Name: induction of right ventricular failure by pulmonary artery constriction and evaluation of right ventricular function in mice
Uploaded: 2019-05-13T13:30:00
Description: Watch this Scientific Journal Video about Induction of Right Ventricular Failure by Pulmonary Artery Constriction and Evaluation of Right Ventricular Function in Mice at JoVE.com

This work was supported by grants from the National Natural Science Foundation of China (81570464, 81770271; to Dr. Liao) and the Municipal Planning Projects of Scientific Technology of Guangzhou (201804020083) (to Dr. Liao).

All procedures were performed in accordance with institutional guidelines for animal research, which conform to the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised in 1996). C57BL/6 male mice (8-10 weeks old, weighing 20-25 g) were provided by the Animal Center of South Medical University. After arrival, the mice were housed under a 12/12 h dark/light cycle, with sufficient food and water.
1. Preparation of th...

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Pathological increases in RV filling pressures result in a leftward shift of the septum, which can alter the LV geometry21. These changes contribute to reduced cardiac output and LV ejection fraction (LVEF), which can cause a hemodynamic disorder of the circulatory system22. Therefore, an efficient, stable, and economical model for studying the mechanism of RVF is valuable.
We developed a more effective and highly reproducible approach to PAC usi...

RV dysfunction (RVD), defined here as evidence of an abnormal RV structure or function, is associated with poor clinical outcomes. RVF, as the end stage of RV function, is a clinical syndrome with signs and symptoms of heart failure that result from progressive RVD1. With differences in structure and physiological function, left ventricular (LV) failure and RVF have different pathophysiological mechanisms. A few independent pathophysiological mechanisms in RVF have been reported, including overexpression of &#946;2-adrenergic receptor signaling2, inflammation3, transverse tubule remodeling, and Ca2+ handling dysfunction4.
RVF can be caused by volume or pressure overload of the RV. Previous animal models have used SU5416 (a potent and selective inhibitor of the vascular endothelial growth factor receptor) combined with hypoxia (SuHx)5,6 or monocrotaline7 to induce pulmonary hypertension, which results in RVF secondary to pulmonary vascular disease2. The researchers conducting these studies focused on the vasculature instead of the pathological progression of RVF. Moreover, monocrotaline has extra-cardiac effects that cannot precisely represent cardiogenic disease. Other models have used arteriovenous shunts to induce volume overload and RVF8. However, this surgery is difficult to perform and inappropriate for mice, who require long induction periods for the production of RVF.
Rat PAC models using banding clips also exist9,10. Compared with rats, mice have many advantages as animal models of cardiac diseases, such as easier reproduction, more widespread use, reduced costs, and access to gene modification11. However, the diameters of the banding clips usually range from 0.5 mm to 1.0 mm, which are too large for mice9. In addition, the banding clip is hard to produce, imitate, and popularize in other labs.
We provide a protocol to develop a modified reproductive RVF mouse model based on reported studies, which uses PAC to mimic the tetralogy of Fallot and Noonan syndrome or other pulmonary arterial hypertensive diseases12,13,14,15,16,17,18,19. This PAC approach is created by ligating the pulmonary trunk of mice using a latch and padding needle made inhouse to control the degree of constriction. The latch needle is made of a 90&#176; curved injection syringe with a braided silk suture passed through the syringe. The needle is made from common materials using a process that is easy to master. The padding needle is curved 120&#176; from the gauge needle. Padding needles with different diameters (0.6-0.8 mm) are used, depending on the mice&#39;s weight (20-35 g). Additionally, we establish an evaluation criterion to determine the stability and quality of the RVF model by echocardiography and right heart catheterization. We use mice as the model animal because of their widespread use in other experiments. The needles made in the lab are easy to reproduce and can be widely used in other labs. This study provides a good approach for researchers to investigate the mechanism of RVF.

