Name: studying wnt signaling during patterning of conducting airways
Uploaded: 2016-10-16T13:00:00
Description: Watch this Scientific Journal Video about Studying Wnt Signaling During Patterning of Conducting Airways at JoVE.com

Biz histolojik prosedürleri ile Mike Muntifering ve konfokal görüntüleme ile Matt Kofron ve Gail Macke yardım kabul. Bu çalışma kısmen Sağlık-NHLBI National Institutes (DS K01HL115447) tarafından desteklenmiştir.

Hayvanlar patojenden bağımsız koşullarda barındırıldı. Fareler CCHMC Kurumsal Hayvan Bakımı ve Kullanımı Komitesi (Cincinnati, OH ABD) tarafından onaylanan protokollere göre ele alınmıştır. Bu çalışmaları boyunca kullanılan fareler karışık arka muhafaza edilmiştir. 1. Tüm Dağı X-galaktosidaz Boyama <ol><li> CO 2 inhalasyon yoluyla, E14.5 için E11.5 hamile kadın euthanize. CO 2 odasında hayvan yerleştirin CO 2 ile odasını ...

<ol>
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M., Sun, X. <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+promotes+respiratory+progenitor+identity+in+mouse+foregut.">beta-Catenin promotes respiratory progenitor identity in mouse foregut.</a> Proc Natl Acad Sci U S A. 106, 16287-16292 (2009).</li><li>Cornett, B., 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=Wntless+is+required+for+peripheral+lung+differentiation+and+pulmonary+vascular+development.">Wntless is required for peripheral lung differentiation and pulmonary vascular development.</a> Dev Biol. 379, 38-52 (2013).</li><li>Snowball, J., Ambalavanan, M., Whitsett, J., Sinner, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=34;Endodermal+Wnt+signaling+is+required+for+tracheal+cartilage+formation&amp;#34;.">34;Endodermal Wnt signaling is required for tracheal cartilage formation&amp;#34;.</a> Dev Biol. , (2015).</li><li>Shannon, J. 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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesenchyme+specifies+epithelial+differentiation+in+reciprocal+recombinants+of+embryonic+lung+and+trachea.">Mesenchyme specifies epithelial differentiation in reciprocal recombinants of embryonic lung and trachea.</a> Dev Dyn. 212, 482-494 (1998).</li><li>Li, 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=Wnt5a+regulates+Shh+and+Fgf10+signaling+during+lung+development.">Wnt5a regulates Shh and Fgf10 signaling during lung development.</a> Dev Biol. 287, 86-97 (2005).</li><li>Loscertales, M., Mikels, A. J., Hu, J. K., Donahoe, P. K., Roberts, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chick+pulmonary+Wnt5a+directs+airway+and+vascular+tubulogenesis.">Chick pulmonary Wnt5a directs airway and vascular tubulogenesis.</a> Development. 135, 1365-1376 (2008).</li><li>Yin, Y., 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=An+FGF-WNT+gene+regulatory+network+controls+lung+mesenchyme+development.">An FGF-WNT gene regulatory network controls lung mesenchyme development.</a> Dev Biol. 319, 426-436 (2008).</li><li>Shu, W., 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=Wnt/beta-catenin+signaling+acts+upstream+of+N-myc,+BMP4,+and+FGF+signaling+to+regulate+proximal-distal+patterning+in+the+lung.">Wnt/beta-catenin signaling acts upstream of N-myc, BMP4, and FGF signaling to regulate proximal-distal patterning in the lung.</a> Dev Biol. 283, 226-239 (2005).</li><li>Bretholz, A., Morrisey, R., Hoffman, 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=The+use+of+OpdA+in+rat+models+of+organic+phosphorus+(OP)+poisoning.">The use of OpdA in rat models of organic phosphorus (OP) poisoning.</a> Toxicology. 257, (2009).</li><li>Goss, A. M., 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=Wnt2+signaling+is+necessary+and+sufficient+to+activate+the+airway+smooth+muscle+program+in+the+lung+by+regulating+myocardin/Mrtf-B+and+Fgf10+expression.">Wnt2 signaling is necessary and sufficient to activate the airway smooth muscle program in the lung by regulating myocardin/Mrtf-B and Fgf10 expression.</a> Dev Biol. 356, 541-552 (2011).