Method Article

Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography

DOI:

10.3791/51041

February 3rd, 2014

In This Article

Summary

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Right ventricle (RV) dysfunction is critical to the pathogenesis of cardiovascular disease, yet limited methodologies are available for its evaluation. Recent advances in ultrasound imaging provide a noninvasive and accurate option for longitudinal RV study. Herein, we detail a step-by-step echocardiographic method using a murine model of RV pressure overload.

Abstract

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Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, the RV is significantly affected in pulmonary diseases such as pulmonary artery hypertension (PAH). In addition, the RV is remarkably sensitive to cardiac pathologies, including left ventricular (LV) dysfunction, valvular disease or RV infarction4. To understand the role of RV in the pathogenesis of cardiac diseases, a reliable and noninvasive method to access the RV structurally and functionally is essential.

A noninvasive trans-thoracic echocardiography (TTE) based methodology was established and validated for monitoring dynamic changes in RV structure and function in adult mice. To impose RV stress, we employed a surgical model of pulmonary artery constriction (PAC) and measured the RV response over a 7-day period using a high-frequency ultrasound microimaging system. Sham operated mice were used as controls. Images were acquired in lightly anesthetized mice at baseline (before surgery), day 0 (immediately post-surgery), day 3, and day 7 (post-surgery). Data was analyzed offline using software.

Several acoustic windows (B, M, and Color Doppler modes), which can be consistently obtained in mice, allowed for reliable and reproducible measurement of RV structure (including RV wall thickness, end-diastolic and end-systolic dimensions), and function (fractional area change, fractional shortening, PA peak velocity, and peak pressure gradient) in normal mice and following PAC.

Using this method, the pressure-gradient resulting from PAC was accurately measured in real-time using Color Doppler mode and was comparable to direct pressure measurements performed with a Millar high-fidelity microtip catheter. Taken together, these data demonstrate that RV measurements obtained from various complimentary views using echocardiography are reliable, reproducible and can provide insights regarding RV structure and function. This method will enable a better understanding of the role of RV cardiac dysfunction.

Introduction

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Historically, prognostic assessment of heart failure has focused on the LV, which is easy to image via echocardiography. Numerous studies on LV structure and function using echocardiography have led to the establishment of normal values for LV structure and function1,5,6. Measurements of LV size and systolic function obtained from two-dimensional and Color Doppler images are of great importance as they allow visual delineation of compartments and geometry in great detail for the LV7. M-Mode is often used for measuring LV dimensions and fractional shortening (FS) in mice. Inter-observer and intra-observer variability are low for diameter measureme....

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Protocol

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Surgical Procedure

  1. Obtain 8 week-old male C57BL/6 mice and acclimate for one week before any experimental procedures are performed.
  2. Prior to imaging, pulmonary artery occlusion is performed as described previously14 in accordance with AVMA guidelines and approved IACUC protocols.
    Echocardiographic Images Acquisition and Measurements
    All abbreviations used are summarized in Table 1.

1. Parasternal Long Axis (PLAX) M Mode View to Obtain RV Chamber Dimension, Fractional Shortening (FS), and RV Wall Thickness

  1. Use B Mode setting to obtain a full LV par....

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Results

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In this study, baseline echocardiography was performed 48 hr prior to surgery. Mice were randomized into two groups. Mice received pulmonary artery occlusions (PAC) and sham operations (Sham). Echocardiography was performed at day 0, 3, and 7 following surgical procedure.  The animals were euthanized immediately following the last echocardiography and hearts were harvested for histological assessment. Catheterization was conducted in subgroup (n=3 and 2 for day 0 and 7, respectively) of PAC mice to measure RVSP via press.......

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Discussion

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We demonstrate that TTE provides a sensitive and reproducible methodology for routine assessment of RV structure and function in mice. Before the advent of TTE, studies of the RV largely focused on RVSP measurement via right heart catheterization, a terminal and invasive procedure6,9,11,17.

Prior reports have described a variety of techniques for performing right heart measurements3,4,11,17-19. However, the majority of previous studies reported RV size and structural data.......

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Disclosures

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There is nothing to disclose.

Acknowledgements

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We thank Fred Roberts and Chris White for exemplary technical support. We thank Brigham Women’s Hospital Cardiovascular Physiology Core for providing with the instrumentation and the funds for this work. This work was supported in part by NHLBI grants HL093148, HL086967, and HL 088533(RL), K99HL107642 and the Ellison Foundation (SC).

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
High Frequency UltrasoundFUJIFILM VisualSonics, Inc.Vevo 2100
High-frequency Mechanical TransducerFUJIFILM VisualSonics, Inc.MS250, MS550D, MS400
Millar Mikro Pressure CatheterMillarSPR-1000

References

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  1. Anavekar, N. S., et al. Usefulness of right ventricular fractional area change to predict death, heart failure, and stroke following myocardial infarction (from the VALIANT ECHO Study). Am. J. Cardiol. 101, 607-612 (2008).
  2. Berger, R. M., Cromme-Dijkhuis, A. H., Witsenburg, M., Hess, J.

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Tags

Right Ventricular FunctionPulmonary Artery ConstrictionTransthoracic EchocardiographyMouse ModelRV Wall ThicknessFractional Area ChangeColor Doppler ModeHigh Frequency UltrasoundParasternal Long AxisPulmonary Artery Peak Velocity

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