Left Ventricular Function In Mice Research Articles

HTRA1-driven detachment of type I collagen from endoplasmic reticulum contributes to myocardial fibrosis in dilated cardiomyopathy

BackgroundThe aberrant secretion and excessive deposition of type I collagen (Col1) are important factors in the pathogenesis of myocardial fibrosis in dilated cardiomyopathy (DCM). However, the precise molecular mechanisms underlying the synthesis and secretion of Col1 remain unclear.Methods and resultsRNA-sequencing analysis revealed an increased HtrA serine peptidase 1 (HTRA1) expression in patients with DCM, which is strongly correlated with myocardial fibrosis. Consistent findings were observed in both human and mouse tissues by immunoblotting, quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunohistochemistry, and immunofluorescence analyses. Pearson’s analysis showed a markedly positive correlation between HTRA1 level and myocardial fibrosis indicators, including extracellular volume fraction (ECV), native T1, and late gadolinium enhancement (LGE), in patients with DCM. In vitro experiments showed that the suppression of HTRA1 inhibited the conversion of cardiac fibroblasts into myofibroblasts and decreased Col1 secretion. Further investigations identified the role of HTRA1 in promoting the formation of endoplasmic reticulum (ER) exit sites, which facilitated the transportation of Col1 from the ER to the Golgi apparatus, thereby increasing its secretion. Conversely, HTRA1 knockdown impeded the retention of Col1 in the ER, triggering ER stress and subsequent induction of ER autophagy to degrade misfolded Col1 and maintain ER homeostasis. In vivo experiments using adeno-associated virus-serotype 9-shHTRA1-green fluorescent protein (AAV9-shHTRA1-GFP) showed that HTRA1 knockdown effectively suppressed myocardial fibrosis and improved left ventricular function in mice with DCM.ConclusionsThe findings of this study provide valuable insights regarding the treatment of DCM-associated myocardial fibrosis and highlight the therapeutic potential of targeting HTRA1-mediated collagen secretion.

A RGS7-CaMKII complex drives myocyte-intrinsic and myocyte-extrinsic mechanisms of chemotherapy-induced cardiotoxicity

Dose-limiting cardiotoxicity remains a major limitation in the clinical use of cancer chemotherapeutics. Here, we describe a role for Regulator of G protein Signaling 7 (RGS7) in chemotherapy-dependent heart damage, the demonstration for a functional role of RGS7 outside of the nervous system and retina. Though expressed at low levels basally, we observed robust up-regulation of RGS7 in the human and murine myocardium following chemotherapy exposure. In ventricular cardiomyocytes (VCM), RGS7 forms a complex with Ca2+/calmodulin-dependent protein kinase (CaMKII) supported by key residues (K412 and P391) in the RGS domain of RGS7. In VCM treated with chemotherapeutic drugs, RGS7 facilitates CaMKII oxidation and phosphorylation and CaMKII-dependent oxidative stress, mitochondrial dysfunction, and apoptosis. Cardiac-specific RGS7 knockdown protected the heart against chemotherapy-dependent oxidative stress, fibrosis, and myocyte loss and improved left ventricular function in mice treated with doxorubicin. Conversely, RGS7 overexpression induced fibrosis, reactive oxygen species generation, and cell death in the murine myocardium that were mitigated following CaMKII inhibition. RGS7 also drives production and release of the cardiokine neuregulin-1, which facilitates paracrine communication between VCM and neighboring vascular endothelial cells (EC), a maladaptive mechanism contributing to VCM dysfunction in the failing heart. Importantly, while RGS7 was both necessary and sufficient to facilitate chemotherapy-dependent cytotoxicity in VCM, RGS7 is dispensable for the cancer-killing actions of these same drugs. These selective myocyte-intrinsic and myocyte-extrinsic actions of RGS7 in heart identify RGS7 as an attractive therapeutic target in the mitigation of chemotherapy-driven cardiotoxicity.

