anzahl der möglichen Ligand, Rezeptor / Co-Rezeptor und intrazellulären Transduktion Molekül - Wechselwirkungen 17; 2) Die KNS die Spezifikation und die Strukturierung der Keimblätter und die wichtigsten embryonalen Achsen steuern gut in Seeigel - Embryonen etabliert, in dem Verständnis der regulatorischen Kontext der Zelle / Gebiet Beihilfe Empfangen der Signale 18, 19; 3) Viele Signaltransduktionswege können zwischen frühen Spaltung und Gastrula Stadien untersucht werden , wenn Embryonen von einem einschichtigen Epithel , deren Morphologie bestehen ist einfacher zu analysieren; 4) Die Moleküle beinhaltend in Bahnen in den Seeigeln Signalisierung leicht manipuliert werden; 5) Viele Seeigel sind für 10 bis 11 Monate im Jahr (zB Strongylocentrotus purpuratus und Lytechinus variegatus trächtig). Hier präsentieren wir eine Methode, um systematisch und effizient Komponenten der Signalwege zu charakterisieren, die angeben, und das Muster Gebiete in Seeigel-Embryonen, die Vorteile zu verdeutlichen, dass mehrere wirbellosen Modellsysteme in der Studie von komplexen molekularen Mechanismen bieten.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Translational-blocking morpholino and/or splice-blocking morpholino</td>
			<td>Gene Tools LLC</td>
			<td>Customized</td>
			<td>More information at www.gene-tools.com</td>
		</tr>
		<tr>
			<td>Glycerol</td>
			<td>Invitrogen</td>
			<td>15514-011</td>
			<td></td>
		</tr>
		<tr>
			<td>FITC (dextran fluorescein isothiocyanate)</td>
			<td>Invitrogen, Life Technologies</td>
			<td>D1821</td>
			<td>Make 25mg/mL stock solution</td>
		</tr>
		<tr>
			<td>Paraformaldehyde 16% solution EM Grade</td>
			<td>Electron Microscopy Sciences</td>
			<td>15710</td>
			<td></td>
		</tr>
		<tr>
			<td>MOPS</td>
			<td>Sigma Aldrich</td>
			<td>M1254-250G</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween-20</td>
			<td>Sigma Aldrich</td>
			<td>23336-0010</td>
			<td></td>
		</tr>
		<tr>
			<td>Formamide</td>
			<td>Sigma Aldrich</td>
			<td>47671-1L-F</td>
			<td></td>
		</tr>
		<tr>
			<td>Yeast tRNA</td>
			<td>Invitrogen</td>
			<td>15401-029</td>
			<td></td>
		</tr>
		<tr>
			<td>Normal Goat Serum</td>
			<td>Sigma Aldrich</td>
			<td>G9023-10mL</td>
			<td></td>
		</tr>
		<tr>
			<td>Alkaline Phosphatase-conjugated anti-digoxigenin antibody</td>
			<td>Roche</td>
			<td>11 093 274 910</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetramisole hydrochloride (levamisole)</td>
			<td>Sigma Aldrich</td>
			<td>L9756-5G</td>
			<td></td>
		</tr>
		<tr>
			<td>Tris Base UltraPure</td>
			<td>Research Products Internationall Corp</td>
			<td>56-40-6</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Chloride</td>
			<td>Fisher Scientific</td>
			<td>BP358-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium chloride</td>
			<td>Sigma Aldrich</td>
			<td>7786-30-3</td>
			<td></td>
		</tr>
		<tr>
			<td>BCIP (5-Bromo-4-Chloro-3-indolyl-phosphate</td>
			<td>Roche</td>
			<td>11 383 221 001</td>
			<td></td>
		</tr>
		<tr>
			<td>4 Nitro blue tetrazolium chloride (NBT)</td>
			<td>Roche</td>
			<td>11 383 213 001</td>
			<td></td>
		</tr>
		<tr>
			<td>Dimethyl Formamide</td>
			<td>Sigma Aldrich</td>
			<td>D4551-500mL</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium Chloride</td>
			<td>Sigma Aldrich</td>
			<td>P9541-5KG</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Bicarbonate</td>
			<td>Sigma Aldrich</td>
			<td>S5761-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium Sulfate</td>
			<td>Sigma Aldrich</td>
			<td>M7506-2KG</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcium Chloride</td>
			<td>Sigma Aldrich</td>
			<td>C1016-500G</td>
			<td></td>
		</tr>
	</tbody>
</table>

the power of simplicity: sea urchin embryos as in vivo developmental models for studying complex cell-to-cell signaling network interactions

