Name: dissection of the mouse pancreas for histological analysis and metabolic profiling
Uploaded: 2017-08-19T14:00:00
Description: Watch this Scientific Journal Video about Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling at JoVE.com

MAK anerkender støtte til dette arbejde fra National Institutes of Health / National Cancer Institute giver nummer - 1R15CA152985-01A1. Dette projekt er også blevet støttet af Miami University Undergraduate Research Award Program, Miami University ph.d.-bachelor muligheder for Scholarship Program og Miami University forskere sommerprogram.

de procedurer, der udføres i videoen og beskrevet nedenfor er blevet godkendt af institutionelle Animal Care og brug udvalg (IACUC) på Miami University.
 1. forberedelse og Stimulus Test <ol><li>etablere to særskilte områder for den kirurgiske procedure, operationsbordet og tabellen efter operationen. Stage begge områder med alle materialer og redskaber nødvendigt.
	<ol><li>Fase de opererer tabel under en ventileret hætte. Arrangere tabellen med udstyret på en måde, der giver mulig...

<ol>
	<li>Fesinmeyer, M. D., Austin, M. A., Li, C. I., De Roos, A. J., Bowen, 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=Differences+in+Survival+by+Histologic+Type+of+Pancreatic+Cancer.">Differences in Survival by Histologic Type of Pancreatic Cancer.</a> Cancer Epidemiol Biomarkers Prev. 14 (7), 1766-1773 (2005).</li><li>Jackson-Grusby, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+cancer+in+mice.">Modeling cancer in mice.</a> Oncogene. 21 (35), 5504-5514 (2002).</li><li>Heid, I., 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=Early+requirement+of+Rac1+in+a+mouse+model+of+pancreatic+cancer.">Early requirement of Rac1 in a mouse model of pancreatic cancer.</a> Gastroenterology. 141 (2), 719-730 (2011).</li><li>Hingorani, S. 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=Preinvasive+and+invasive+ductal+pancreatic+cancer+and+its+early+detection+in+the+mouse.">Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse.</a> Cancer cell. 4 (6), 437-450 (2003).</li><li>Shi, 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=KRAS2+Mutations+in+Human+Pancreatic+Acinar-Ductal+Metaplastic+Lesions+Are+Limited+to+Those+with+PanIN+Implications+for+the+Human+Pancreatic+Cancer+Cell+of+Origin.">KRAS2 Mutations in Human Pancreatic Acinar-Ductal Metaplastic Lesions Are Limited to Those with PanIN Implications for the Human Pancreatic Cancer Cell of Origin.</a> Mol Cancer Res. 7 (2), 230-236 (2009).</li><li>LaConti, J. 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=Distinct+serum+metabolomics+profiles+associated+with+malignant+progression+in+the+KrasG12D+mouse+model+of+pancreatic+ductal+adenocarcinoma.">Distinct serum metabolomics profiles associated with malignant progression in the KrasG12D mouse model of pancreatic ductal adenocarcinoma.</a> BMC Genomics. 16 (1), 1-10 (2015).</li><li>Ludwig, M. 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=Surveying+the+serologic+proteome+in+a+tissue-specific+kras(G12D)+knockin+mouse+model+of+pancreatic+cancer.">Surveying the serologic proteome in a tissue-specific kras(G12D) knockin mouse model of pancreatic cancer.</a> Proteomics. 16 (3), 516-531 (2016).</li><li>Taylor, D. K., Mook, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoflurane+Waste+Anesthetic+Gas+Concentrations+Associated+with+the+Open-Drop+Method.">Isoflurane Waste Anesthetic Gas Concentrations Associated with the Open-Drop Method.</a> J Am Assoc Lab Anim Sci. 48 (1), 61-64 (2009).</li><li>Parasuraman, S., Raveendran, R., Kesavan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+sample+collection+in+small+laboratory+animals.">Blood sample collection in small laboratory animals.</a> J Pharmacol Pharmacother. 1 (2), 87-93 (2010).</li><li>Li, D. S., Yuan, Y. H., Tu, H. J., Liang, Q. L., Dai, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+protocol+for+islet+isolation+from+mouse+pancreas.">A protocol for islet isolation from mouse pancreas.</a> Nature Protocols. 4 (11), 1649-1652 (2009).</li><li>Auer, H., 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+effects+of+frozen+tissue+storage+conditions+on+the+integrity+of+RNA+and+protein.">The effects of frozen tissue storage conditions on the integrity of RNA and protein.</a> Biotech Histochem. 89 (7), 518-528 (2014).</li><li>Ma, J., Leung, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Limbic+System+Participates+in+Mediating+the+Effects+of+General+Anesthetics.">Limbic System Participates in Mediating the Effects of General Anesthetics.</a> Neuropsychopharmacology. 31, 1177-1192 (2006).</li><li>Ijichi, H., 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=Aggressive+pancreatic+ductal+adenocarcinoma+in+mice+caused+by+pancreas-specific+blockade+of+transforming+growth+factor-&#946;+signaling+in+cooperation+with+active+Kras+expression.">Aggressive pancreatic ductal adenocarcinoma in mice caused by pancreas-specific blockade of transforming growth factor-&#946; signaling in cooperation with active Kras expression.</a> Genes Dev. 20 (22), 3147-3160 (2006).</li><li>Abiatari, I., 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=Moesin-dependent+cytoskeleton+remodelling+is+associated+with+an+anaplastic+phenotype+of+pancreatic+cancer.">Moesin-dependent cytoskeleton remodelling is associated with an anaplastic phenotype of pancreatic cancer.</a> J Cell Mol Med. 14 (5), 1166-1179 (2010).</li><li>Goodpaster, A. M., Romick-Rosendale, L. E., Kennedy, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=a+Statistical+significance+analysis+of+nuclear+magnetic+resonance-based+metabonomics+data.">a Statistical significance analysis of nuclear magnetic resonance-based metabonomics data.</a> Analytical biochemistry. 401, 134-143 (2010).</li><li>Yadav, 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=Idiopathic+Tumefactive+Chronic+Pancreatitis:+Clinical+Profile,+Histology,+and+Natural+History+After+Resection.">Idiopathic Tumefactive Chronic Pancreatitis: Clinical Profile, Histology, and Natural History After Resection.</a> Clin Gastroenterol Hepatol. 1 (2), 129-135 (2003).</li></ol>

