Name: noninvasive intratracheal intubation to study the pathology and physiology of mouse lung
Uploaded: 2013-11-08T22:00:00
Description: Watch this Scientific Journal Video about Noninvasive Intratracheal Intubation to Study the Pathology and Physiology of Mouse Lung at JoVE.com

Dette arbejde blev støttet af Intramural Research Program for National Cancer Institute, Center for Cancer Research.

Følgende protokol beskriver en ikke-invasiv, enkel og hurtig metode til at levere materialer af interesse for muselunger. Denne procedure blev godkendt af National Cancer Institute Animal Care og brug Udvalg. 1.. Anæstesi <ol><li> Først bedøver musen under anvendelse af en blanding af ketamin og xylazin (100 mg / kg legemsvægt og 10 mg / kg legemsvægt, henholdsvis). Dette er ACUC anbefalede anæstesi og dosis. Med dette beløb, mus er bevidstløs mindste for ~ 20 min. </li><li> Påf?...

<ol>
	<li>Das, S., Macdonald, K., Chang, H. Y., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+of+mouse+lung+intubation.">A simple method of mouse lung intubation.</a> J. Vis. Exp. (73), e50318 (2013).</li><li>Miller, M. 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=Visualization+of+murine+intranasal+dosing+efficiency+using+luminescent+Francisella+tularensis:+effect+of+instillation+volume+and+form+of+anesthesia.">Visualization of murine intranasal dosing efficiency using luminescent Francisella tularensis: effect of instillation volume and form of anesthesia.</a> PLoS One. 7, e31359 (2012).</li><li>Rayamajhi, M., Redente, E. F., Condon, T. V., Gonzalez-Juarrero, M., Riches, D. W., Lenz, L. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-surgical+intratracheal+instillation+of+mice+with+analysis+of+lungs+and+lung+draining+lymph+nodes+by+flow+cytometry.">Non-surgical intratracheal instillation of mice with analysis of lungs and lung draining lymph nodes by flow cytometry.</a> J. Vis. Exp. (51), e2702 (2011).</li><li>Riesenfeld, E. P., 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=Inhaled+salmeterol+and/or+fluticasone+alters+structure/function+in+a+murine+model+of+allergic+airways+disease.">Inhaled salmeterol and/or fluticasone alters structure/function in a murine model of allergic airways disease.</a> Respir. Res. 11, 22 (2010).</li><li>Rivera, B., Miller, S., Brown, E., Price, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+method+for+endotracheal+intubation+of+mice+and+rats+used+in+imaging+studies.">A novel method for endotracheal intubation of mice and rats used in imaging studies.</a> Contemp. Top. Lab Anim. Sci. 44, 52-55 (2005).</li><li>Bonniaud, P., 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=Progressive+transforming+growth+factor+beta1-induced+lung+fibrosis+is+blocked+by+an+orally+active+ALK5+kinase+inhibitor.">Progressive transforming growth factor beta1-induced lung fibrosis is blocked by an orally active ALK5 kinase inhibitor.</a> Am. J. Res. Crit. Care Med. 171, 889-898 (2005).</li><li>Bivas-Benita, M., Zwier, R., Junginger, H. E., Borchard, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+pulmonary+aerosol+delivery+in+mice+by+the+endotracheal+route.">Non-invasive pulmonary aerosol delivery in mice by the endotracheal route.</a> Eur. J. Pharm. Biopharm. 61, 214-218 (2005).</li><li>Haegens, 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=Myeloperoxidase+deficiency+attenuates+lipopolysaccharide-induced+acute+lung+inflammation+and+subsequent+cytokine+and+chemokine+production.">Myeloperoxidase deficiency attenuates lipopolysaccharide-induced acute lung inflammation and subsequent cytokine and chemokine production.</a> J. Immunol. 182, 7990-7996 (2009).</li><li>Jackson, P., 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=Exposure+of+pregnant+mice+to+carbon+black+by+intratracheal+instillation:+toxicogenomic+effects+in+dams+and+offspring.">Exposure of pregnant mice to carbon black by intratracheal instillation: toxicogenomic effects in dams and offspring.</a> Mutat. Res. 745, 73-83 (2012).</li><li>Bodewes, 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=Pathogenesis+of+Influenza+A/H5N1+virus+infection+in+ferrets+differs+between+intranasal+and+intratracheal+routes+of+inoculation.">Pathogenesis of Influenza A/H5N1 virus infection in ferrets differs between intranasal and intratracheal routes of inoculation.</a> Am. J. Pathol. 179, 30-36 (2011).</li><li>Winslow, M. 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=Suppression+of+lung+adenocarcinoma+progression+by+Nkx2-1.">Suppression of lung adenocarcinoma progression by Nkx2-1.</a> Nature. 473, 101-104 (2011).</li></ol>

Intratrakeal intubation beskrevet her er en enkel, men alligevel fremragende noninvasive metode til at jævnt levere materialer af interesse for mus lunge. Denne fremgangsmåde muliggør undersøgelse af effekten og / eller rolle materiale administreret på fysiologi og / eller patologi af lungen. De materialer, der administreres kan være endogene molekyler såsom cytokiner eller eksogene materialer såsom miljøfremmede kemikalier / stoffer, kræftfremkaldende stoffer, forurenende stoffer, allergener, eller virus, der...

