Name: a doxorubicin-induced cardiomyopathy model in adult zebrafish
Uploaded: 2018-06-07T13:30:00
Description: Watch this Scientific Journal Video about A Doxorubicin-induced Cardiomyopathy Model in Adult Zebrafish at JoVE.com

Dette arbejde blev støttet i en del af en videnskabsmand udvikling tilskud fra American Heart Association (14SDG18160021 til YD), amerikanske NIH R01 tilskud HL 81753 og HL 107304 til XX og XX Mayo instituttet.

Alle procedurer, der beskrives her blev udført i overensstemmelse med retningslinjer for pleje og anvendelse af forsøgsdyr (nationale akademier pressen. 2011), og de blev godkendt af Mayo Clinic institutionelle Animal Care og brug udvalget.
1. voksen zebrafisk forberedelse
<ol><li>Oprette tilstrækkelige ynglende par i krydser tanke at erhverve mindst dobbelt så mange som den samlede fisk brug for DOX injektion. Hvis man sammenligner fisk med forskellige genetiske baggrunde, avle alle fis...

<ol>
	<li>Octavia, 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=Doxorubicin-induced+cardiomyopathy:+from+molecular+mechanisms+to+therapeutic+strategies.">Doxorubicin-induced cardiomyopathy: from molecular mechanisms to therapeutic strategies.</a> J Mol Cell Cardiol. 52 (6), 1213-1225 (2012).</li><li>Singal, P. K., Iliskovic, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doxorubicin-induced+cardiomyopathy.">Doxorubicin-induced cardiomyopathy.</a> N Engl J Med. 339 (13), 900-905 (1998).</li><li>Angsutararux, P., Luanpitpong, S., Issaragrisil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemotherapy-Induced+Cardiotoxicity:+Overview+of+the+Roles+of+Oxidative+Stress.">Chemotherapy-Induced Cardiotoxicity: Overview of the Roles of Oxidative Stress.</a> Oxid Med Cell Longev. , 795602 (2015).</li><li>Ichikawa, 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=Cardiotoxicity+of+doxorubicin+is+mediated+through+mitochondrial+iron+accumulation.">Cardiotoxicity of doxorubicin is mediated through mitochondrial iron accumulation.</a> J Clin Invest. 124 (2), 617-630 (2014).</li><li>Zhang, Y. W., Shi, J., Li, Y. J., Wei, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiomyocyte+death+in+doxorubicin-induced+cardiotoxicity.">Cardiomyocyte death in doxorubicin-induced cardiotoxicity.</a> Arch Immunol Ther Exp (Warsz). 57 (6), 435-445 (2009).</li><li>Sawyer, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anthracyclines+and+heart+failure.">Anthracyclines and heart failure.</a> N Engl J Med. 368 (12), 1154-1156 (2013).</li><li>Zhang, S., 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=Identification+of+the+molecular+basis+of+doxorubicin-induced+cardiotoxicity.">Identification of the molecular basis of doxorubicin-induced cardiotoxicity.</a> Nat Med. 18 (11), 1639-1642 (2012).</li><li>Dodd, D. 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=Doxorubicin+cardiomyopathy+is+associated+with+a+decrease+in+calcium+release+channel+of+the+sarcoplasmic+reticulum+in+a+chronic+rabbit+model.">Doxorubicin cardiomyopathy is associated with a decrease in calcium release channel of the sarcoplasmic reticulum in a chronic rabbit model.</a> J Clin Invest. 91 (4), 1697-1705 (1993).</li><li>Mitry, M. A., Edwards, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doxorubicin+induced+heart+failure:+Phenotype+and+molecular+mechanisms.">Doxorubicin induced heart failure: Phenotype and molecular mechanisms.</a> Int J Cardiol Heart Vasc. 10, 17-24 (2016).</li><li>Aminkeng, F., 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=A+coding+variant+in+RARG+confers+susceptibility+to+anthracycline-induced+cardiotoxicity+in+childhood+cancer.">A coding variant in RARG confers susceptibility to anthracycline-induced cardiotoxicity in childhood cancer.</a> Nat Genet. 47 (9), 1079-1084 (2015).</li><li>Deng, S., 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=Dystrophin-deficiency+increases+the+susceptibility+to+doxorubicin-induced+cardiotoxicity.">Dystrophin-deficiency increases the susceptibility to doxorubicin-induced cardiotoxicity.</a> Eur J Heart Fail. 9 (10), 986-994 (2007).</li><li>Leong, S. L., Chaiyakunapruk, N., Lee, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Candidate+Gene+Association+Studies+of+Anthracycline-induced+Cardiotoxicity:+A+Systematic+Review+and+Meta-analysis.">Candidate Gene Association Studies of Anthracycline-induced Cardiotoxicity: A Systematic Review and Meta-analysis.</a> Sci Rep. 7 (1), 39 (2017).