The Regulation of Heme Metabolism in Myocytes by Hypoxia and Ischemia: Substrate‐Enzyme Dyssynchrony

Current strategies for the treatment of acute myocardial infarction (AMI) are limited therefore better pharmacological interventions are needed to improve cardiac function post‐injury. Overexpression of the enzyme heme oxygenase‐1 (HMOX1) has been shown to exert cytoprotective effects in models of acute myocardial infarction and represents a highly attractive therapeutic target. HMOX1 is the stress‐inducible enzyme responsible for the catabolism of heme, the functional backbone by which oxygen is carried in proteins like hemoglobin and cytochromes in the electron transport chain. As a potent inducer of HMOX1 and an FDA‐approved heme‐surrogate for the treatment of porphyria, hemin represents a novel treatment strategy for AMI.Translation of HMOX1‐targeted therapeutics to the clinic has been stymied by a singular focus on HMOX1 and an incomplete understanding of fundamental heme metabolism regulation in response to hypoxia, ischemia or pharmacological stimulation with hemin. To date, studies investigating hemin pharmacology and its use in models of ischemic injury are limited. The objective of the present study was to provide a temporal and pharmacokinetic characterization of the changes in heme regulatory enzyme expression in response to hemin and ischemic cardiac injury (i.e. AMI).Echocardiographic, hemodynamic and protein quantification analyses were performed in male CD‐1 mice following a single intraperitoneal hemin injection, permanent ligation AMI, or daily hemin treatment pre‐ or post‐AMI with corresponding sham and vehicle controls. Temporal changes in left ventricular heme regulatory enzyme expression were measured in response to hemin and AMI. This is the first study to holistically characterize the effects of either AMI and/or hemin on endogenous heme regulation in the myocardium.Hemin markedly increased HMOX1 levels and instigated isoform switching between heme‐synthesizing enzymes in the left ventricle of healthy mice. This isoform switching was further observed with AMI and suggests a progressive increase in heme bioavailability with ischemic injury. Interestingly, HMOX1 — a heme and stress‐inducible enzyme — was induced at only 3 days post‐AMI and did not increase concomitantly with heme‐synthesizing enzymes.Echocardiographic analyses showed preserved left ventricular morphology and cardiac output with hemin administration both pre‐ and post‐AMI. Invasive hemodynamics revealed preserved left ventricular function in mice administered daily hemin when initiated 3h pre‐AMI but not when initiated 2h post‐AMI. Similarly, hemin administration significantly increased H9C2 cardiomyotubule viability when administered prior to H2O2‐mediated injury but not with simultaneous or 2h post‐injury hemin administration. This suggests that time‐dependent mechanisms are involved in conferring hemin‐mediated cardioprotection and could suggest differential mechanisms related to myocyte survival and cardiac remodelling.Support or Funding InformationNatural Sciences and Engineering Research Council of Canada, Nova Scotia Health Research Foundation

Salvia miltiorrhiza and Carthamus tinctorius Extract Prevents Cardiac Fibrosis and Dysfunction after Myocardial Infarction by Epigenetically Inhibiting Smad3 Expression.

The incidence of cardiac dysfunction after myocardial infarction (MI) continues to increase despite advances in treatment. Excessive myocardial fibrosis plays a vital role in the development of adverse cardiac remodeling and deterioration of cardiac function. Understanding the molecular and cellular mechanism of the fibrosis process and developing effective therapeutics are of great importance. Salvia miltiorrhiza and Carthamus tinctorius extract (SCE) is indicated for angina pectoris and other ischemic cardiovascular diseases in China. SCE has been shown to inhibit the platelet activation and aggregation, ameliorate ROS-induced myocardial necrosis by inhibiting mitochondrial permeability transition pore opening, and promote angiogenesis by upregulating the expression of vascular endothelial growth factor (VEGF). However, whether SCE has effect on cardiac fibrosis after MI is not fully clear. Here, a mouse model of MI was established to observe the effect of SCE upon survival, cardiac function, myocardial fibrosis, and inflammation. Quantitative PCR and western blot assays were used to determine the expression of genes related to transforming growth factor-β (TGF-β) cascade and inflammatory responses in vivo. Additionally, the effects of SCE upon the collagen production, TGF-β/Smad3 (SMAD family member 3) signaling, and the levels of histone methylation in primary cardiac fibroblasts were detected. We found that SCE treatment significantly improved survival and left ventricular function in mice after MI. Inhibition of inflammation and fibrosis, as well as decreased expression of Smad3, was observed with SCE treatment. In TGF-β-stimulated cardiac fibroblasts, SCE significantly decreased the expression of collagen, α-smooth muscle actin (α-SMA), and Smad3. Furthermore, SCE treatment downregulated the levels of H3K4 trimethylation (H3K4me3) and H3K36 trimethylation (H3K36me3) at the Smad3 promoter region of cardiac fibroblasts, leading to inhibition of Smad3 transcription. Our findings suggested that SCE prevents myocardial fibrosis and adverse remodeling after MI with a novel mechanism of suppressing histone methylation of the Smad3 promoter and its transcription.

