Name: real-time imaging of heterotypic platelet-neutrophil interactions on the activated endothelium during vascular inflammation and thrombus formation in live mice
Uploaded: 2013-04-02T13:00:00
Description: Watch this Scientific Journal Video about Real-time Imaging of Heterotypic Platelet-neutrophil Interactions on the Activated Endothelium During Vascular Inflammation and Thrombus Formation in Live Mic

Bu çalışma Ulusal Sağlık Enstitüleri ile hibe (JC P30 HL101302 ve RO1 HL109439) ve Amerikan Kalp Derneği (JC SDG 5270005) tarafından kısmen desteklenmiştir. A. Barazia bir T32HL007829 NIH eğitim hibesi ile desteklenmiştir.

1. Intravital Mikroskop hazırlanması (Şekil 1A) <ol><li> (125 mM NaCI, 4.5 mM KCI, 2.5 mM CaCl2, 1 mM MgCl2, 17 mM NaHCO 3, pH 7.4) tampon superfusion hazırlayın. </li><li> 37 tampon sıcaklık ve bir termo-kontrollü battaniye korumak için bir dolaşım su banyosu açın ° C. Azot gazı (% 5 CO 2, nitrojen ile dengeli) ile tampon havalandırınız. </li><li> Mikroskop sistemi açın (Sutter Lambda DG-4 yüksek hızlı dalga değiştirici, iş istasyonu bilgisayar, Ol...

<ol>
	<li>Wagner, D. D., Frenette, P. 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+vessel+wall+and+its+interactions.">The vessel wall and its interactions.</a> Blood. 111, 5271-5281 (2008).</li><li>Nieswandt, B., Kleinschnitz, C., Stoll, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ischaemic+stroke:+a+thrombo-inflammatory+disease.">Ischaemic stroke: a thrombo-inflammatory disease.</a> J. Physiol. 589, 4115-4123 (2011).</li><li>Yang, J., Furie, B. 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R., Furie, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+critical+role+for+extracellular+protein+disulfide+isomerase+during+thrombus+formation+in+mice.">A critical role for extracellular protein disulfide isomerase during thrombus formation in mice.</a> J. Clin. Invest. 118, 1123-1131 (2008).</li><li>Cho, 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=Protein+disulfide+isomerase+capture+during+thrombus+formation+in+vivo+depends+on+the+presence+of+beta3+integrins.">Protein disulfide isomerase capture during thrombus formation in vivo depends on the presence of beta3 integrins.</a> Blood. 120, 647-655 (2012).</li><li>Gross, P. L., Furie, B. 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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=Alpha+1,3+fucosyltransferases+are+master+regulators+of+prostate+cancer+cell+trafficking.">Alpha 1,3 fucosyltransferases are master regulators of prostate cancer cell trafficking.</a> Proceedings of the National Academy of Sciences of the United States of America. 106, 19491-19496 (2009).</li><li>Trzpis, M., McLaughlin, P. M., de Leij, L. M., Harmsen, M. 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Burada vasküler inflamasyon ve tromboz sırasında aktive endotel üzerindeki heterotipik trombosit-nötrofil etkileşimleri görselleştirmek için gerçek zamanlı floresan intravital mikroskopi için ayrıntılı bir protokol açıklar. Daha önce, benzer floresans mikroskopik yaklaşımlar trombüs oluşumu ve vasküler inflamasyonun moleküler mekanizması çalışmaya bildirildi. 8,12 heterotipik hücre-hücre etkileşimi yaralanma yerinde vazo-oklüzyon için önemli olabilir bu yana, bu teknoloji iç...

<table border="1">
	<tbody>
		<tr>
			<td>Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Fisher Scientific</td>
			<td>7647-14-5</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>KCl</td>
			<td>Sigma-Aldrich</td>
			<td>7447-40-7</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>CaCl2 2H2O</td>
			<td>Sigma-Aldrich</td>
			<td>10035-04-8</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>MgCl2 6H2O</td>
			<td>Fisher Scientific</td>
			<td>7791-18-6</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>NaHCO3</td>
			<td>Fisher Scientific</td>
			<td>144-55-8</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>0.9% NaCl Saline</td>
			<td>Hospira</td>
			<td>0409-4888-10</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Hospira</td>
			<td>0409-2051-05</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Lloyd</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Intramedic Tubing (PE 90)</td>
			<td>BD Diagnostics</td>
			<td>427421</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Intramedic Tubing (PE 10)</td>
			<td>BD Diagnostics</td>
			<td>427401</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Murine TNF-&alpha;</td>
			<td>R&amp;D Systems</td>
			<td>410-MT</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Dylight 488- labeled rat anti-mouse CD42b antibody</td>
			<td>Emfret Analytics</td>
			<td>X488</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Alexa Fluor 647-conjugated anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody</td>
			<td>BioLegend</td>
			<td>108418</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>NESLAB EX water bath/circulator</td>
			<td>Thermo-Scientific</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Olympus BX61W microscope</td>
			<td>Olympus</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>TH4-100 Power</td>
			<td>Olympus</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Lambda DG-4</td>
			<td>Sutter</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>MPC-200 multi-manipulator</td>
			<td>Sutter</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>ROE-200 stage controller</td>
			<td>Sutter</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>C9300 high-speed camera</td>
			<td>Hamamatsu</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Intensifier</td>
			<td>Video Scope International</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Ablation Laser</td>
			<td>Photonic Instruments, Inc.</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>SlideBook 5.0</td>
			<td>Intelligent Imaging Innovations</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
	</tbody>
</table>

real-time imaging of heterotypic platelet-neutrophil interactions on the activated endothelium during vascular inflammation and thrombus formation in live mice