</li><li>Wasielewski, 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=Potential+genetic+predisposition+for+anthracycline-associated+cardiomyopathy+in+families+with+dilated+cardiomyopathy.">Potential genetic predisposition for anthracycline-associated cardiomyopathy in families with dilated cardiomyopathy.</a> Open Heart. 1 (1), e000116 (2014).</li><li>Lebrecht, 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=Dexrazoxane+prevents+doxorubicin-induced+long-term+cardiotoxicity+and+protects+myocardial+mitochondria+from+genetic+and+functional+lesions+in+rats.">Dexrazoxane prevents doxorubicin-induced long-term cardiotoxicity and protects myocardial mitochondria from genetic and functional lesions in rats.</a> Br J Pharmacol. 151 (6), 771-778 (2007).</li><li>QuanJun, 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=Protective+Effects+of+Dexrazoxane+against+Doxorubicin-Induced+Cardiotoxicity:+A+Metabolomic+Study.">Protective Effects of Dexrazoxane against Doxorubicin-Induced Cardiotoxicity: A Metabolomic Study.</a> PLoS One. 12 (1), e0169567 (2017).</li><li>Seifert, C. F., Nesser, M. E., Thompson, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dexrazoxane+in+the+prevention+of+doxorubicin-induced+cardiotoxicity.">Dexrazoxane in the prevention of doxorubicin-induced cardiotoxicity.</a> Ann Pharmacother. 28 (9), 1063-1072 (1994).</li><li>Adams, J. 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=Enhanced+Galphaq+signaling:+a+common+pathway+mediates+cardiac+hypertrophy+and+apoptotic+heart+failure.">Enhanced Galphaq signaling: a common pathway mediates cardiac hypertrophy and apoptotic heart failure.</a> Proc Natl Acad Sci U S A. 95 (17), 10140-10145 (1998).</li><li>Bowles, N. E., Bowles, K. R., Towbin, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+&amp;#34;final+common+pathway&amp;#34;+hypothesis+and+inherited+cardiovascular+disease.+The+role+of+cytoskeletal+proteins+in+dilated+cardiomyopathy.">The &amp;#34;final common pathway&amp;#34; hypothesis and inherited cardiovascular disease. The role of cytoskeletal proteins in dilated cardiomyopathy.</a> Herz. 25 (3), 168-175 (2000).</li><li>Kroumpouzou, E., 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=Common+pathways+for+primary+hypertrophic+and+dilated+cardiomyopathy.">Common pathways for primary hypertrophic and dilated cardiomyopathy.</a> Hybrid Hybridomics. 22 (1), 41-45 (2003).</li><li>Towbin, J. A., Bowles, K. R., Bowles, N. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Etiologies+of+cardiomyopathy+and+heart+failure.">Etiologies of cardiomyopathy and heart failure.</a> Nat Med. 5 (3), 266-267 (1999).</li><li>Liu, 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=Visnagin+protects+against+doxorubicin-induced+cardiomyopathy+through+modulation+of+mitochondrial+malate+dehydrogenase.">Visnagin protects against doxorubicin-induced cardiomyopathy through modulation of mitochondrial malate dehydrogenase.</a> Sci Transl Med. 6 (266), 266ra170 (2014).</li><li>Asimaki, 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=Identification+of+a+new+modulator+of+the+intercalated+disc+in+a+zebrafish+model+of+arrhythmogenic+cardiomyopathy.">Identification of a new modulator of the intercalated disc in a zebrafish model of arrhythmogenic cardiomyopathy.</a> Sci Transl Med. 6 (240), 240ra274 (2014).</li><li>Ding, 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=Haploinsufficiency+of+target+of+rapamycin+attenuates+cardiomyopathies+in+adult+zebrafish.">Haploinsufficiency of target of rapamycin attenuates cardiomyopathies in adult zebrafish.</a> Circ Res. 109 (6), 658-669 (2011).</li><li>Sun, X., 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=Cardiac+hypertrophy+involves+both+myocyte+hypertrophy+and+hyperplasia+in+anemic+zebrafish.">Cardiac hypertrophy involves both myocyte hypertrophy and hyperplasia in anemic zebrafish.</a> PLoS One. 4 (8), e6596 (2009).</li><li>Sun, 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=Activation+of+the+Nkx2.5-Calr-p53+signaling+pathway+by+hyperglycemia+induces+cardiac+remodeling+and+dysfunction+in+adult+zebrafish.">Activation of the Nkx2.5-Calr-p53 signaling pathway by hyperglycemia induces cardiac remodeling and dysfunction in adult zebrafish.</a> Dis Model Mech. 10 (10), 1217-1227 (2017).</li><li>Yang, J., Shah, S., Olson, T. M., Xu, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+GATAD1-Associated+Dilated+Cardiomyopathy+in+Adult+Zebrafish.">Modeling GATAD1-Associated Dilated Cardiomyopathy in Adult Zebrafish.</a> J Cardiovasc Dev Dis. 3 (1), (2016).