</li><li>Mucenski, M. L., 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=beta-Catenin+is+required+for+specification+of+proximal/distal+cell+fate+during+lung+morphogenesis.">beta-Catenin is required for specification of proximal/distal cell fate during lung morphogenesis.</a> J Biol Chem. 278, 40231-40238 (2003).</li><li>Hyatt, B. A., Shangguan, X., Shannon, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FGF-10+induces+SP-C+and+Bmp4+and+regulates+proximal-distal+patterning+in+embryonic+tracheal+epithelium.">FGF-10 induces SP-C and Bmp4 and regulates proximal-distal patterning in embryonic tracheal epithelium.</a> Am J Physiol Lung Cell Mol Physiol. 287, L1116-L1126 (2004).</li><li>Del Moral, P. M., 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=VEGF-A+signaling+through+Flk-1+is+a+critical+facilitator+of+early+embryonic+lung+epithelial+to+endothelial+crosstalk+and+branching+morphogenesis.">VEGF-A signaling through Flk-1 is a critical facilitator of early embryonic lung epithelial to endothelial crosstalk and branching morphogenesis.</a> Dev Biol. 290, 177-188 (2006).</li><li>Ott, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Confocal+microscopy+in+large+insect+brains:+zinc-formaldehyde+fixation+improves+synapsin+immunostaining+and+preservation+of+morphology+in+whole-mounts.">Confocal microscopy in large insect brains: zinc-formaldehyde fixation improves synapsin immunostaining and preservation of morphology in whole-mounts.</a> J Neurosci Methods. 172, 220-230 (2008).</li><li>Jahrling, N., Becker, K., Dodt, H. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3D-reconstruction+of+blood+vessels+by+ultramicroscopy.">3D-reconstruction of blood vessels by ultramicroscopy.</a> Organogenesis. 5, 145-148 (2009).</li><li>Park, J., 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=Regulation+of+Sox9+by+Sonic+Hedgehog+(Shh)+is+essential+for+patterning+and+formation+of+tracheal+cartilage.">Regulation of Sox9 by Sonic Hedgehog (Shh) is essential for patterning and formation of tracheal cartilage.</a> Dev Dyn. 239, 514-526 (2010).</li><li>Elluru, R. G., Thompson, F., Reece, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fibroblast+growth+factor+18+gives+growth+and+directional+cues+to+airway+cartilage.">Fibroblast growth factor 18 gives growth and directional cues to airway cartilage.</a> Laryngoscope. 119, 1153-1165 (2009).</li><li>Ahnfelt-Ronne, J., 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=An+improved+method+for+three-dimensional+reconstruction+of+protein+expression+patterns+in+intact+mouse+and+chicken+embryos+and+organs.">An improved method for three-dimensional reconstruction of protein expression patterns in intact mouse and chicken embryos and organs.</a> J Histochem Cytochem. 55, 925-930 (2007).</li><li>Yang, B., 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=Single-cell+phenotyping+within+transparent+intact+tissue+through+whole-body+clearing.">Single-cell phenotyping within transparent intact tissue through whole-body clearing.</a> Cell. 158, 945-958 (2014).</li><li>Gillotte, D. M., Fox, P. L., Mjaatvedt, C. H., Hoffman, S., Capehart, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+vitro+method+for+analysis+of+chondrogenesis+in+limb+mesenchyme+from+individual+transgenic+(hdf)+embryos.">An in vitro method for analysis of chondrogenesis in limb mesenchyme from individual transgenic (hdf) embryos.</a> Methods Cell Sci. 25, 97-104 (2003).</li><li>Cohen, E. D., 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=Wnt+signaling+regulates+smooth+muscle+precursor+development+in+the+mouse+lung+via+a+tenascin+C/PDGFR+pathway.">Wnt signaling regulates smooth muscle precursor development in the mouse lung via a tenascin C/PDGFR pathway.</a> J Clin Invest. 119, 2538-2549 (2009).</li><li>Boucherat, O., 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=Partial+functional+redundancy+between+Hoxa5+and+Hoxb5+paralog+genes+during+lung+morphogenesis.">Partial functional redundancy between Hoxa5 and Hoxb5 paralog genes during lung morphogenesis.</a> Am J Physiol Lung Cell Mol Physiol. 304, L817-L830 (2013).</li></ol>

