Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Aug 17;10(16):e020854.
doi: 10.1161/JAHA.121.020854. Epub 2021 Aug 13.

Postnatal Right Ventricular Developmental Track Changed by Volume Overload

Sijuan Sun  1 , Yuqing Hu  2 , Yingying Xiao  3 , Shoubao Wang  4 , Chuan Jiang  3 , Jinfen Liu  3 , Hao Zhang  3   5 , Haifa Hong  5 , Fen Li  2 , Lincai Ye  3   6   5
Affiliations

Postnatal Right Ventricular Developmental Track Changed by Volume Overload

Sijuan Sun et al. J Am Heart Assoc. .

Abstract

Background Current right ventricular (RV) volume overload (VO) is established in adult mice. There are no neonatal mouse VO models and how VO affects postnatal RV development is largely unknown. Methods and Results Neonatal VO was induced by the fistula between abdominal aorta and inferior vena cava on postnatal day 7 and confirmed by abdominal ultrasound, echocardiography, and hematoxylin and eosin staining. The RNA-sequencing results showed that the top 5 most enriched gene ontology terms in normal RV development were energy derivation by oxidation of organic compounds, generation of precursor metabolites and energy, cellular respiration, striated muscle tissue development, and muscle organ development. Under the influence of VO, the top 5 most enriched gene ontology terms were angiogenesis, regulation of cytoskeleton organization, regulation of vasculature development, regulation of mitotic cell cycle, and regulation of the actin filament-based process. The top 3 enriched signaling pathways for the normal RV development were PPAR signaling pathway, citrate cycle (Tricarboxylic acid cycle), and fatty acid degradation. VO changed the signaling pathways to focal adhesion, the PI3K-Akt signaling pathway, and pathways in cancer. The RNA sequencing results were confirmed by the examination of the markers of metabolic and cardiac muscle maturation and the markers of cell cycle and angiogenesis. Conclusions A neonatal mouse VO model was successfully established, and the main processes of postnatal RV development were metabolic and cardiac muscle maturation, and VO changed that to angiogenesis and cell cycle regulation.

Keywords: RNA sequencing; cardiomyocyte; proliferation; right ventricle; volume overload.

PubMed Disclaimer

Conflict of interest statement

None.