</li><li>Ding, 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=Trapping+cardiac+recessive+mutants+via+expression-based+insertional+mutagenesis+screening.">Trapping cardiac recessive mutants via expression-based insertional mutagenesis screening.</a> Circ Res. 112 (4), 606-617 (2013).</li><li>Ding, 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=A+modifier+screen+identifies+DNAJB6+as+a+cardiomyopathy+susceptibility+gene.">A modifier screen identifies DNAJB6 as a cardiomyopathy susceptibility gene.</a> JCI Insight. 2 (8), (2017).</li><li>Kinkel, M. D., Eames, S. C., Philipson, L. H., Prince, V. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraperitoneal+injection+into+adult+zebrafish.">Intraperitoneal injection into adult zebrafish.</a> J Vis Exp. (42), (2010).</li><li>Wang, L. 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=Standardized+echocardiographic+assessment+of+cardiac+function+in+normal+adult+zebrafish+and+heart+disease+models.">Standardized echocardiographic assessment of cardiac function in normal adult zebrafish and heart disease models.</a> Dis Model Mech. 10 (1), 63-76 (2017).</li><li>Desai, V. G., 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=Development+of+doxorubicin-induced+chronic+cardiotoxicity+in+the+B6C3F1+mouse+model.">Development of doxorubicin-induced chronic cardiotoxicity in the B6C3F1 mouse model.</a> Toxicol Appl Pharmacol. 266 (1), 109-121 (2013).</li><li>Zhu, W., Shou, W., Payne, R. M., Caldwell, R., Field, 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+mouse+model+for+juvenile+doxorubicin-induced+cardiac+dysfunction.">A mouse model for juvenile doxorubicin-induced cardiac dysfunction.</a> Pediatr Res. 64 (5), 488-494 (2008).</li><li>Chatterjee, K., Zhang, J., Honbo, N., Karliner, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doxorubicin+cardiomyopathy.">Doxorubicin cardiomyopathy.</a> Cardiology. 115 (2), 155-162 (2010).</li><li>Bang, 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=Cardiac+fibroblast-derived+microRNA+passenger+strand-enriched+exosomes+mediate+cardiomyocyte+hypertrophy.">Cardiac fibroblast-derived microRNA passenger strand-enriched exosomes mediate cardiomyocyte hypertrophy.</a> J Clin Invest. 124 (5), 2136-2146 (2014).</li><li>Rassaf, T., Kelm, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protection+from+diabetic+cardiomyopathy+-+putative+role+of+the+retinoid+receptor-mediated+signaling.">Protection from diabetic cardiomyopathy - putative role of the retinoid receptor-mediated signaling.</a> J Mol Cell Cardiol. 59, 179-180 (2013).</li><li>Wahbi, K., 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=Dilated+cardiomyopathy+in+patients+with+mutations+in+anoctamin+5.">Dilated cardiomyopathy in patients with mutations in anoctamin 5.</a> Int J Cardiol. 168 (1), 76-79 (2013).</li><li>Zhou, M. D., Sucov, H. M., Evans, R. M., Chien, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinoid-dependent+pathways+suppress+myocardial+cell+hypertrophy.">Retinoid-dependent pathways suppress myocardial cell hypertrophy.</a> Proc Natl Acad Sci U S A. 92 (16), 7391-7395 (1995).</li><li>Hershman, D. 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=Doxorubicin,+cardiac+risk+factors,+and+cardiac+toxicity+in+elderly+patients+with+diffuse+B-cell+non-Hodgkin's+lymphoma.">Doxorubicin, cardiac risk factors, and cardiac toxicity in elderly patients with diffuse B-cell non-Hodgkin's lymphoma.</a> J Clin Oncol. 26 (19), 3159-3165 (2008).</li><li>Silber, J. H., Barber, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doxorubicin-induced+cardiotoxicity.">Doxorubicin-induced cardiotoxicity.</a> N Engl J Med. 333 (20), 1359-1360 (1995).</li><li>Von Hoff, D. 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=Risk+factors+for+doxorubicin-induced+congestive+heart+failure.">Risk factors for doxorubicin-induced congestive heart failure.</a> Ann Intern Med. 91 (5), 710-717 (1979).</li><li>Pugach, E. K., Li, P., White, R., Zon, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retro-orbital+injection+in+adult+zebrafish.">Retro-orbital injection in adult zebrafish.</a> J Vis Exp. (34), (2009).</li><li>Zang, L., Morikane, D., Shimada, Y., Tanaka, T., Nishimura, N. <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+protocol+for+the+oral+administration+of+test+chemicals+to+adult+zebrafish.">A novel protocol for the oral administration of test chemicals to adult zebrafish.</a> Zebrafish. 8 (4), 203-210 (2011).</li><li>Collymore, C., Rasmussen, S., Tolwani, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gavaging+adult+zebrafish.">Gavaging adult zebrafish.</a> J Vis Exp. (78), (2013).</li></ol>