Solunum yollarının morfojenezi yatan olaylar tamamen iletken solunum yollarının desen için gerekli özel işlemler anlaşılamamıştır. Daha önceki çalışmalar, gelişmekte olan eksplantlar hava-sıvı arayüz kültüre ya Matrigel 21,22 gömülü burada eks vivo teknikleri kullanmışlardır. büyüme faktörleri, gelişmekte trakea desenlendirme ve trakeal kıkırdak oluşumunu nasıl etkilediğini Bu çalışmalar göstermiştir. Bu çalışmalara bir sınırlama doku mimarisi düzgün m...

Solunum yolu gelişimi ventral endodermal ön bağırsak 1,2 Nkx2.1 pozitif hücrelerin ortaya çıkması ile embriyonik gün 9 (E9) ile başlatılır. Tüpler ayrı varlıklar olarak ayırt edilebilir zaman Özofagus-trakeal tüp ayırma, E11.5 ile mezenkimal dokudan 3 çevrili her çözecektir. Wnt sinyal akciğer agenezisi 4,5 neden olur endodermal solunum epitel splanknik mesenkim ve β-katenin silinmesi ile ifade Wnt2 ve Wnt2b silinmesi gibi solunum yollarının tarifnamede önemli bir rol oynar. Daha önceki çalışmalar, akciğer hipoplazisi içinde Endodermal solunum yolu sonuçlarından, WLS, tüm Wnt ligandları bir kargo reseptör aracılık salgılanmasının o silinmesini tespit, pulmoner vasküler gelişim ve trakeal mezenşimin 6,7 mis-desen kusurları. Bu veriler epitelyal-mezenkimal cro önemini destekleyenAyrıca, başka çalışmalarda 8,9 gösterildiği gibi p, hücre farklılaşması ve tarifnamede söz. Akciğer gelişiminin erken aşamalarında çalışma bize daha iyi solunum kimliğini 10-16 sürüş mekanizmaları anlamak için izin vitro ve ex vivo teknikleri, genetik dayanmaktadır. Hava sıvı interfaz tüm akciğer eksplant kültürleri yaygın morfolojilerinden 10,17,18 dallanma pulmoner erken dönemlerinde büyüme faktörlerinin etkilerini incelemek için kullanılmıştır. Bu yöntem morfolojik gibi dallanma morfolojilerinden olarak değişiklikler ve gen ifadesi modülasyon okuma olarak kullanılan iken, kültür kendisi damarsal 17 gelişimini desteklemek değil gibi, gelişim sürecinin erken aşamalarında çalışma ile sınırlıdır. trakeal kıkırdak gelişimi bu kültür tekniği ile uyumlu olabilir uzun inkübasyon süreleri gerektirir. onu çözümlemesolunum yolu oluşumu sırasında Wnt sinyal e rolü, bizim embriyonik çalışmaların ihtiyaçlarını karşılamak için standart teknikleri adapte olması. Biz hacimleri, boyama süreleri, trakea-akciğer dokusunun temizlenmesi için parafine ve zamanlama için işlem bisiklet değiştirdiniz. Bu çalışmada açıklanan teknikler optimize ana hedefi E14.5 için E11 gerçekleşecek farelerde trakeal gelişiminin erken aşamalarında analiz etmektir. Muhabir fareler hat Axin2LacZ gelişen trakeal mezenşimin Wnt / β-katenin aktivitesinin biz kesin kararlı siteleri kullanma. Biz de tüm montaj trakeal doku için lektin boyama işlemini adapte var. Böylece, mezenkimal terlemeyi görselleştirmek ve kondrogenezis yer alacak siteleri tahmin başardık. Bütün montaj ve gelişmiş mikroskopi teknikleri ile birleştiğinde WlsShhCre farelerden elde edilen embriyonik doku bölümlerinin, boyanması tra tarafından üretilen Wnt ligandların rolünü açıklayacak için bize izintrakea dokusunda CHEAL epitel.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Anti Sox9 ab.</td>
			<td>Millipore</td>
			<td>AB5535</td>
			<td>1:400 , rabbit</td>
		</tr>
		<tr>
			<td>Anti Sox9 ab.</td>
			<td>Santa Cruz</td>
			<td>Sc-20095</td>
			<td>1:50, rabbit</td>
		</tr>
		<tr>
			<td>Anti Smooth Muscle Actin ab.</td>
			<td>Sigma</td>
			<td>A5228</td>
			<td>1:2k, mouse</td>
		</tr>
		<tr>
			<td>Anti NKX2.1 ab.</td>
			<td>Seven Hills</td>
			<td>n/a</td>
			<td>1:100, guinea pig</td>
		</tr>
		<tr>
			<td>Anti NKX2.1 ab.</td>
			<td>Seven Hills</td>
			<td>n/a</td>
			<td>1:400, mouse</td>
		</tr>
		<tr>
			<td>Anti Brdu ab.</td>
			<td>Abcam</td>
			<td>AB1893</td>
			<td>1:200, sheep</td>
		</tr>
		<tr>
			<td>Anti Brdu ab.</td>
			<td>Santa Cruz</td>
			<td>Sc-32323</td>
			<td>1:4k, mouse</td>
		</tr>
		<tr>
			<td>PNA Lectin</td>
			<td>Sigma</td>
			<td>L 7381</td>
			<td></td>