Urocortin 3 Gene Transfer Increases Function of the Failing Murine Heart.

Peptide infusions of peptides the corticotropin releasing factor family, including urocortin 2, stresscopin, and urocortin 3 (UCn3), have favorable acute effects in clinical heart failure (HF), but their short half-lives make them unsuitable for chronic therapy. This study asked whether UCn3 gene transfer, which provides sustained elevation of plasma UCn3 levels, increases the function of the failing heart. HF was induced by transmural left ventricular (LV) cryoinjury in mice. LV function was assessed 3 weeks later by echocardiography. Those with ejection fractions (EF) <40% received intravenous saline or intravenous adeno-associated virus type-8 encoding murine UCn3 (AAV8.mUCn3; 1.9 × 1013 genome copies/kg). Five weeks after randomization, repeat echocardiography, assessment of LV function (+dP/dt, -dP/dt), and quantification of Ca2+ transients and sarcomere shortening in isolated cardiac myocytes were conducted, and assessment of LV Ca2+ handling and stress proteins was performed. Three weeks after myocardial infarction, prior to treatment, EFs were reduced (mean 31%, from 63% in sham-operated animals). Mice randomized to receive UCn3 gene transfer showed increased plasma UCn3 (from 0.1 ± 0.01 ng/mL in the saline group to 5.6 ± 1.1 ng/mL; n = 12 each group; p < 0.0001). Compared to mice that received saline, UCn3 gene transfer was associated with higher values for EF (p = 0.0006); LV +dP/dt (p < 0.0001), and LV -dP/dt (p < 0.0001). Cardiac myocytes from mice that received UCn3 gene transfer showed higher peak Ca2+ transients (p = 0.0005), lower time constant of cytosolic Ca2+ decline (tau, p < 0.0001), and higher rates of sarcomere shortening (+dL/dt, p = 0.03) and lengthening (-dL/dt, p = 0.04). LV samples from mice that received UCn3 gene transfer contained higher levels of SERCA2a (p = 0.0004 vs. HF) and increased amounts of phosphorylated troponin I (p = 0.04 vs. HF). UCn3 gene transfer is associated with improved Ca2+ handling and LV function in mice with HF and reduced EF.

Targeted Gene Therapy with RXFP1 Attenuates Myocardial Infarction and Preserves Left Ventricular Function in Mice