For at modellere en progressiv DIC, var dosis på 20 mg/kg DOX bestemmes eksperimentelt som den højeste dosis, der ikke medfører væsentlige fisk død under 1 wpi men stadig resulterer i fisk død og reduktion af hjertets funktion efter 4 wpi (figur 3 og Figur 4 c). Denne dosis kan sammenlignes til dem, der ofte anvendes i gnavere DIC modeller (15-25 mg/kg) og den grænse kumulative dosis hos mennesker (550 mg/m2, hvilket svarer til 15 mg/kg)<sup c...

Doxorubicin (DOX), også kaldet Adriamycin, er blevet udviklet som en anti-neoplastiske medicin siden 1960s1,2. Det er nu endnu bruges aktivt som en vigtig kemoterapeutiske stof for et bredt spektrum af tumorer. Men kliniske anvendelse af DOX var hæmmet af dens dosisafhængig toksicitet, især cardiotoxicity karakteriseret ved variabel symptomerne spænder fra asymptomatisk elektrokardiografiske ændringer til pericarditis og decompensated kardiomyopati 1 , 2. indtil mindst tre store hypoteser er blevet rejst for at forklare DIC, herunder aktiveret reaktive ilt arter (ROS)1,3,4,5, hæmning af topoisomerase II-β ( TOP2β)6,7, og graduering af intracellulære calcium frigive1,8,9. Akkumulere beviser foreslår også genetisk disposition som en central risikofaktor for DIC10,11,12,13. Gen identiteter relateret til disse DIC prædisposition, men stadig stort set ukendt. Save er det eneste adjuverende agent godkendt af US Food and Drug Administration (FDA) til behandling af DIC, men med begrænset gennemførelsen14,15,16, understreger behovet for at identificere yderligere terapeutiske strategier. Dyremodeller af DIC er derfor undersøgt for disse formål. På grund af deres tilgængelighed og enkelhed, Mekanistiske undersøgelser på DIC modeller kunne potentielt har bredere virkninger på andre typer af cardiomyopatier: fælles patogenese kan deles mellem cardiomyopatier af forskellige etiologies, især på senere patologisk faser17,18,19,20.
Ud over gnavere modeller af DIC, er zebrafisk DIC modeller med højere overførselshastighed blevet udviklet for at lette opdagelsen af nye genetiske faktorer og therapeutics. En embryonale DIC model har været etableret i gennemsigtig zebrafisk embryoner til screening terapeutiske forbindelser21. Cardiomyopatier er voksen indsættende sygdomme med en progressiv patogenese, udviklet voksen zebrafisk kardiomyopati modeller har været22,23,24,25,26. Vi genereres den første erhvervede model for kardiomyopati som følge af kronisk anæmi24, efterfulgt af DIC som den anden erhvervede kardiomyopati model i voksen zebrafisk23. Vi fandt, at injektion af en enkelt bolus af DOX i voksen zebrafisk inducerer cardiotoxicity, der består af en akut fase groft inden for 1 uge efter injektion (wpi), efterfulgt af en kronisk fase af kardiomyopati til 6 måneder efter injektion. Mens haploinsufficiency til mekanistiske målet på rapamycin(mtor) forbedringer kardiomyopati i den kroniske fase, overdriver det fiskedødelighed på den akutte fase, hvilket understreger værdien af den voksne DIC model at skelne fase-afhængige mekanismer23. Vi viste yderligere, at den voksne DIC model kan bruges til at understrege en samling af zebrafisk pattedyrsceller hjerte (ZIC) mutanter, der bliver genereret via en transposon-baserede pattedyrsceller mutagenese tilgang27. En pilot skærm identificeret 3 kendte kardiomyopati gener samt DnaJ (Hsp40) homolog, underfamilie B og medlem 6b (dnajb6b) som ny DIC modtagelighed gener28. Derfor, generation af voksen DIC model i zebrafisk førte til en ny metode, der systematisk muliggør identifikation af genetisk modificering af DIC, som supplerer de eksisterende genome-wide association studier (GWAS) og kvantitative træk locus (QTL ) analyse.
Under den generation og gennemførelse af voksen zebrafisk DIC model bemærkede vi betydelige variationer mellem forskellige forskere og/eller selv blandt forskellige injektioner udført af den samme investigator. Langsgående arten af modellen stiller udfordringer til at registrere resultaterne fra forskellige efterforskere og den sekventielle fejlfindingsprocessen. For at lette anvendelsen af denne enkle kardiomyopati-inducerende stress metode af Fællesskabets forskning, vi beskriver vores protokol i detaljer, findes to typer IP injektion, og diskutere overvejelser for at reducere variationer mellem forskellige forskere.