		</tr>
		<tr>
			<td>Secondary antibodies</td>
			<td>Life technologies</td>
			<td></td>
			<td>Alexa fluor Molecular probes</td>
		</tr>
		<tr>
			<td>K3Fe(CN)6</td>
			<td>Sigma</td>
			<td>P8131</td>
			<td></td>
		</tr>
		<tr>
			<td>K4Fe(CN)6</td>
			<td>Sigma-Aldrich</td>
			<td>P3289</td>
			<td></td>
		</tr>
		<tr>
			<td>MgCl2</td>
			<td>Sigma-Aldrich</td>
			<td>M9272</td>
			<td></td>
		</tr>
		<tr>
			<td>NaDOC</td>
			<td>Life Technologies</td>
			<td>89905</td>
			<td></td>
		</tr>
		<tr>
			<td>NP4O</td>
			<td>Life Technologies</td>
			<td>85124</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcian Blue 8GX</td>
			<td>Sigma</td>
			<td>A-3157</td>
			<td></td>
		</tr>
		<tr>
			<td>Fisher brand super-frost plus</td>
			<td>Fisher</td>
			<td>12-550-15</td>
			<td></td>
		</tr>
		<tr>
			<td>PFA (16%)</td>
			<td>EMS</td>
			<td>15710</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Gibco</td>
			<td>70011-044</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal Calf Serum</td>
			<td>Sigma</td>
			<td>11K413</td>
			<td></td>
		</tr>
		<tr>
			<td>Blocking reagent</td>
			<td>Invitrogen</td>
			<td></td>
			<td>Component of TSA kit #2&#160;&#160;&#160; ( T20932)</td>
		</tr>
		<tr>
			<td>BrDu</td>
			<td>Sigma</td>
			<td>B5002-5g</td>
			<td></td>
		</tr>
		<tr>
			<td>Vectashield mounting medium</td>
			<td>Vector labs</td>
			<td>H-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Permount</td>
			<td>Fisher</td>
			<td>SP15-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue-loc cassettes Histoscreen</td>
			<td>Fisher</td>
			<td>C-0250-GR</td>
			<td></td>
		</tr>
		<tr>
			<td>Biopsy cassettes</td>
			<td>Premiere</td>
			<td>BC0109</td>
			<td>Available in different colors</td>
		</tr>
		<tr>
			<td>Nuclear fast red&#160; Kernechtrot 0.1%</td>
			<td>Sigma</td>
			<td>N3020</td>
			<td></td>
		</tr>
		<tr>
			<td>Citric acid</td>
			<td>Sigma</td>
			<td>C1909-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium citrate tribasic dihydrate</td>
			<td>Sigma</td>
			<td>S4641-1Kg</td>
			<td></td>
		</tr>
		<tr>
			<td>Trizma hydrochloride</td>
			<td>Sigma</td>
			<td>T5941-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylene</td>
			<td>Pharmco-AAPER</td>
			<td>399000000</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Pharmco-AAPER</td>
			<td>111000200</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro knives</td>
			<td>FST</td>
			<td>10318-14</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont #5 ceramic coated</td>
			<td>FST</td>
			<td>11252-50</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont #5CO</td>
			<td>FST</td>
			<td>11295-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont # 5</td>
			<td>FST</td>
			<td>91150-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermo/Shandon Excelsior ES</td>
			<td>Thermo Fisher</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Microtome</td>
			<td>Leica</td>
			<td>RM2135</td>
			<td></td>
		</tr>
		<tr>
			<td>Nikon i90</td>
			<td>Nikon</td>
			<td></td>
			<td>Wide field microscope</td>
		</tr>
		<tr>
			<td>NikonA1Rsi</td>
			<td>Nikon</td>
			<td></td>
			<td>Confocal microscopy. Settings:NikonA1 plus camera, scanner: Galvano, detector:DU4. Optics Plan Apo lambda 10x. Modality: Widefield fluorescence laser confocal.&#160;</td>
		</tr>
		<tr>
			<td>Leica MS 16 FA</td>
			<td>Leica</td>
			<td></td>
			<td>Fluorescence Dissecting microscope</td>
		</tr>
		<tr>
			<td>Zeiss</td>
			<td>Zeiss</td>
			<td></td>
			<td>Automated fluorescence microscope</td>
		</tr>
		<tr>
			<td>Leica Application suite</td>
			<td>Leica</td>
			<td></td>
			<td>Leica imaging software</td>
		</tr>
		<tr>
			<td>NIS</td>
			<td>Nikon</td>
			<td></td>
			<td>Nikon imaging software</td>
		</tr>
		<tr>
			<td>IMARIS</td>
			<td>Bitplane</td>
			<td></td>
			<td>Imaging processing software</td>
		</tr>
	</tbody>
</table>

