Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction

Virag, Jitka A. I.; Lust, Robert M.

doi:10.3791/2581

Cited by 39 publications

(27 citation statements)

References 64 publications

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“…Acute MI was induced as previously described [27]. Briefly, mice were anesthetized with 2% isoflurane, intubated endotracheally with a 22 gauge intravenous catheter and placed in a supine position on the warming surgery platform.…”

Section: Methodsmentioning

confidence: 99%

Hiding inside? Intracellular expression of non-glycosylated c-kit protein in cardiac progenitor cells

et al. 2016

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Cardiac progenitor cells including c-kit+ cells and Cardiosphere-derived cells (CDCs) play important roles in cardiac repair and regeneration. CDCs were reported to contain only small subpopulations of c-kit+ cells and recent publications suggested that depletion of the c-kit+ subpopulation of cells has no effect on regenerative properties of CDCs. However, our current study showed that the vast majority of CDCs from murine heart actually express c-kit, albeit, in an intracellular and non-glycosylated form. Immunostaining and flow cytometry showed that the fluorescent signal indicative of c-kit immunstaining significantly increased when cell membranes were permeabilized. Western blots further demonstrated that glycosylation of c-kit was increased during endothelial differentiation in a time dependent manner. Glycosylation inhibition by 1-deoxymannojirimycin hydrochloride (1-DMM) blocked c-kit glycosylation and reduced expression of endothelial cell markers such as Flk-1 and CD31 during differentiation. Pretreatment of these cells with a c-kit kinase inhibitor (imatinib mesylate) also attenuated Flk-1 and CD31 expression. These results suggest that c-kit glycosylation and its kinase activity are likely needed for these cells to differentiate into an endothelial lineage. In vivo, we found that intracellular c-kit expressing cells are located in the wall of cardiac blood vessels in mice subjected to myocardial infarction. In summary, our work demonstrated for the first time that c-kit is not only expressed in CDCs but may also directly participate in CDC differentiation into an endothelial lineage.

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Section: Methodsmentioning

confidence: 99%

Hiding inside? Intracellular expression of non-glycosylated c-kit protein in cardiac progenitor cells

et al. 2016

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“…Several laboratories have introduced various methods to establish murine myocardial I/R models [28, 29]. Among these methods, the classic method has been widely accepted and applied [29].…”

Section: Discussionmentioning

confidence: 99%

“…Among these methods, the classic method has been widely accepted and applied [29]. In addition, this method can induce myocardial I/R using a standard approach; however, during the operation period, the chest cavity must be re-opened to loosen the slipknot to perform reperfusion.…”

Section: Discussionmentioning

confidence: 99%

Outer Balloon Ligation Increases Success Rate of Ischemia-Reperfusion Injury Model in Mice

Zhai

Gao

et al. 2016

PLoS ONE

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BackgroundCoronary artery disease is a growing public health problem and a major cause of morbidity and mortality. Experimental animal models provide valuable tools for studying myocardial ischemia reperfusion (I/R) injury in vivo.ObjectiveThe purpose of this study was to describe a new method (outer balloon ligation) to induce myocardial I/R injury in mice.MethodsNinety-nine male C57BL/6J mice were randomly divided into three groups: sham group, classic method group (I/R-C) and the new method group (I/R-N). The surgical procedure and recovery time were recorded. The levels of TNF-α, IL-6, cTnT and LDH were detected by ELISA kits. Hematoxylin-eosin staining was applied to assess neutrophil infiltration. Moreover, surgical survival, myocardial infarction areas, and cardiac function measurements were also recorded.ResultsThe reperfusion operation time in the I/R-N group were markedly less than the I/R-C group (14.73±2.86 vs. 168.60±33.01 sec, p <0.0001). Similarly, the recovery time in I/R-N group was shorter than the I/R-C group (45.39±15.39 vs. 101.70±19.33 min, p <0.0001). The levels of TNF-α and IL-6 in I/R-N group were also markedly lower than in I/R-C group (136.5±22.21 vs. 170.5±24.79 pg/ml, p <0.05 and 100.3±23.74 vs. 144.40±22.24 pg/ml, p <0.001). Compared I/R-N group with I/R-C group, the levels of neutrophil infiltration, cTnT and LDH had no significant differences. Surgical survival rate was 96.7% in the I/R-N group, which was significantly improved compared to the rate of 80% in the I/R-C group. However, there were no significant differences in the areas of myocardial infarction and cardiac function between the two groups.ConclusionsCompared with the classic method, our new method of inducing myocardial I/R injury has higher efficiency and less tissue damage in mice, but achieves the same modeling effects.

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“…Arrest the hearts in diastole and fix with formalin. Use the procedure of the heart harvesting that has been described previously 22 . 11.…”

Section: Methodsmentioning

confidence: 99%

Ultrasound Based Assessment of Coronary Artery Flow and Coronary Flow Reserve Using the Pressure Overload Model in Mice

Chang

Fisch

Chen

et al. 2015

JoVE

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Transthoracic Doppler echocardiography (TTDE) is a clinically useful, noninvasive tool for studying coronary artery flow velocity and coronary flow reserve (CFR) in humans. Reduced CFR is accompanied by marked intramyocardial and pericoronary fibrosis and is used as an indication of the severity of dysfunction. This study explores, step-by-step, the real-time changes measured in the coronary flow velocity, CFR and systolic to diastolic peak velocity (S/D) ratio in the setting of an aortic banding model in mice. By using a Doppler transthoracic imaging technique that yields reproducible and reliable data, the method assesses changes in flow in the septal coronary artery (SCA), for a period of over two weeks in mice, that previously either underwent aortic banding or thoracotomy.During imaging, hyperemia in all mice was induced by isoflurane, an anesthetic that increased coronary flow velocity when compared with resting flow. All images were acquired by a single imager. Two ratios, (1) CFR, the ratio between hyperemic and baseline flow velocities, and (2) systolic (S) to diastolic (D) flow were determined, using a proprietary software and by two independent observers. Importantly, the observed changes in coronary flow preceded LV dysfunction as evidenced by normal LV mass and fractional shortening (FS).The method was benchmarked against the current gold standard of coronary assessment, histopathology. The latter technique showed clear pathologic changes in the coronary artery in the form of peri-coronary fibrosis that correlated to the flow changes as assessed by echocardiography.The study underscores the value of using a non-invasive technique to monitor coronary circulation in mouse hearts. The method minimizes redundant use of research animals and demonstrates that advanced ultrasound-based indices, such as CFR and S/D ratios, can serve as viable diagnostic tools in a variety of investigational protocols including drug studies and the study of genetically modified strains. Video LinkThe video component of this article can be found at

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Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction

Cited by 39 publications

References 64 publications

Hiding inside? Intracellular expression of non-glycosylated c-kit protein in cardiac progenitor cells

Hiding inside? Intracellular expression of non-glycosylated c-kit protein in cardiac progenitor cells

Outer Balloon Ligation Increases Success Rate of Ischemia-Reperfusion Injury Model in Mice

Ultrasound Based Assessment of Coronary Artery Flow and Coronary Flow Reserve Using the Pressure Overload Model in Mice

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