Figures

Figure 1
Figure 1. Establishment of the abdominal aorta and inferior vena cava fistula.
A, Schematic diagram of abdominal aorta and inferior vena cava fistula model. B, Under normal circumstances, the inferior vena cava has no pulsatile blood flow, and the detection point is located in the inferior vena cava. C, Under normal circumstances, there are pulsatile blood flow in abdominal aorta, with a peak blood flow velocity of 400 mm/s. D, The representative image of blood flow through the puncture point. E, The representative image of pulsating blood flow at the puncture point, with a peak blood flow velocity of 450 mm/s, which is higher than that of normal aortic blood flow. F, After puncture, the pulsatile blood flow appeared in the inferior vena cava, with a peak blood flow velocity of 200 mm/s. AA indicates abdominal aorta; DP, detection point; IVC, inferior vena cava; and PP, puncture point.
Figure 2
Figure 2. Volume overload (VO) increased in the abdominal aorta and inferior vena cava fistula model.
A, The representative echo image showed that 2 weeks after puncture, the pulmonary artery velocity and velocity time integral in the VO group increased. B, Quantitative statistics of pulmonary artery‐velocity and velocity time integral in sham and VO group, n=6, Student t‐test. C, Hematoxylin and eosin staining showed that 12 months after of puncture, the free wall of the right ventricle was thickened in the VO group, and the ventricular septum shifted to the left. D, Quantification of the thickness of the free wall of the right ventricle, n=6, Student t‐test. LV indicates left ventricle; PA, pulmonary artery; RV, right ventricle; VO, volume overload; and VTI, velocity time integral.
Figure 3
Figure 3. The transcriptomic changes of postnatal right ventricular development.
A, Volcano map of differentially expressed genes of postnatal right ventricular development in the normal condition (postnatal day 21 sham vs postnatal day 14 sham) and under the influence of volume overload (postnatal day 21 volume overload and postnatal day 14 volume overload). B, Cluster analysis of differentially expressed genes. Every group had 3 mice. The redder the color, the higher the expression level; the bluer, the lower. The clusters of genes in each group are quite different from each other but similar in the same group. C, Venn diagram of differentially expressed genes. P14 indicates postnatal day 14; and P21, postnatal day 21.
Figure 4
Figure 4. The processes of normal postnatal right ventricular development were primarily associated with metabolic and cardiac muscle maturation, as indicated by the gene ontology (GO) analysis.
A, From the results of the GO enrichment analysis, the most significant 10 terms are displayed. The abscissa is the GO term, and the ordinate is the significance level of GO term enrichment. The higher the value, the more significant the result, and the different colors represent 3 different GO subclasses: biological process, cellular component, and molecular function. B, From the results of the GO enrichment analysis, we selected the most significant 30 terms to draw scatter plots for display. The abscissa is the ratio of the number of differentially expressed genes on the GO term to the total number of differentially expressed genes, the ordinate is GO term, the size of the dots represents the number of genes annotated to the GO term, and the color from red to purple represents the significance level of GO term enrichment. BP indicates biological process; CC, cellular component; and MF, molecular function; P14, postnatal day 14; P21, postnatal day 2; and VO, volume overload.
Figure 5
Figure 5. The processes of postnatal right ventricular development partially switched to angiogenesis and cell cycle regulation, as indicated by gene ontology (GO) analysis.
A, From the results of the GO enrichment analysis, the most significant 10 terms are displayed. The abscissa is the GO term, and the ordinate is the significance level of the GO term enrichment. The higher the value, the more significant the result, and the different colors represent 3 different GO subclasses: biological process, cellular component, and molecular function. B, From the results of the GO enrichment analysis, the most significant 30 terms were selected to draw scatter plots for display. The abscissa is the ratio of the number of differentially expressed genes on the GO term to the total number of differentially expressed genes, the ordinate is GO term, the size of the dots represent the number of genes annotated to the GO term, and the colors from red to purple represent the significance level of the GO term enrichment. P14 indicates postnatal day 14; BP, biological process; CC, cellular component; MF, molecular function; and P21, postnatal day 2.
Figure 6
Figure 6. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the differentially expressed genes of normal postnatal right ventricular (RV) development and the postnatal RV development under the influence of volume overload.