BackgroundSeveral studies have shown that recombinant relaxin targets various mechanisms of cell injury and is protective in models of ischemia/reperfusion (I/R) by reducing oxidative stress, calcium overload, neutrophil activity, NLRP3 inflammasome, endothelial adhesive protein expression, mast cell degranulation, apoptosis, platelet adhesion, as well as improving coronary blood flow. However, there is no information on the role of relaxin receptor 1 (RXFP1) signaling in the heart in the context of myocardial infarction (MI). We sought to determine the effect of RXFP1 gain‐of‐function using novel cardiotropic AAV9 vectors as a therapeutic modality against MI.MethodsRXFP1 cDNA (pCMV6 backbone) was purchased from Origene technologies and introduced into AAV9 cis plasmids following KpnI and AgeI digestion. Plasmids were transformed into NEB stable E. Coli and following propagation, they were transfected into 293T cells with the helper plasmid pDG‐9 to produce AAV9‐RXFP1. Viral particles were isolated post cell lysis and purified. 1.0 × 1011 viral genomes in 100μL were intravenously administered to 8‐week‐old CD1 male mice through tail vein. Four weeks later, a subset of mice was sacrificed and cardiac tissue was extracted for qPCR, western blot, or adult cardiomyocyte isolation. Another subset of mice was subjected to LAD ligation for 30 minutes followed by 24 hours of reperfusion. Cardiac function was assessed via echocardiography prior to sacrifice and infarct size was subsequently determined using triphenyl tetrazolium (TTC) staining.ResultsCardiac tissue from mice injected with AAV9‐RXFP1 showed increased mRNA and protein levels of RXFP1 (Fig. A&amp;B). qPCR analysis also revealed overexpression of RXFP1 in adult cardiomyocytes isolated from mice that received AAV9‐RXFP1 (Fig. C). Phosphorylation of P44/42 MAP kinase downstream of RXFP1 activation was also evident in hearts overexpressing RXFP1 (Fig. D). Moreover, gene therapy with RXFP1 reduced infarct size and preserved cardiac function following MI as compared to control (Fig. E&amp;F).ConclusionOur novel AAV9 construct successfully upregulates cardiac RXFP1, which in turn attenuates infarct size and preserves cardiac function. These findings provide strong rationale for investigating the therapeutic potential of gene therapy with RXFP1 in a more clinically relevant model of established ischemic HF.Support or Funding InformationNHLBI (R01HL133167) and NIA (R21AG053654) to Dr. Salloum.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Inhibition of the cardiac myocyte mineralocorticoid receptor ameliorates doxorubicin-induced cardiotoxicity.

Anthracyclines such as doxorubicin are widely used in cancer therapy but their use is limited by cardiotoxicity. Up to date there is no established strategy for the prevention of anthracyclin-induced heart failure. In this study, we evaluated the role of the cardiac myocyte mineralocorticoid receptor (MR) during doxorubicin-induced cardiotoxicity. A single high-dose or repetitive low-dose doxorubicin administration lead to markedly reduced left ventricular function in mice. Treatment with the MR antagonist eplerenone prevented doxorubicin-induced left ventricular dysfunction. In order to identify the cell types and molecular mechanisms involved in this beneficial effect we used a mouse model with cell type-specific MR deletion in cardiac myocytes. Cardiac myocyte MR deletion largely reproduced the effect of pharmacological MR inhibition on doxorubicin-induced cardiotoxicity. RNAseq from isolated cardiac myocytes revealed a repressive effect of doxorubicin on gene expression which was prevented by MR deletion. We show here that (i) eplerenone prevents doxorubicin-induced left ventricular dysfunction in mice, and (ii) this beneficial effect is related to inhibition of MR in cardiac myocytes. Together with present clinical trial data our findings suggest that MR antagonism may be appropriate for the prevention of doxorubicin-induced cardiotoxicity.

Nanosecond pulsed platelet-rich plasma (nsPRP) improves mechanical and electrical cardiac function following myocardial reperfusion injury.

Ischemia and reperfusion (I/R) of the heart is associated with biochemical and ionic changes that result in cardiac contractile and electrical dysfunction. In rabbits, platelet‐rich plasma activated using nanosecond pulsed electric fields (nsPRP) has been shown to improve left ventricular pumping. Here, we demonstrate that nsPRP causes a similar improvement in mouse left ventricular function. We also show that nsPRP injection recovers electrical activity even before reperfusion begins. To uncover the mechanism of nsPRP action, we studied whether the enhanced left ventricular function in nsPRP rabbit and mouse hearts was associated with increased expression of heat‐shock proteins and altered mitochondrial function under conditions of oxidative stress. Mouse hearts underwent 30 min of global ischemia and 1 h of reperfusion in situ. Rabbit hearts underwent 30 min of ischemia in vivo and were reperfused for 14 days. Hearts treated with nsPRP expressed significantly higher levels of Hsp27 and Hsp70 compared to hearts treated with vehicle. Also, pretreatment of cultured H9c2 cells with nsPRP significantly enhanced the “spare respiratory capacity (SRC)” also referred to as “respiratory reserve capacity” and ATP production in response to the uncoupler FCCP. These results suggest a cardioprotective effect of nsPRP on the ischemic heart during reperfusion.

