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. Høy gjennomstrømning Morpholino Design Strategy <ol><li> Identifisere et gen (er) av interesse (f.eks kandidat genet tilnærming, cis-regulatoriske analyse, RNAseq og / eller proteomikk differensial skjermer). </li><li> Bruk genomisk, transcriptomic, og genekspresjon data tilgjengelig på hyppig oppdaterte nettsider (f.eks SpBase http://www.echinobase.org 20 og S. purpuratus Genome Søk http: ///urchin.nidcr.nih.gov/blast/index html) for å fastslå at spatiotemp...

<ol>
	<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>

Metodikken som presenteres her er et eksempel som illustrerer kraften i å bruke embryo med mindre genomisk og morfologisk kompleksitet enn virveldyr å forstå signaloverføringsveier og GRNs regulerer grunnleggende utviklingsmekanismer .. Mange laboratorier bruker lignende undersøkelser i løpet av tidlig kråkeboller utvikling for å dissekere signalveier involvert i andre arrangementer celle skjebne spesifikasjon (f.eks Notch, pinnsvin, TGF-β, og FGF signalering) 27, <su...

Gennettverk (GRNs) og signaltransduksjonsveiene etablere den romlige og tidsmessige ekspresjon av genene under embryonal utvikling som brukes til å bygge opp den voksne dyrekroppen plan. Celle-til-celle signaltransduksjonsveier er viktige komponenter i disse regulatoriske nettverk, noe som gir et middel som celler kommuniserer. Disse cellulære interaksjoner etablere og videreutvikle et uttrykk for regulatoriske og differensiering gener i og mellom de ulike områdene under embryogenese 1, 2. Interaksjoner mellom utskilte ekstracellulære modulatorer (ligander, antagonister), reseptorer, og co-reseptorer kontrollere aktiviteter signaltransduksjonsveiene. Et utvalg av intracellulære molekyler transduces disse inngangene resulterer i forandret genekspresjon, divisjon, og / eller formen av en celle. Mens mange av de viktigste molekylene som brukes på ekstracellulære og intracellulære nivåer i de store banene erkjent, er det en ufullstendig kjennskap skyldes i stor grad til kompleksiteten av de enkelte signalveier. I tillegg er forskjellige signalbaner ofte kommuniserer med hverandre enten positivt eller negativt i det ekstracellulære, intracellulære, og transkripsjonelle nivå 3, 4, 5, 6. Viktigere er de viktigste komponentene i signaloverføringsveier høyt konservert i alle metazo arter, og bemerkelsesverdig, de fleste av de store signalveier ofte utføre lignende utviklingsfunksjoner i mange arter når man sammenligner organismer fra nært beslektet phyla spesielt 7, 8, 9, 10, 11. Studiet av signale under utvikling er en krevende oppgave i enhver organisme, og deter flere store utfordringer til å studere signalveier i de fleste deuterostomier modeller (virveldyr, virvelløse chordatar, hemichordater og pigghuder): 1) I virveldyr er det et stort antall mulige ligand og reseptor / co-modulator interaksjoner, intracellulære transduksjon molekyler, samt potensielle interaksjoner mellom forskjellige signalveier på grunn av kompleksiteten av genomet 12, 13, 14; 2) Et kompleks morfologi og morfogenetiske bevegelser hos virveldyr ofte gjøre det vanskeligere å tolke funksjonelle interaksjoner i og mellom signaloverføringsreaksjonsveier; 3) Analyser i de fleste ikke-echinoderm virvelløse deuterostomier modell arter er begrenset av korte vinduer av graviditet med unntak av noen kappe arter 15, 16. Dekråkeboller embryo har noen av de ovenfor nevnte begrensninger, og byr på mange unike egenskaper for å utføre en detaljert analyse av signaltransduksjonsveier in vivo. Disse inkluderer følgende: 1) Den relative enkelhet av kråkeboller genomet reduserer antall mulige ligand, reseptor / co-reseptor og intracellulær transduksjon molekyl betydelig interaksjoner 17; 2) De GRNs kontrollere spesifikasjonen og fordelingen av kjønns lag og store embryonale akser er godt etablert i kråkebolle embryoer, hjelpe i forståelsen av regelverket sammenheng med celle / territorium mottar signalene 18, 19; 3) mange signaltransduksjonsveiene kan studeres mellom tidlige spaltningsseter og gastrula stadier når embryoer bestå av et enkelt lag epitelet hvis morfologi er lettere å analysere; 4) Molekylene involvered i signalveier i kråkebollene er lett manipulert; 5) Mange kråkeboller er gravid i 10 til 11 måneder i året (f.eks Strongylocentrotus purpuratus og Lytechinus variegatus). Her presenterer vi en metode for å systematisk og effektivt karakter komponenter av signalveier som spesifiserer og mønster territorier i kråkebolle embryo for å illustrere fordelene at flere virvelløse modellsystemer tilbyr i studiet av komplekse molekylære mekanismer.

<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.

I kråkebolle embryo har vi vist at 3 forskjellige Wnt signale grener (Wnt / β-catenin, Wnt / JNK, og Wnt / PKC) 4, 25 samhandle for å danne en Wnt signalnettverk som styrer anterior-posterior (AP) mønster. En av de viktigste konsekvensene av disse signal hendelsene er at den første bredt uttrykt anterior neuroectoderm (ANE) grn blir begrenset til et lite område rundt fremre stang ved begynnelsen av gastrulation (24 HPF i <e...

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

Denne videoen artikkelen detaljer en grei in vivo metodikk som kan brukes til systematisk og effektivt karakter komponenter i komplekse signalveier og regulatoriske nettverk i mange virvelløse embryoer.

The Power of Simplicity: Sea Urchin embryo som<em&gt; I Vivo</em&gt; Utviklings modeller for å studere Complex Cell-til-celle signaleringsnett Interactions

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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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

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