A, The KEGG pathways of normal RV development. The most significant 20 KEGG pathways from the KEGG enrichment results are displayed. The abscissa is the KEGG pathway, and the ordinate is the significance level of pathway enrichment. The higher the value, the greater the significance. B, The KEGG pathway of postnatal RV development under the influence of volume overload. The most significant 20 KEGG pathways from the KEGG enrichment results are displayed. The abscissa is the KEGG pathway, and the ordinate is the significance level of pathway enrichment. The higher the value, the greater the significance. C, The KEGG pathways of normal RV development. From the KEGG enrichment results, the most significant 20 KEGG pathways were selected for the scatter plots. The abscissa is the ratio of the number of differentially expressed genes on the KEGG pathway to the total number of differentially expressed genes, the ordinate is the KEGG pathway, the size of the dots represents the number of genes annotated to the KEGG pathway, and the color from red to purple the represents significance level of KEGG pathway enrichment. D, The KEGG pathways of postnatal RV development under the influence of volume overload. From the KEGG enrichment results, the most significant 20 KEGG pathways were selected for the scatter plots. The abscissa is the ratio of the number of differentially expressed genes on the KEGG pathway to the total number of differentially expressed genes, the ordinate is the KEGG pathway, the size of the dots represents the number of genes annotated to the KEGG pathway, and the color from red to purple the represents significance level of KEGG pathway enrichment. P14 indicates postnatal day 14; P21, postnatal day 2; and VO, volume overload.
Figure 7
Figure 7. The processes of normal postnatal right ventricular (RV) development are primarily associated with metabolic and cardiac muscle maturation.
A, The top 20‐fold change genes between postnatal day 21 (P21) sham and postnatal day 14 (P14) sham were selected for verification by quantitative real‐time polymerase chain reaction. P<0.05, n=3 mice, Mann‐Whitney test. B, Representative isolated cardiomyocytes from P14 and P21 RV. Sarcomeric α‐actinin (SAA, white), 4',6‐diamidino‐2‐phenylindole (blue). C, Quantification of cell width from P14 and P21 RV, Student t test. D, Quantification of cell length from P14 and P21 RV. E, Quantification of cell area from P14 and P21 RV. F, Representative of T‐tubule and mitochondria from P14 and P21 RV. MM4‐64 (T‐tubule, white), mitochondria (red). G, Quantification of mitochondria intensity from P14 and P21 RV, Student t‐test. H, Representative of the calcium transient image of cardiomyocytes from P14 and P21 RV, Student t test. I, Plot representative of the calcium transient image of cardiomyocytes from P14 and P21 RV. J, Quantification of the calcium transient parameters (amplitude (Amp) and time to peak), Student t‐test. P14, postnatal day 14; and P21, postnatal day 2.
Figure 8
Figure 8. The processes of postnatal right ventricular development partially switched to cell cycle regulation and angiogenesis.
A, The top 20‐fold change genes between postnatal day 21 volume overload and postnatal day 14 volume overload were selected for verification by quantitative real‐time polymerase chain reaction. P<0.05, n=3 mice, Mann‐Whitney test. B, Representative Ki67‐positive cardiomyocytes. Sarcomeric α‐actinin (red), 4',6‐diamidino‐2‐phenylindole (blue), Ki67 (green). Star indicates Ki67‐positive non‐cardiomyocytes; arrow indicates Ki67‐positive cardiomyocytes. C, Quantification of Ki67‐positive cardiomyocytes, n=6 mice, Student t‐test. D, Representative pHH3 (phospho‐histone H3)‐positive cardiomyocytes from postnatal day 14 volume overload and postnatal day 21 volume overload. Sarcomeric α‐actinin (red), 4',6‐diamidino‐2‐phenylindole (blue), pHH3 (green). E, Quantification of pHH3‐positive cardiomyocytes, n=6 mice, Student t‐test. F, Representative aurora B‐positive cardiomyocytes from postnatal day 14 volume overload and postnatal day 21 volume overload . Sarcomeric α‐actinin (red), diamidino‐2‐phenylindole (blue), aurora B (green). G, Quantification of aurora B‐positive cardiomyocytes, n=60 sections from 6 mice, Student t‐test. H, Representative CD31‐positive cells. I, Quantification of CD31‐positive cells, n=30 sections from 6 mice, Student t‐test. DAPI indicates diamidino‐2‐phenylindole; P14, postnatal day 14; P21, postnatal day 2; SAA, sarcomeric α‐actinin; and VO, volume overload.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Latus H, Nassar MS, Wong J, Hachmann P, Bellsham‐Revell H, Hussain T, Apitz C, Salih C, Austin C, Anderson D, et al. Ventricular function and vascular dimensions after Norwood and hybrid palliation of hypoplastic left heart syndrome. Heart. 2018;104:244–252. DOI: 10.1136/heartjnl-2017-311532. - DOI - PubMed
    1. Borrelli N, Di Salvo G, Sabatino J, Ibrahim A, Avesani M, Sirico D, Josen M, Penco M, Fraisse A, Michielon G, et al. Serial changes in longitudinal strain are associated with outcome in children with hypoplastic left heart syndrome. Int J Cardiol. 2020;317:56–62. DOI: 10.1016/j.ijcard.2020.03.085. - DOI - PubMed
    1. Warnes CA. Transposition of the great arteries. Circulation. 2006;114:2699–2709. DOI: 10.1161/CIRCULATIONAHA.105.592352. - DOI - PubMed
    1. Broberg CS, Valente AM, Huang J, Burchill LJ, Holt J, Van Woerkom R, Powell AJ, Pantely GA, Jerosch‐Herold M. Myocardial fibrosis and its relation to adverse outcome in transposition of the great arteries with a systemic right ventricle. Int J Cardiol. 2018;271:60–65. DOI: 10.1016/j.ijcard.2018.04.089. - DOI - PMC - PubMed
    1. Feneley M, Gavaghan T. Paradoxical and pseudoparadoxical interventricular septal motion in patients with right ventricular volume overload. Circulation. 1986;74:230–238. DOI: 10.1161/01.CIR.74.2.230. - DOI - PubMed

Publication types

MeSH terms

LinkOut - more resources

-