Myocardial Hypertrophic Remodeling and Impaired Left Ventricular Function in Mice with a Cardiac-Specific Deletion of Janus Kinase 2

The Janus kinase (JAK) system is involved in numerous cell signaling processes and is highly expressed in cardiac tissue. The JAK isoform JAK2 is activated by numerous factors known to influence cardiac function and pathologic conditions. However, although abundant, the role of JAK2 in the regulation or maintenance of cardiac homeostasis remains poorly understood. Using the Cre-loxP system, we generated a cardiac-specific deletion of Jak2 in the mouse to assess the effect on cardiac function with animals followed up for a 4-month period after birth. These animals had marked mortality during this period, although at 4 months mortality in male mice (47%) was substantially higher compared with female mice (30%). Both male and female cardiac Jak2-deleted mice had hypertrophy, dilated cardiomyopathy, and severe left ventricular dysfunction, including a marked reduction in ejection fractions as assessed by serial echocardiography, although the responses in females were somewhat less severe. Defective cardiac function was associated with altered protein levels of sarcoplasmic reticulum calcium-regulatory proteins particularly in hearts from male mice that had depressed levels of SERCA2 and phosphorylated phospholamban. In contrast, SERCA2 was unchanged in hearts of female mice, whereas phosphorylated phospholamban was increased. Our findings suggest that cardiac JAK2 is critical for maintaining normal heart function, and its ablation produces a severe pathologic phenotype composed of myocardial remodeling, heart failure, and pronounced mortality.

The role of 1.5 tesla MRI and anesthetic regimen concerning cardiac analysis in mice with cardiomyopathy.

Accurate assessment of left ventricular function in rodent models is essential for the evaluation of new therapeutic approaches for cardiac diseases. In our study, we provide new insights regarding the role of a 1.5 Tesla (T) magnetic resonance imaging (MRI) device and different anesthetic regimens on data validity. As dedicated small animal MRI and echocardiographic devices are not broadly available, we evaluated whether monitoring cardiac function in small rodents with a clinical 1.5 T MRI device is feasible. On a clinical electrocardiogram (ECG) synchronized 1.5 T MRI scanner we therefore studied cardiac function parameters of mice with chronic virus-induced cardiomyopathy. Thus, reduced left ventricular ejection fraction (LVEF) could be verified compared to healthy controls. However, our results showed a high variability. First, anesthesia with medetomidine, midazolam and fentanyl (MMF) led to depressed cardiac function parameters and more variability than isoflurane gas inhalation anesthesia, especially at high concentrations. Furthermore, calculation of an average ejection fraction value from sequenced scans significantly reduced the variance of the results. To sum up, we introduce the clinical 1.5 T MRI device as a new tool for effective analysis of left ventricular function in mice with cardiomyopathy. Besides, we suggest isoflurane gas inhalation anesthesia at high concentrations for variance reduction and recommend calculation of an average ejection fraction value from multiple sequenced MRI scans to provide valid data and a solid basis for further clinical testing.

68Ga]-Albumin-PET in the Monitoring of Left Ventricular Function in Murine Models of Ischemic and Dilated Cardiomyopathy: Comparison with Cardiac MRI