<table>
	<tbody>
		<tr>
			<td>Crossing tank</td>
			<td>Aquaneering</td>
			<td>ZHCT100</td>
			<td>Fish breeding</td>
		</tr>
		<tr>
			<td>Incubator</td>
			<td>ThermoFisher</td>
			<td></td>
			<td>Maintaining embryo</td>
		</tr>
		<tr>
			<td>3 L medium tank</td>
			<td>Aquaneering</td>
			<td>ZT280</td>
			<td>Maintaining fish</td>
		</tr>
		<tr>
			<td>Paramecia</td>
			<td>Carolina</td>
			<td>131560</td>
			<td>Food for juvenile fish</td>
		</tr>
		<tr>
			<td>Live hatched brine shrimp</td>
			<td>in house</td>
			<td></td>
			<td>Food for adult fish</td>
		</tr>
		<tr>
			<td>Doxorubicin hydrochloride</td>
			<td>Sigma</td>
			<td>D1515-10MG</td>
			<td></td>
		</tr>
		<tr>
			<td>1.5 ml safe-lock tube</td>
			<td>Eppendorf</td>
			<td>No. 022363204</td>
			<td>For drug storage</td>
		</tr>
		<tr>
			<td>Aluminum foil paper</td>
			<td>Fisher</td>
			<td>1213104</td>
			<td>For preventing light exposure</td>
		</tr>
		<tr>
			<td>Proteinase K</td>
			<td>Roche</td>
			<td>No. 03115887001</td>
			<td>For dechorionating embryo</td>
		</tr>
		<tr>
			<td>Hank&#39;s balanced salt solution (HBBS)</td>
			<td>ThermoFisher</td>
			<td>14025076</td>
			<td>Vehicle for DOX</td>
		</tr>
		<tr>
			<td>100 mm petri dish</td>
			<td>Falcon</td>
			<td>431741</td>
			<td></td>
		</tr>
		<tr>
			<td>10 &#956;L NanoFil micro-syringe</td>
			<td>WPI</td>
			<td>NANOFIL</td>
			<td>For injection</td>
		</tr>
		<tr>
			<td>34 gauge needle&#160;</td>
			<td>WPI</td>
			<td>NF34BV-2</td>
			<td>For injection</td>
		</tr>
		<tr>
			<td>Tricaine</td>
			<td>Argent</td>
			<td>MS-222</td>
			<td>Anesthetizing fish</td>
		</tr>
		<tr>
			<td>96 well plate</td>
			<td>Costar</td>
			<td>3539</td>
			<td>For embryo drug treatment</td>
		</tr>
		<tr>
			<td>Transfer pipette</td>
			<td>Bel-art product</td>
			<td>F37898</td>
			<td>For transfering embryo</td>
		</tr>
	</tbody>
</table>