studying wnt signaling during patterning of conducting airways

Wnt signaling pathways play critical roles during development of the respiratory tract. Defining precise mechanisms of differentiation and morphogenesis controlled by Wnt signaling is required to understand how tissues are patterned during normal development. This knowledge is also critical to determine the etiology of birth defects such as lung hypoplasia and tracheobronchomalacia. Analysis of earliest stages of development of respiratory tract imposes challenges, as the limited amount of tissue prevents the performance of standard protocols better suited for postnatal studies. In this paper, we discuss methodologies to study cell differentiation and proliferation in the respiratory tract. We describe techniques such as whole mount staining, processing of the tissue for confocal microscopy and immunofluorescence in paraffin sections applied to developing tracheal lung. We also discuss methodologies for the study of tracheal mesenchyme differentiation, in particular cartilage formation. Approaches and techniques discussed in the current paper circumvent the limitation of material while working with embryonic tissue, allowing for a better understanding of the patterning process of developing conducting airways.

Wnt / β-katenin etkinliği Tüm montaj Lac-Z boyama raportör Axin2 Lac-Z fareler 11 izole edilmiş embriyolar, trakeal akciğer dokusunda tespit edilmiştir. boyama Yer Wnt / β-katenin etkinliğini gösterir. bütün montaj boyama bölgelerinin analizi Wnt / β-katenin aktivitesi trakea mezenşimin ve gelişmekte olan akciğerlerin periferal bölgelerinde mezenşimin mevcut olduğun...

Watch this Scientific Journal Video about Studying Wnt Signaling During Patterning of Conducting Airways at JoVE.com

Studying Wnt Signaling During Patterning of Conducting Airways

Bütün montaj bağlanmış raportör fareler ve bölüm boyama, mikroskopi ve in vivo deneylerde kullanımı solunum yolu normal model oluşturmanın altında yatan mekanizmaların analizi kolaylaştırır. İşte bu teknikler trakeal gelişimi sırasında Wnt sinyal analizine katkıda açıklar.