The purpose of this study is to evaluate left ventricular functional parameters in healthy mice and in different murine models of cardiomyopathy with the novel blood pool (BP) positron emission tomography (PET) tracer [68Ga]-albumin. ECG-gated microPET examinations were obtained in healthy mice, and mice with dilative (DCM) and ischemic cardiomyopathy (ICM) using the novel BP tracer [68Ga]-albumin (AlbBP), as well as [18F]-FDG microPET. Cine-magnetic resonance imaging (MRI) examination performed on a clinical 1.5-T MRI provided the reference standard measurements. When considering the combined group of healthy controls, DCM and ICM AlbBP-PET significantly overestimated the magnitudes of EDV (AlbBP, 181±86 μl; cine-MRI, 125±80 μl; P<0.001) and ESV (AlbBP, 136±92 μl; cine-MRI, 96±77 μl; P<0.001), whereas the EF (AlbBP, 31±16%; cine-MRI, 33±21%; P=0.910) matched closely to cine-MRI results, as did findings with [18F]-FDG. High correlations were found between the measured cardiac parameters (EDV: R=0.978, ESV: R=0.989, and LVEF: R=0.992). Measuring left ventricular function in mice with [68Ga]-albumin BP PET is feasible and showed a high correlation compared to cine-MRI, which was used as a reference standard.

Advancements in pressure–volume catheter technology – stress remodelling after infarction

Microconductance catheters have been successfully applied to measure left ventricular (LV) function in the mouse to assess cardiac or pharmacological interventions for a number of years. New complex admittance methods produce an estimate of the parallel admittance of cardiac muscle that can be used to correct the measurement in real time. This contrasts with existing conductance technologies that require in vivo calibration using a bolus of hypertonic saline. Here, we report the application of this emerging technology in the context of myocardial infarction and LV remodelling. Using a combination of high-resolution ultrasound and LV conductance catheters, we compared measures of LV function using an admittance system and a traditional conductance-derived pressure-volume (PV) system. We subjected C57BL/6 mice to focal myocardial ischaemia-reperfusion by transient ligation of the left anterior descending coronary artery and assessed cardiac function with different systems to determine the reliability and accuracy of these methods to distinguish between normal and dysfunctional ventricle. We demonstrate that the admittance PV system, in our hands, provides a straightforward solution for assessing LV function in mice. Using this technique in combination with other established methods, we measured LV dysfunction following coronary artery occlusion and reperfusion, which can be ameliorated using a known preconditioning agent (CORM-3), and found that functional read-outs are representative of other methods. We have found that, especially in diseased tissue, LV pressure-volume loops derived from complex admittance provide a reproducible and reliable method of determining LV function without the need for technically challenging calibration. Our data suggest that admittance records accurate/physiological LV cavity volumes when compared with other invasive methods in the same animal. This emerging technology is both effective and reproducible for measuring LV function and dysfunction in the mouse, without the need for complicated interventions to calibrate the measurements or training in a new technology. This may mark the way towards a fast and accurate assessment of murine cardiac function in normal animals and disease models.

Analyzing left ventricular function in mice with Doppler echocardiography

Mice are widely used in heart failure research. Accurate evaluation of cardiac structure and function is key to modern cardiovascular research. Doppler echocardiography is a simple, reproducible, and non-invasive method, which allows a longitudinal study of these small animals. Besides common parameters such as left ventricular chamber size, mass, and function, new emerging echo tools are of great interest for small animal imaging. In this review, we describe the technical issues linked to murine cardiovascular anatomy and physiology and the most current echo parameters that can be used.

Global left ventricular function in mice during the development of heart failure

In response to pressure overload the left ventricle (LV) remodels to compensate for the increased workload. Initially this adaptation is beneficial to maintain pump function, but eventually the heart may loose its battle to cope with the increased workload resulting in heart failure (HF). In this study the evolution of global LV function was characterized in a relevant mouse model of LV pressure overload during the development of HF using MRI.

A comparison of retrospectively self-gated magnetic resonance imaging and high-frequency echocardiography for characterization of left ventricular function in mice