Remarkably few cell-to-cell signal transduction pathways are necessary during embryonic development to generate the large variety of cell types and tissues in the adult body form. Yet, each year more components of individual signaling pathways are discovered, and studies indicate that depending on the context there is significant cross-talk among most of these pathways. This complexity makes studying cell-to-cell signaling in any in vivo developmental model system a difficult task. In addition, efficient functional analyses are required to characterize molecules associated with signaling pathways identified from the large data sets generated by next generation differential screens. Here, we illustrate a straightforward method to efficiently identify components of signal transduction pathways governing cell fate and axis specification in sea urchin embryos. The genomic and morphological simplicity of embryos similar to those of the sea urchin make them powerful in vivo developmental models for understanding complex signaling interactions. The methodology described here can be used as a template for identifying novel signal transduction molecules in individual pathways as well as the interactions among the molecules in the various pathways in many other organisms.

Im Seeigel Embryo wir haben gezeigt , dass drei verschiedene Wnt - Signalzweige (Wnt / β-Catenin, Wnt / JNK und Wnt / PKC) 4, 25 interact ein Wnt - Signalnetzwerk zu bilden , die von vorne nach hinten (AP) Strukturierung regelt. Eine der wichtigsten Folgen dieser Signalisierungsereignisse besteht darin , dass die anfängliche breit ausgedrückt anterioren Neuroektoderm (ANE) GRN auf ein kleines Gebiet um den vorderen Pol durch d...

Watch this Scientific Journal Video about The Power of Simplicity: Sea Urchin Embryos as in Vivo Developmental Models for Studying Complex Cell-to-cell Signaling Network Interactions at JoVE.com

The Power of Simplicity: Sea Urchin Embryos as in Vivo Developmental Models for Studying Complex Cell-to-cell Signaling Network Interactions

Dieses Video Artikel beschreibt eine einfache In - vivo - Methodik , die verwendet werden können , um systematisch und effizient Komponenten komplexer Signalwege und regulatorische Netzwerke in vielen wirbellosen Embryonen zu charakterisieren.

Die Kraft der Einfachheit: Seeigel Embry als<em&gt; In Vivo</em&gt; Entwicklungsmodelle für komplexe Zell-zu-Zell-Signalnetzwerk Wechselwirkungen Studieren

Remarkably few cell-to-cell signal transduction pathways are necessary during embryonic development to generate the large variety of cell types and ...

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Cell Biology

Developmental Biology

The Power of Simplicity: Sea Urchin Embryos as in Vivo Developmental Models for Studying Complex Cell-to-cell Signaling Network Interactions

Unit 1

Principles of Cell Signaling

SCIENTISTS IN ACTION

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Protokolle für die Beschaffung Zygotic und somatischen Embryonen zur Untersuchung der Regulation der frühen Embryo-Entwicklung in der Modell-Hülsenfrucht<em&gt; Medicago truncatula</em

Early embryogenesis starting from a single cell zygote goes through rapid cell division and morphogenesis, and is morphologically characterized by ...

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Visualisierung der Knoten und Notochordal Platte In Gastrulating Maus-Embryonen mittels Rasterelektronenmikroskopie und ganze Mount Immunfluoreszenz

The post-implantation mouse embryo undergoes major shape changes after the initiation of gastrulation and morphogenesis. A hallmark of morphogenesis is ...

Assessment of Oxidative Damage in the Primary Mouse Ocular Surface Cells/Stem Cells in Response to Ultraviolet-C (UV-C) Damage

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The ocular surface is subjected to regular wear and tear due to various environmental factors. Exposure to UV-C radiation constitutes an occupational ...

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