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. Yüksek Verimli Morfolino Tasarım Stratejisi <ol><li> Bir ilgi gen (ler) (örneğin aday gen yaklaşımı, cis-düzenleyici analiz, RNAseq ve / veya proteomik diferansiyel ekranları) belirleyin. </li><li> Sık sık güncellenen web sitelerinde mevcut genomik, transkriptomik ve gen ifadesi verileri kullanın (örn SpBase http://www.echinobase.org 20 ve S. purpuratus Genom Arama http: ///urchin.nidcr.nih.gov/blast/index .html) uzaysal ekspresyon profili söz konusu...

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Burada sunulan metodoloji .. Birçok laboratuvar incelemek için erken deniz kestanesi gelişimi sırasında benzer deneyleri kullanan temel gelişimsel mekanizmaları düzenleyen sinyalizasyon iletim yollarını ve GRNs anlamak için omurgalılar daha az genomik ve morfolojik karmaşıklığı ile embriyoları kullanılarak gücünü gösteren bir örnektir diğer hücre kaderi şartname olaylarda yer alan sinyal yolları (örn Notch, Kirpi, TGF β, ve FGF sinyal) 27, <sup cl...

Gen düzenleyici ağları (GRNs) ve sinyal iletim yolları yetişkin hayvan vücut planı oluşturmak için kullanılan embriyonik gelişim sırasında genlerin mekansal ve zamansal ifade kurmak. Hücre-hücre sinyal iletim yolları hücrelerin iletişim hangi araçları sağlayan bu düzenleme ağların temel bileşenleri vardır. Bu hücresel etkileşimler kurmak ve embriyogenez 1, 2 sırasında çeşitli bölgelerde ve arasında düzenleyici ve farklılaşma genlerinin ekspresyonunu rafine. salgılanmış hücre dışı modülatörlerinin (ligandlar, antagonistler), reseptör ve ko-reseptörleri arasındaki etkileşimi sinyal transdüksiyon yollarının aktivitelerini kontrol eder. hücre içi moleküller bir yelpazesine bir hücrenin gen ekspresyonu değişimlerinin, bölme ve / veya şekli ile sonuçlanan, bu giriş çeviren. ana yoldan büyük hücre dışı ve hücre içi seviyede kullanılan anahtar moleküllerinin çoğu, birliktebilinen, tek tek sinyal yollarının karmaşıklığı nedeniyle büyük ölçüde eksik bilgidir. Buna ek olarak, farklı sinyal yolları çoğunlukla dışı, hücre içi de olumlu veya olumsuz ya birbiri ile etkileşim ve transkripsiyon seviyeleri 3, 4, 5, 6. Önemli olarak, sinyal transdüksiyon yollarının ana bileşenleri çok tüm metazoan türlerinde korunan ve özellikle yakından ilgili filumlar organizmaların karşılaştırırken, son derece önemli sinyal yollarının en sık birçok türün içindeki gelişim işlevleri yerine 7, 8, 9, 10, 11. gelişimi sırasında sinyal çalışma herhangi bir organizmada zor bir görev olduğunu ve oradaomurgalılarda büyük olası ligand ve reseptör / ko-modülatör etkileşimlerinin sayılar, hücre içi iletim molekülleri, hem de orada 1): en ikincil ağızlılar modelleri (omurgalıların omurgasız omurgalılar, yarı sırtipliler ve derisidikenlilerde) &#39;de sinyal yolları okuyan bazı önemli zorluklar nedeniyle genom 12, 13, 14 karmaşıklığı farklı sinyaller arasındaki etkileşimler; 2) omurgalıların karmaşık morfolojisi ve morfogenetik hareketler genellikle daha zor ve sinyal iletim yollarının arasında fonksiyonel etkileşimleri yorumlamak yapmak; 3) En dışı derisi dikenliler omurgasız ikincil ağızlılar modeli türlerinde Analizleri bazı tunikat türlerinin 15, 16 hariç gravida kısa pencereler ile sınırlıdır. Deniz kestanesi embriyo yukarıda belirtilen sınırlamaları birkaç vardır ve in vivo sinyal iletim yollarının ayrıntılı bir analiz yapmak için birçok benzersiz niteliklere sahiptir. Bunlar aşağıdakileri içerir: 1), deniz kestanesi genomunun uygulama kolaylığı önemli ölçüde mümkündür ligand, reseptör / co-reseptör ve hücre içi iletim molekülü sayısını azaltır 17 etkileşimi; 2) germ ve büyük embriyonik eksenlerin özellikleri ve desenlendirme kontrol GRNs iyi sinyaller 18, 19 alıcı hücre / topraklarının düzenleyici bağlamında anlamada yardımcı, deniz kestanesi embriyolarda kurulur; Embriyolar olan morfoloji analiz etmek kolay bir tek katmanlı epiteli oluşur zaman 3) Birçok sinyal iletim yolları erken bölünme ve gastrula aşamaları arasında ele alınabilir; 4) molekülleri içerenkolayca manipüle edilir deniz kestanesi sinyal yolları d; 5) Birçok deniz kestanesi 10 ila 11 ay bir yıl (örneğin Strongylocentrotus purpuratus ve Lytechinus variegatus) için gebe edilir. Burada, sistematik ve verimli bir şekilde belirtmek ve deniz kestanesi embriyolarda desen bölgeleri birkaç omurgasız model sistemler karmaşık moleküler mekanizmaların çalışmada sunduğumuz avantajları göstermek için sinyal yollarının bileşenlerini karakterize bir yöntem mevcut.

<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.

Deniz kestanesi embriyo biz 3 farklı Wnt sinyal dalları (Wnt / β-katenin, Wnt / JNK ve Wnt / PKC) 4, 25 etkileşim ön-arka (AP) desenlendirme yöneten bir Wnt sinyal ağı oluşturmak için olduğunu göstermiştir. Bu sinyal olayların en önemli sonuçlarından biri, ilk geniş ifade ön nöroektoderm (ANE) GRN gastrulasyon başında (S. purpuratus 24 hpf) tarafından ön direğin etrafında küçük bir bölgede s...

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

Bu video makalede sistematik ve verimli birçok omurgasız embriyolar karmaşık sinyal yolları ve düzenleyici ağları bileşenleri karakterize etmek için kullanılabilir in vivo metodolojisi basit ayrıntıları.

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 ...

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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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Bone-forming osteoblasts interact with bone-resorbing osteoclasts to coordinate the turnover of bone matrix and to control skeletal homeostasis. Medaka ...

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

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