Non-invasive imaging methods like echocardiography and magnetic resonance imaging (MRI) are very valuable in longitudinal follow-up studies of cardiac function in small animals. To be able to compare results from studies using different methods, and explain possible differences, it is important to know the agreement between these methods. As both self-gated high-field MRI and high-frequency echocardiography (hf-echo) M-mode are potential methods for evaluation of left ventricular (LV) function in healthy mice, our aim was to assess the agreement between these two methods. Fifteen healthy female C57BL/6J mice underwent both self-gated MRI and hf-echo during the same session of light isoflurane anaesthesia. LV dimensions were estimated offline, and agreement between the methods and reproducibility for the two methods assessed using Bland-Altman methods. In summary, hf-echo M-mode had better inter-observer repeatability than self-gated MRI for all measured parameters. Compared with hf-echo, systolic posterior wall thicknesses were significantly higher when measured by MRI, while diastolic anterior wall thicknesses were found to be significantly smaller. MRI measurements of diastolic LV diameter were also higher using MRI, resulting in larger fractional shortening values compared with the values obtained by hf-echo. In conclusion, hf-echo M-mode is easy to apply, has high temporal and spatial resolution, and good reproducibility. Self-gated MRI might be advantageous in cases of abnormal LV geometry and heterogeneous regional myocardial function, especially with improvements in spatial resolution. The moderate agreement between the methods must be taken into account when comparing studies using the two modalities.

Accelerating global left-ventricular function assessment in mice using reduced slice acquisition and three-dimensional guide-point modelling.

BackgroundTo investigate the utility of three-dimensional guide-point modeling (GPM) to reduce the time required for CMR evaluation of global cardiac function in mice, by reducing the number of image slices required for accurate quantification of left-ventricular (LV) mass and volumes.MethodsFive female C57Bl/6 mice 8 weeks post myocardial infarction induced by permanent occlusion of the left coronary artery, and six male control (un-operated) C57Bl/6 mice, were subject to CMR examination under isoflurane anaesthesia. Contiguous short axis (SAX) slices (1 mm thick 7-9 slices) were obtained together with two long axis (LAX) slices in two chamber and four chamber orientations. Using a mathematical model of the heart to interpolate information between the available slices, GPM LV mass and volumes were determined using full slice (all SAX and two LAX), six slice (four SAX and two LAX) and four slice (two SAX and two LAX) analysis protocols. All results were compared with standard manual volumetric analysis using all SAX slices.ResultsInfarct size was 39.1 ± 5.1% of LV myocardium. No significant differences were found in left ventricular mass and volumes between the standard and GPM full and six slice protocols in infarcted mice (113 ± 10, 116 ± 11, and 117 ± 11 mg respectively for mass), or between the standard and GPM full, six and four slice protocols in control mice, (105 ± 14, 106 ± 10, 104 ± 12, and 105 ± 7 mg respectively for mass). Significant differences were found in LV mass (135 ± 18 mg) and EF using the GPM four slice protocol in infarcted mice (p < 0.05).ConclusionGPM enables accurate analysis of LV function in mice with relatively large infarcts using a reduced six slice acquisition protocol, and in mice with normal/symmetrical left-ventricular topology using a four slice protocol.

The effects of VEGF-R1 and VEGF-R2 ligands on angiogenic responses and left ventricular function in mice

Vascular endothelial growth factors (VEGFs) and their receptors (VEGF-Rs) are among the most powerful factors regulating vascular growth. However, it has remained unknown whether stimulation of VEGF-R1, VEGF-R2 or both of the receptors produces the best angiogenic responses in myocardium. The aim of this study was to compare the VEGF-R1-specific ligand VEGF-B(186), VEGF-R2-specific ligand VEGF-E and VEGF-A(165,) which stimulates both receptors, regarding their effects on angiogenesis and left ventricular function in mice. High-resolution echocardiography was used to guide the closed-chest injections of adenoviral (Ad) vectors expressing VEGF-B(186,) VEGF-E, and VEGF-A(165) into the anterior wall of the left ventricle in C57Bl/6J mice. Angiogenic and functional effects were analysed using histology, ultrasound and perfusion analyses 6 (D6) and 14 (D14) days after the Ad injection. AdVEGF-A(165) induced a strong angiogenic response seen as an enlargement of myocardial capillaries whereas angiogenesis induced by AdVEGF-B(186) and AdVEGF-E seemed more physiological. The increase in the capillary area was accompanied with an increase in myocardial perfusion at D6 after the gene injection. AdVEGF-A(165) and AdVEGF-E induced endothelial-specific proliferation whereas AdVEGF-B(186) mostly induced proliferation of cardiomyocytes. AdVEGF-A(165) induced more pronounced tissue damage than AdVEGF-B(186) and AdVEGF-E. Left ventricular function measured as ejection fraction did not change during the follow-up. AdVEGF-A(165) increased both VEGF-R1 and VEGF-R2 protein expression whereas AdVEGF-B(186) and AdVEGF-E did not affect endogenous receptor expression levels. AdVEGF-B(186) and AdVEGF-E are equally potent in inducing therapeutic angiogenesis in mouse myocardium and produce less side effects than AdVEGF-A(165).