İletken Airways Desenlendirme sırasında Wnt Sinyalizasyonu incelenmesi

Wnt signaling pathways play critical roles during development of the respiratory tract. Defining precise mechanisms of differentiation and morphogenesis ...

Research

JoVE Journal

Developmental Biology

Method of Studying Palatal Fusion using Static Organ Culture

Statik Organ Kültürü kullanarak Palatal Fusion incelenmesi yöntemi

Cleft lip and palate are among the most common of all birth defects. The secondary palate forms from mesenchymal shelves covered with epithelium that ...

Using Confocal Analysis of Xenopus laevis to Investigate Modulators of Wnt and Shh Morphogen Gradients

Using Confocal Analysis of Xenopus laevis to Investigate Modulators of Wnt and Shh Morphogen Gradients

Konfokal Analizi Kullanımı<em&gt; Xenopus laevis</em&gt; Wnt ve Shh morfogen Gradyanların modülyatorlar Soruşturma

This protocol describes a method to visualise ligands distributed across a field of cells. The ease of expressing exogenous proteins, together with the ...

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

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

Sadelik Gücü: olarak deniz kestanesi Embriyolar<em&gt; Vivo</em&gt; Karmaşık Hücre-hücre Sinyal Ağ Etkileşimleri incelenmesi için Gelişim Modelleri

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

Stimulation of Notch Signaling in Mouse Osteoclast Precursors

Fare Osteoklast Öncüleri Notch Sinyal uyarılması

Notch signaling is a key component of multiple physiological and pathological processes. The nature of Notch signaling, however, makes in vitro ...

In Vitro Ovule Cultivation for Live-cell Imaging of Zygote Polarization and Embryo Patterning in Arabidopsis thaliana

In Vitro Ovule Cultivation for Live-cell Imaging of Zygote Polarization and Embryo Patterning in Arabidopsis thaliana

Vitro Zigot polarizasyon, Live-hücre görüntüleme ve Arabidopsis thaliana embriyo desenlendirme için ovule ekimi

In most flowering plants, the zygote and embryo are hidden deep in the mother tissue, and thus it has long been a mystery of how they develop dynamically; ...

Live Cell Imaging of Chromosome Segregation During Mitosis

Kromozom segregasyon mitoz sırasında canlı hücre görüntüleme

Chromosomes must be reliably and uniformly segregated into daughter cells during mitotic cell division. Fidelity of chromosomal segregation is controlled ...

Visualization of the Superior Ocular Sulcus during Danio rerio Embryogenesis

Visualization of the Superior Ocular Sulcus during Danio rerio Embryogenesis

Çift oküler sulkus görselleştirme Danio rerio embriyogenez sırasında

Congenital ocular coloboma is a genetic disorder that is typically observed as a cleft in the inferior aspect of the eye resulting from incomplete choroid ...

Genotyping of Sea Anemone during Early Development

Erken gelişim sırasında deniz anemon Genotipleme

Described here is a PCR-based protocol to genotype the gastrula stage embryo of the anthozoan cnidarian Nematostella vectensis without sacrificing the ...

A Layered Mounting Method for Extended Time-Lapse Confocal Microscopy of Whole Zebrafish Embryos

Tüm Zebrabalığı Embriyolarının Uzun Zaman Atlamalı Konfokal Mikroskobu için Katmanlı Montaj Yöntemi

Dynamics of development can be followed by confocal time-lapse microscopy of live transgenic zebrafish embryos expressing fluorescence in specific tissues ...

Real-time Bioluminescence Imaging of Notch Signaling Dynamics during Murine Neurogenesis

Murin Nörogenezi Sırasında Çentik Sinyal Dinamiğinin Gerçek Zamanlı Biyolüminesans Görüntülemesi

Notch signaling regulates the maintenance of neural stem/progenitor cells by cell-cell interactions. The components of Notch signaling exhibit dynamic ...