Arterioler yaralanma yerinde trombüs oluşumu sırasında aktive endotel aracılık tromboz ve vasküler inflamasyon üzerine aktive trombositler ve lökositler (özellikle nötrofiller) etkileşimi. 1,2, aktive endotel ve subendotelyal matriks proteinlerine yapışkan trombositlere nötrofil yuvarlanma ve yapışma destekliyoruz. 3 Tersine, venüler inflamatuar koşullarda, aktif endotele yapıştığında nötrofil trombositlerin dolaşımdaki adezyon ve birikimi destekleyebilir. Heterotipik trombosit-agregasyonu nötrofil hücreleri arasındaki spesifik reseptör-reseptör etkileşimleri yoluyla sayaç ardışık işlemler gerektirir. 4 O aktive edilmiş endotel hücreleri, böylece yüksek kesme koşulları altında trombosit yapışması ve birikim başlatılması, bu tür von Willebrand faktörü gibi yapışma molekülleri serbest olduğu bilinmektedir. 5 Ayrıca, aktive endotel hücreleri ifade selektinler nötrofil yuvarlanma ve adezyon bir destekd Hücrelerarası adezyon molekülü-1 (ICAM-1), sırasıyla, düşük kesme koşulları altında. 4 Trombosit P-selektin yoluyla nötrofiller ile etkileşime girer p-selektin glikoprotein ligandı-1 (PSGL-1), nötrofil ve böylece β2 integrinler ve sağlam aktivasyonunu uyararak İki hücre türleri arasındaki yapışma. Heterotipik trombosit-nötrofil etkileşimleri tam kan ya da izole edilmiş hücreler 6,7 olarak belirlendiği in vitro deneylerde gelişmelere rağmen, bu çalışmalar vasküler hastalık sırasında oksidatif stres koşulları manipüle edemez. Bu raporda, trombosit ve nötrofil işaretleyici bir fare karşı floresan-etiketli, spesifik antikorlar kullanılarak, TNF-α ile indüklenen inflamasyonu veya lazer kaynaklı takip sırasında aktive endotel üzerinde trombosit ve nötrofil heterotipik etkileşimlerini izlemek için ayrıntılı bir intravital mikroskopik protokol açıklar canlı farelerin cremaster kas mikrodamar yaralanma.

Ayrıntılı bir intravital mikroskopi analizi kullanarak, aktive endotel üzerindeki heterotipik trombosit-nötrofil etkileşimleri trombosit (CD42c) veya nötrofil işaretleyici (Gr-1) canlı farelere karşı floresan etiketli antikorlar infüzyonu ile görüntülenmiştir. TNF-α-kaynaklı venüler enflamasyon modelinde, en çok haddeleme nötrofiller stabil kayıt süresi (3-4,5 saat TNF-α enjeksiyonundan sonra, Şekil 2A&#39;da sırasında ICAM-1 ile aktive β2 İntegr...

Watch this Scientific Journal Video about Real-time Imaging of Heterotypic Platelet-neutrophil Interactions on the Activated Endothelium During Vascular Inflammation and Thrombus Formation in Live Mic

Real-time Imaging of Heterotypic Platelet-neutrophil Interactions on the Activated Endothelium During Vascular Inflammation and Thrombus Formation in Live Mice

Burada vasküler inflamasyon ve canlı farelerde trombüs oluşumu sırasında aktive endotel üzerindeki heterotipik trombosit-nötrofil etkileşimleri görselleştirmek için floresan intravital mikroskobu deneysel bir tekniktir rapor. Bu mikroskobik teknoloji vasküler hastalığın moleküler mekanizmasını incelemek ve patofizyolojik durumlardaki farmakolojik ajanları test için değerli olacaktır.

Canlı Fare Damar İltihabı ve trombüs oluşumu sırasında Activated Endotel üzerinde heterotypic Trombosit nötrofil Etkileşimleri Gerçek zamanlı Görüntüleme

Interaction of activated platelets and leukocytes (mainly neutrophils) on the activated endothelium mediates thrombosis and vascular inflammation.1,2 ...

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Deciphering and Imaging Pathogenesis and Cording of Mycobacterium abscessus in Zebrafish Embryos

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Zebrafish (Danio rerio) embryos are increasingly used as an infection model to study the function of the vertebrate innate immune system in host-pathogen ...

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In Vitro Mikroakışkan Hastalık Modeli Gerçek Zamanlı Olarak Tam Kan-Endotel Etkileşimleri ve Kan Pıhtısı Dinamikleri Çalışma

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