Comparative Study of High-resolution Microimaging with 30-MHz Scanner for Evaluating Cardiac Function in Mice

The accurate assessment of cardiac function in mice is challenging because of their small heart size and rapid heart rate. We examined the usefulness of novel high-resolution echocardiography (HRE) with a 30-MHz transducer in evaluating cardiac function in 20 mice compared with conventional echocardiography (CE) with a 13-MHz transducer. The left ventricular (LV) regional wall motion (RWM), LV end-diastolic dimension, fractional shortening, anterior LV wall thickness, E/A, and myocardial performance index were assessed. RWM analysis was more feasible by HRE than by CE (P < .05). Interobserver agreement in RWM analysis and correlation in LV end-diastolic dimension, fractional shortening, anterior LV wall thickness, E/A, and myocardial performance index were all better with HRE than CE. HRE is superior to CE in assessing LV function in mice. HRE is potentially a useful method for accurate assessment of cardiac function in various mice models.

Validation of the wall motion score and myocardial performance indexes as novel techniques to assess cardiac function in mice after myocardial infarction

The aim of this study was to determine the feasibility and accuracy of wall motion score index (WMSI) and myocardial performance index (MPI) for measuring regional and global left ventricular (LV) function with use of high-resolution echocardiography after myocardial infarction (MI) in mice. In 48 mice, myocardial infarction was induced by ligation in the middle of the left anterior descending coronary artery. Echocardiography was performed under anesthesia at baseline and 1 mo after MI. WMSI was analyzed by a 16-segment model on short-axis views, and wall motion was scored as 1 for normal, 2 for hypokinetic, 3 for akinetic, 4 for dyskinetic, and 5 for aneurysmal. WMSI was calculated as the sum of scores divided by the total number of segments. MPI was calculated on the basis of isovolumetric contraction time (IVCT), isovolumetric relaxation time (IVRT), and ejection time (ET): MPI = (IVCT + IVRT)/ET. We measured LV ejection fraction (LVEF), end-systolic and end-diastolic volumes (ESV and EDV), fractional shortening (FS), and infarct size (IS). LVEF at 4 wk after MI was reduced at 32.8 +/- 9.0%. Linear correlation analyses showed that WMSI (1.6 +/- 0.3) correlated with LVEF (r = -0.84, P < 0.0005), FS (r = -0.43, P = 0.003), and IS (34.3 +/- 15.3%, r = 0.86, P < 0.0005). MPI (0.67 +/- 0.09) correlated with LVEF (r = -0.67, P < 0.0005) and IS (r = 0.72, P < 0.0005). MPI also correlated with mitral inflow velocity (r = -0.68, P < 0.0005) and deceleration time (r = -0.42, P = 0.003). Stepwise regression analysis revealed that WMSI was independently associated with IS. IS, FS, mitral inflow velocity, and deceleration time were independent determinants of MPI. In conclusion, echocardiographic assessments of WMSI and MPI in mice are feasible and correlate strongly with two-dimensional measurement of LV function and IS. These novel parameters provide additional noninvasive assessment of regional and global LV function in mice after MI.

Commentary on „Direct comparison of magnetic resonance imaging and conductance microcatheter in the evaluation of left ventricular function in mice“ by Jacoby et al.

Commentary on „Direct comparison of magnetic resonance imaging and conductance microcatheter in the evaluation of left ventricular function in mice“ by Jacoby et al.