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Establishment and Confirmation of a Postnatal Right Ventricular Volume Overload Mouse Model

Published: June 9th, 2023



* These authors contributed equally

This protocol presents the establishment and confirmation of a postnatal right ventricular volume overload (VO) model in mice with abdominal arteriovenous fistula (AVF), which can be applied to investigate how VO contributes to postnatal heart development.

Right ventricular (RV) volume overload (VO) is common in children with congenital heart disease. In view of distinct developmental stages,the RV myocardium may respond differently to VO in children compared to adults. The present study aims to establish a postnatal RV VO model in mice using a modified abdominal arteriovenous fistula. To confirm the creation of VO and the following morphological and hemodynamic changes of the RV, abdominal ultrasound, echocardiography, and histochemical staining were performed for 3 months. As a result, the procedure in postnatal mice showed an acceptable survival and fistula success rate. In VO mice, the RV cavity was enlarged with a thickened free wall, and the stroke volume was increased by about 30%-40% within 2 months after surgery. Thereafter, the RV systolic pressure increased, corresponding pulmonary valve regurgitation was observed, and small pulmonary artery remodeling appeared. In conclusion, modified arteriovenous fistula (AVF) surgery is feasible to establish the RV VO model in postnatal mice. Considering the probability of fistula closure and elevated pulmonary artery resistance, abdominal ultrasound and echocardiography must be performed to confirm the model status before application.

Right ventricular (RV) volume overload (VO) is common in children with congenital heart disease (CHD), which leads to pathological myocardial remodeling and a poor long-term prognosis1,2,3. An in-depth understanding of RV remodeling and related early targeted interventions is essential for a good outcome in children with CHD. There are several differences in the molecular structures, physiological functions, and responses to stimuli in the hearts of adults and children1,4,5,

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All of the procedures presented here conformed to the principles outlined in the Declaration of Helsinki and were approved by the Animal Welfare and Human Studies Committee at Shanghai Children's Medical Center (SCMC-LAWEC-2023-003). C57BL/6 mice pups (P7, males, 3-4 g) were used for the present study. The animals were obtained from a commercial source (see Table of Materials). The mice pups and their nursing mothers (pups:mothers = 6:1 in a single cage) were kept under specific-pathogen-free laborat.......

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Survival rate and AVF patency within 3 months
A total of 30 (75%) mice in the VO group and 19 (95%) mice in the sham group survived the AVF surgery (Figure 4A). In the VO group, eight mice died within 1 day after surgery due to excessive bleeding (n = 5) or cannibalization (n = 3), whereas two mice died of unknown causes at 1 month.

Of the surviving VO mice (n = 30), ultrasound confirmed the successful establishment of fistulas in 21 mice po.......

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Previously, the classic RV VO model was created using valve regurgitation21; however, compared to AVF, open-heart valve surgery may require more sophisticated techniques and may be associated with significantly higher mortality, particularly in postnatal mice. As animal studies have shown that the same effect of VO has been achieved by AVF22, modified abdominal fistula surgery with less trauma was used in this study.

Certain factors were consider.......

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This work was supported by the National Science Foundation of China (no. 82200309) and the Innovation Project of Distinguished Medical Team in Ningbo (no. 2022020405)


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NameCompanyCatalog NumberComments
70% EthanolTiandz,Chia
ACETAMINOPHEN Oral SolutionVistaPharm, Inc. Largo, FL 33771, USANDC 66689-054-01
Anesthesia machineRWD Life Science,ChinaR550IP
Anesthesia maskRWD Life Science,China68680
C57BL/6 miceXipu’er-bikai Experimental Animal Co., Ltd (Shanghai, China)
Hair removal creamVeet, FranceVT-200
Hematoxylin and eosin Kit Beyotime biotech C0105M 
IsofluraneRWD Life Science,ChinaR510-22-10
Microscope Yuyan Instruments, ChinaSM-301
Surgical suture needlesNINGBO MEDICAL NEEDLE CO.,LTD, China
Thermostatic heating platformQingdao Juchuang Environmental Protection Group Co., Ltd, China
Ultrasound deviceFUJIFILM VisualSonics, Inc.Vevo 2100Image modes includes B-Mode, Color Doppler Mode and Pulsed Wave Doppler Mode
Ultrasound gelParker Laboratories,United StatesREF 01-08
Ultrasound transducerFUJIFILM VisualSonics, Inc.MS 400

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