Lung er et organ, hvor mange ødelæggende sygdomme er almindeligt diagnosticeret. Blandt dem, lungekræft er den næstmest diagnosticeret kræft hos både mænd og kvinder, og den mest almindelige årsag til kræft død. Kronisk obstruktiv lungesygdom, også kendt som emfysem og kronisk bronkitis, er en meget alvorlig sygdom, og den tredje hyppigste årsag til død i USA. I 2011 blev det anslået, at 25,9 mio amerikanere havde astma, herunder 7,1 millioner børn under 18 år. Astma er den tredje hyppigste årsag til hospitalsindlæggelse blandt børn under 15 år (American Lung Association, <a href="http://www.lung.org" target="_blank">http://www.lung.org</a> ). For at studere patofysiologien og / eller ætiologi af disse ødelæggende sygdomme og deres underliggende mekanismer, anvendelse af nøjagtige modeller er kritisk i forbindelse med praktisk og ikke-invasiv administration af forskellige materialer af interesse for lungen. Mus giver en af ​​de bedste modeller tilstudere de molekylære mekanismer, der ligger patologi og fysiologi lungesygdomme på grund af den øgede tilgængelighed af knockout og transgene mus og et stort mængden af ​​tilgængelig genetisk information. Forskellige metoder er blevet brugt af en række af efterforskere i forskellige indstillinger for at levere materiale af interesse for muselunger, herunder indånding, intranasal instillation, intratrakeal instillation, og intratrakeal intubation 1-4. Sidstnævnte procedure er ikke blevet rutinemæssigt anvendes, fordi det anses temmelig vanskeligt at udføre. Intratracheal intubation beskrevet heri er en ikke-invasiv, enkel og hurtig metode til at levere materialer af interesse for muselunger med henblik på at undersøge virkningen af de leverede materialer på genekspression mønstre, patologi og / eller fysiologi lunge 5. Denne teknik sikrer levering af udefra kommende materiale til en hel lunge og ikke indebærer nogen overlevelse kirurgi, og dermed vil likely skal godkendes af enhver form for institutionelle dyr pleje og brug udvalg.

<table border="1">
	<tbody>
		<tr>
			<td>KetaVed</td>
			<td>Vedco</td>
			<td></td>
			<td>Ketamine</td>
		</tr>
		<tr>
			<td>AnaSed Injection 20 mg</td>
			<td>LLOYD</td>
			<td></td>
			<td>Xylazine</td>
		</tr>
		<tr>
			<td>BioLite Stand</td>
			<td>Braintree Scientific</td>
			<td>RIS100</td>
			<td>For mice</td>
		</tr>
		<tr>
			<td>BioLite Intubation Illumination System</td>
			<td>Braintree Scientific</td>
			<td>BIO MI-KIT</td>
			<td>For mice</td>
		</tr>
		<tr>
			<td>22 G, 1 inch i.v. catheter</td>
			<td>Terumo</td>
			<td>SR-OX2225CA</td>
			<td>For mice below 30 g.&#160;Catheter is used only for the case where the Intubation Illumination System is not used.</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>or</td>
		</tr>
		<tr>
			<td>20 G, 1 inch i.v. catheter</td>
			<td>Terumo</td>
			<td>SR-OX2025CA</td>
			<td>For mice over 30 g.&#160;Catheter is used only for the case where the Intubation Illumination System is not used.</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Catheter is used only for the case where the Intubation Illumination System is not used</td>
		</tr>
	</tbody>
</table>

noninvasive intratracheal intubation to study the pathology and physiology of mouse lung

Anvendelsen af ​​en model, der efterligner tilstanden af ​​lungesygdomme hos mennesker er afgørende for at studere patofysiologi og / eller ætiologi af en bestemt sygdom, og for udvikling af terapeutisk intervention. Med den øgede tilgængelighed af knockout og transgene derivater, sammen med en stor mængde af genetisk information, mus giver en af ​​de bedste modeller til at studere de molekylære mekanismer, der ligger patologi og fysiologi lungesygdomme. Indånding, intranasal instillation, intratrakeal instillation, og intratrakeal intubation er de mest anvendte teknikker ved en række forskere til at administrere materialer af interesse for muselunger. Der er fordele og ulemper for hver teknik, afhængigt af målene for en undersøgelse. Her en noninvasiv intratrakeal intubation metode, der kan direkte levere eksogene materialer muselunger præsenteres. Denne teknik blev anvendt til at administrere bleomycin til muselunger som en model til at studere lungefibrose. &lt;/ P&gt;

I første omgang blev grøn-farvet løsning, der bruges til at intubere mus til praksis. Lungerne blev reseceret umiddelbart efter intubation at undersøge, hvordan jævnt farve blev fordelt i lunge (figur 2). Denne teknik blev anvendt til at studere bleomycin-induceret lungefibrose hjælp C57BL / 6 mus. Når musene blev intratrachealt intuberet med 1,2 U / kg af bleomycin eller saltvand som kontrol og obduceres 3 uger senere udviklede musene bleomycin-induceret fibrose hele deres lunger, støttet af hi...

Watch this Scientific Journal Video about Noninvasive Intratracheal Intubation to Study the Pathology and Physiology of Mouse Lung at JoVE.com

Noninvasive Intratracheal Intubation to Study the Pathology and Physiology of Mouse Lung (Video) | JoVE

Anvendelsen af ​​en model, der efterligner tilstanden af ​​lungesygdomme hos mennesker er afgørende for at studere patofysiologi og / eller ætiologi af en bestemt sygdom, og for udvikling af terapeutisk intervention. Her en noninvasiv intratrakeal intubation metode, der kan direkte levere eksogene materialer muselunger præsenteres.

Noninvasiv Intratracheal Intubation til at studere Patologi og fysiologi Mouse Lung

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Noninvasive Intratracheal Intubation to Study the Pathology and Physiology of Mouse Lung

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