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			<td height="21" style="height:21px;width:395px;">ALC-V8S ventilator</td>
			<td style="width:310px;">SHANGHAI&#160; ALCOTT&#160; BIOTECH&#160; CO</td>
			<td style="width:123px;">ALC-V8S</td>
			<td style="width:327px;">Assist ventilation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Animal Mini Ventilator</td>
			<td style="width:310px;">Haverd</td>
			<td style="width:123px;">Type 845</td>
			<td style="width:327px;">Assist ventilation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Animal ultrasound system VEVO2100</td>
			<td style="width:310px;">Visual Sonic</td>
			<td style="width:123px;">&#160;VEVO2100</td>
			<td style="width:327px;">Echocardiography</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Cold light illuminator</td>
			<td style="width:310px;">Olympus</td>
			<td style="width:123px;">ILD-2</td>
			<td style="width:327px;">Light</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Heat pad- thermostatic surgical system (ALC-HTP-S1)</td>
			<td style="width:310px;">SHANGHAI&#160; ALCOTT&#160; BIOTECH&#160; CO</td>
			<td style="width:123px;">ALC-HTP-S1</td>
			<td style="width:327px;">Heating</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Isoflurane</td>
			<td style="width:310px;">RWD life science</td>
			<td style="width:123px;">R510-22</td>
			<td style="width:327px;">Inhalant anaesthesia</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Matrx VIP 3000 Isofurane Vaporizer</td>
			<td style="width:310px;">Midmark Corporation</td>
			<td style="width:123px;">VIP 3000</td>
			<td style="width:327px;">Anesthetization</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:395px;">Medical braided silk suture (6-0)</td>
			<td style="width:310px;">Shanghai Pudong Jinhuan Medical Supplies Co.</td>
			<td style="width:123px;">6-0</td>
			<td style="width:327px;">Ligation</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Medical nylon suture (5-0)</td>
			<td style="width:310px;">Ningbo Medical Needle Co.</td>
			<td style="width:123px;">5-0</td>
			<td style="width:327px;">Suture</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Millar Catheter (1.0 F)</td>
			<td style="width:310px;">AD instruments</td>
			<td style="width:123px;">1.0F</td>
			<td style="width:327px;">For right heart catheterization</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Pentobarbital sodium salt</td>
			<td style="width:310px;">Merck</td>
			<td style="width:123px;">25MG</td>
			<td style="width:327px;">Anesthetization</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:395px;">PowerLab multi-Directional physiological Recording System</td>
			<td style="width:310px;">AD instruments</td>
			<td style="width:123px;">4/35</td>
			<td style="width:327px;">Record the result of right heart catheterization</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Precision electronic balance</td>
			<td style="width:310px;">Denver Instrument</td>
			<td style="width:123px;">TB-114</td>
			<td style="width:327px;">Weighing sensor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Self-made latch needle</td>
			<td style="width:310px;"></td>
			<td style="width:123px;"></td>
			<td style="width:327px;">Separate the aorta and pulmonary trunk</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Self-made padding needle&#160;</td>
			<td style="width:310px;"></td>
			<td style="width:123px;"></td>
			<td style="width:327px;">Constriction</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Self-made tracheal intubation</td>
			<td style="width:310px;"></td>
			<td style="width:123px;"></td>
			<td style="width:327px;">Tracheal intubation&#160;</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Small animal microsurgery equipment</td>
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induction of right ventricular failure by pulmonary artery constriction and evaluation of right ventricular function in mice

The mechanism of right ventricular failure (RVF) requires clarification due to the uniqueness, high morbidity, high mortality, and refractory nature of RVF. Previous rat models imitating RVF progression have been described. Compared with rats, mice are more accessible, economical, and widely used in animal experiments. We developed a pulmonary artery constriction (PAC) approach which is comprised of banding the pulmonary trunk in mice to induce right ventricular (RV) hypertrophy. A special surgical latch needle was designed that allows for easier separation of the aorta and the pulmonary trunk. In our experiments, the use of this fabricated latch needle reduced the risk of arteriorrhexis and improved the surgical success rate to 90%. We used different padding needle diameters to precisely create quantitative constriction, which can induce different degrees of RV hypertrophy. We quantified the degree of constriction by evaluating the blood flow velocity of the PA, which was measured by noninvasive transthoracic echocardiography. RV function was precisely evaluated by right heart catheterization at 8 weeks after surgery. The surgical instruments made inhouse were composed of common materials using a simple process that is easy to master. Therefore, the PAC approach described here is easy to imitate using instruments made in the lab and can be widely used in other labs. This study presents a modified PAC approach that has a higher success rate than other models and an 8-week postsurgery survival rate of 97.8%. This PAC approach provides a useful technique for studying the mechanism of RVF and will enable an increased understanding of RVF.

In this study, mice were randomly assigned to the PAC group (n = 9) or the sham operation group (n = 10). Echocardiography was performed at 1, 4, and 8 weeks after the surgery. Eight weeks after surgery, following the last echocardiography and catheterization assessments, the mice were euthanized, and their hearts were harvested for a morphological and histological assessment.
Pulmonary trunk constriction cause...

Watch this Scientific Journal Video about Induction of Right Ventricular Failure by Pulmonary Artery Constriction and Evaluation of Right Ventricular Function in Mice at JoVE.com

Induction of Right Ventricular Failure by Pulmonary Artery Constriction and Evaluation of Right Ventricular Function in Mice

Here, we provide a useful approach for studying the mechanism of right ventricular failure. A more convenient and efficient approach to pulmonary artery constriction is established using surgical instruments made inhouse. In addition, methods to evaluate the quality of this approach by echocardiography and catheterization are provided.

The mechanism of right ventricular failure (RVF) requires clarification due to the uniqueness, high morbidity, high mortality, and refractory nature of ...

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