Betydning med hensyn til eksisterende metoder Mens andre uformelle videoer af musen dissektioner findes, denne video artikel giver den første professionelle kvalitet, peer reviewede, visuel demonstration af alle de detaljerede trin påkrævet for udvinding og høst af musen bugspytkirtlen af dissektion10 . Med bugspytkirtlen er en vigtigste organ for metaboliske aktivitet og insulin produktion, dissektion og høst i bugspytkirtlen giver mulighed for bevarelse af den fysiologis...

Musen har vist sig som en vigtig dyremodel af human bugspytkirtelkræft1,2. I Ptf1acre / +; LSL-KrasG12D / + musemodel, Kristen rotte sarkom (K-Ras) onkogen aktiveres udelukkende i bugspytkirtlen, resulterer i indledningen af præcancerøse læsioner i bugspytkirtlen, kendt som pancreas intraepithelial neoplasias (PanINs), fremskridtet til pancreas duktalt adenocarcinomer, almindeligvis benævnt PDACs3. Denne mus modelsystem giver en af de bedste tilgængelige dyremodeller for humane bugspytkirtelkræft4,5, med den yderligere fordel, at PanINs dukke op inden for de første fem måneder af livet og ofte fremskridt til PDAC inden for en enkelt år4,5, hvorimod kræft i bugspytkirtlen hyppigst opstår hos mennesker 60-70 år.
Udtræk af bugspytkirtlen af dissektion fra Ptf1acre / +; LSL-KrasG12D / + mus på forskellige alderstrin giver mulighed for detaljerede langsgående histologisk undersøgelse af udviklingen af kræft i bugspytkirtlen, lige fra de tidligste PanIN faser gennem progression til PDAC3,4 , 5. høst bugspytkirtlen på aldre spænder fra fem til femten måneder kan også bruges til at forberede væv ekstrakter til at karakterisere de globale forandringer i bugspytkirtlen4 metabolisme, der opstår under overgangen fra sund til sygt væv 6,7.
Denne artikel præsenterer en komplet visuel vejledning af de trin, der kræves for at udføre en mus bugspytkirtlen udvinding og indeholder retningslinjer for opbevaring af en bugspytkirtlen for yderligere analyse. Denne guide vil være lige så værdifuld for personer udfører forskning på andre pancreas sygdomme, herunder type I diabetes, og er især nyttigt til studerende og efterforskere nye til forskning, der indebærer høst mus bugspytkirtlen ved brug af dissektion for metabolisk profilering eller histologiske analyser.

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dissection of the mouse pancreas for histological analysis and metabolic profiling

Vi har undersøgt bugspytkirtlen specifikke transkriptionsfaktor, 1a cre-recombinase; LOX-stop-lox-Kristen rotte sarkom, glycin til aspartinsyre på 12-codon (Ptf1acre / +; LSL-KrasG12D / +) mus stamme som en model af menneskelige bugspytkirtelkræft. Målet med vores aktuelle undersøgelser er at identificere roman metaboliske markører for kræft i bugspytkirtlen progression. Vi har udført, metabolisk profilering af urin, afføring, blod, og bugspytkirtlen væv ekstrakter og histologiske analyser af bugspytkirtlen til at iscenesætte kræft progression. Musen bugspytkirtlen er ikke en veldefineret solid orgel som hos mennesker, men er snarere et diffust fordelt blødt væv, der ikke er let identificeres af enkeltpersoner uvante med musen indre anatomi eller af personer, der har lidt eller ingen erfaring udfører musen orgel dissektioner. Formålet med denne artikel er at give en detaljeret trinvis visuel demonstration at guide nybegyndere i fjernelse af musen bugspytkirtlen ved dissektion. Denne artikel bør være særligt værdifuldt til studerende og efterforskere ny forskning, der kræver høst af musen bugspytkirtlen af dissektion for metabolisk profilering eller histologiske analyser.

Figur 1 viser et overblik over miljøområdet operationelle og figur 2 viser post drift området. Mens denne indstilling giver den minimale mængde af udstyr og iscenesættelse, enkeltpersoner kan vælge at ændre dette passer bedst til individuelle behov. Protokollen skal optimeres efter de specifikke behov i eksperimentet. Denne procedure er udført på en måde, der afslutter livet af en mus, der kræver passende euthanization...

Watch this Scientific Journal Video about Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling at JoVE.com

Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling

Denne video artikel giver en detaljeret demonstration af de procedurer, der kræves for at kunne fjerne bugspytkirtlen fra en mus af dissektion for histologiske analyse og metabolisk profilering.

Dissektion af musen bugspytkirtlen for histologiske analyse og metabolisk profilering

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at ...

Research

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