a doxorubicin-induced cardiomyopathy model in adult zebrafish

Den genetisk tilgængelige voksen zebrafisk (Danio rerio) er blevet mere og mere brugt som en hvirveldyr model for at forstå menneskers sygdomme såsom kardiomyopati. På grund af dens bekvemmelighed og imødekommenhed til høj overførselshastighed genetiske manipulationer, åbner generation af erhvervede kardiomyopati modeller, såsom doxorubicin-induceret kardiomyopati (DIC) model i voksen zebrafisk, døre til nye forskning veje, herunder at opdage kardiomyopati modifikatorer via fremad genetisk screening. Forskellig fra den embryonale zebrafisk DIC model, både indledende akut og senere kronisk faser af kardiomyopati kan bestemmes i den voksne zebrafisk DIC model, giver undersøgelsen etape-afhængige signaling mekanismer og terapeutiske strategier. Dog kan varierende resultater opnås med den nuværende model, selv i hænderne på erfarne efterforskere. For at lette fremtidig gennemførelse af DIC model, præsenterer vi en detaljeret protokol om hvordan man kan generere denne DIC model i voksen zebrafisk og beskrive to alternative måder at intraperitoneal (IP) injektion. Vi diskutere yderligere muligheder at reducere variationer for at få pålidelige resultater og stille forslag om, hvordan man korrekt fortolke resultaterne.

Her, præsenteres to metoder til at udføre IP injektion til model DIC i voksen zebrafisk. Mens du bruger den klassiske, etablerede IP injektion metode29, blev det bemærket, at den injicerede DOX løsning (rød farve) undertiden kunne sive ud fra den placering, hvor nålen gennemtrænges. Den alternative IP injektion bruger en anden placering for nål penetration, der er 3-4 mm fra bughinden hvor DOX er frigivet (figur 1A), der effekt...

Watch this Scientific Journal Video about A Doxorubicin-induced Cardiomyopathy Model in Adult Zebrafish at JoVE.com

A Doxorubicin-induced Cardiomyopathy Model in Adult Zebrafish

En Doxorubicin-induceret kardiomyopati Model i voksen zebrafisk

En metode til at generere en doxorubicin-induceret kardiomyopati model i voksen zebrafisk (Danio rerio) er beskrevet her. To alternative måder at intraperitoneal injektion præsenteres og betingelser, der skal reducere variationer mellem forskellige eksperimentelle grupper diskuteres.

The genetically accessible adult zebrafish (Danio rerio) has been increasingly used as a vertebrate model for understanding human diseases such as ...

Research

JoVE Journal

Medicine

Induction of Myocardial Infarction in Adult Zebrafish Using Cryoinjury

Induktion af myokardieinfarkt i Adult Zebrafisk Brug kryobeskadigelse

The mammalian heart is incapable of significant regeneration following an acute injury such as myocardial infarction1. By contrast, urodele amphibians and ...

Assessing Teratogenic Changes in a Zebrafish Model of Fetal Alcohol Exposure

Evaluering Teratogene Ændringer i en Zebrafisk Model af Føtalt Alkohol Eksponering

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, ...

Screening for Melanoma Modifiers using a Zebrafish Autochthonous Tumor Model

Screening for Melanoma Modifiers anvendelse af et zebrafisk Autokton tumormodel

Genomic studies of  human cancers have yielded a wealth of information about genes that are altered  in tumors1,2,3. A challenge arising from these ...

A Zebrafish Model of Diabetes Mellitus and Metabolic Memory

En zebrafisk Model af diabetes mellitus og Metabolic Memory

Diabetes mellitus currently affects 346 million individuals and this is projected to increase to 400 million by 2030. Evidence from both the laboratory ...

A Possible Zebrafish Model of Polycystic Kidney Disease: Knockdown of wnt5a Causes Cysts in Zebrafish Kidneys

A Possible Zebrafish Model of Polycystic Kidney Disease: Knockdown of wnt5a Causes Cysts in Zebrafish Kidneys

En mulig Zebrafisk Model af polycystisk nyresygdom: Knockdown af<em&gt; Wnt5a</em&gt; Årsager cyster i zebrafisk Nyrer

Polycystic kidney disease (PKD) is one of the most common causes of end-stage kidney disease, a devastating disease for which there is no cure. The ...

Studying Diabetes Through the Eyes of a Fish: Microdissection, Visualization, and Analysis of the Adult tg(fli:EGFP) Zebrafish Retinal Vasculature

Studying Diabetes Through the Eyes of a Fish: Microdissection, Visualization, and Analysis of the Adult tgfli:EGFP Zebrafish Retinal Vasculature

At studere Diabetes gennem øjnene på en fisk: Microdissection, visualisering og analyse af de voksne tg(fli:EGFP) zebrafisk Retinal Vaskulaturen

Diabetic retinopathy is the leading cause of blindness among middle-aged adults. The rising prevalence of diabetes worldwide will make the prevention of ...

Tachycardia-Induced Cardiomyopathy As a Chronic Heart Failure Model in Swine

Takykardi-induceret kardiomyopati som kronisk hjertesvigt Model hos svin

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

Isometrisk kontraktilitet måling af musen mesenterica arterie ved hjælp af Wire blodkar

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Isolation of Atrial Myocytes from Adult Mice

Isolering af atrial Myocytes fra voksne mus

The electrophysiological properties of atrial myocytes importantly affect overall cardiac function. Alterations in the underlying ionic currents ...

Murine Myocardial Infarction Model using Permanent Ligation of Left Anterior Descending Coronary Artery

Murine myokardieinfarkt model ved hjælp af permanent ligation af venstre anterior faldende koronararterie

Myocardial infarction (MI) and acute coronary diseases are among the most prominent causes of death in population with western lifestyle. The murine ...