- Poster presentation
- Open Access
- Published:
Micro-MRI phenotyping of a novel double-knockout mouse model of congenital heart disease
Journal of Cardiovascular Magnetic Resonance volume 12, Article number: P1 (2010)
Introduction
CHARGE and DiGeorge syndromes are conditions with incidences of 1 in 10,000 and 1 in 4000 and are strongly associated with haploinsufficiency of specific genes (CHD7 and TBX1). Both conditions are characterised by cardiovascular defects. Knockout mouse models are an important tool for the identification and characterisation of genes implicated in congenital heart conditions. Micro-MRI is an emerging technique for high resolution cardiac phenotyping in a reduced time compared to conventional histology, enabling the acquisition of 3D images of multiple embryos in a single scan[1].
Purpose
Given the phenotypic overlap of these conditions we sought to examine the effect on cardiac morphology in double-knockout mouse embryos (Chd7+/-Tbx1+/-)[2], performing an initial assessment of these mice using MRI.
Methods
Study Design
18 embryos (1 wild-type, 7 Chd7+/-, 2 Tbx1+/- and 8 Chd7+/-Tbx1+/-) were imaged and examined for cardiac abnormalities.
Embryo Preparation
16.5 dpc embryos were fixed for at least 2 weeks in a solution of 4% formaldehyde-PBS with 8 mM Gd-DTPA (Bayer-Schering AG) and then embedded in 1% agarose gel (doped with 8 mM Gd-DTPA) in 50 ml centrifuge tubes.
Imaging
Performed on a Varian 9.4 T VNMRS system with 33 mm quadrature birdcage coil (RAPID Biomedical GmbH), using a 3D gradient echo sequence (TE/TR/FA/NSA = 9/20/60/7, FOV = 27 × 27 × 27 mm3, voxel size = 52 × 52 × 52 μm3).
Image analysis
Datasets were zero-filled to 26 × 26 × 26 μm3 and reviewed in Amira (v5.2, Visage Imaging Inc.).
Histology
Embryos identified with abnormal hearts by MRI, were then histologically examined by H&E staining.
Results
Of the 18 embryos scanned for MR analysis, we identified abnormal (thin or patent) ventricular septa (VSDs) in 6 embryos. Of these, 1 was a Chd7+/-heterozygote (an incidence of 1/7 in this study, see Fig. 1). VSDs were seen in 5/8 Chd7+/-Tbx1 embryos (Fig. 2). In one double heterozygote we also observed an interrupted aortic arch, in addition to a VSD, which was readily seen by MRI (Fig. 3).
Conclusion
Using micro-MRI we have successfully identified cardiac abnormalities in genetically-modified embryos. A single Chd7+/- embryo was found to have an abnormal ventricular septum. A relatively high incidence of VSDs was observed in Chd7+/-Tbx1+/- compared to Chd7+/-mice, indicating possible interaction of these two genes. An interrupted aortic arch was identified in one Chd7+/-Tbx1+/- embryo. All abnormal findings were later confirmed by histology which indicates that micro-MRI is an effective technique for cardiac phenotyping.
References
Cleary JO, Price AN, et al.: NMR Biomed. 2009, 22 (8): 857-866. 10.1002/nbm.1400.
McCue K, Randall V, et al.: 06-P038. Mech of Dev. 2009, 126: S131-10.1016/j.mod.2009.06.264.
Author information
Authors and Affiliations
Rights and permissions
Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/2.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
About this article
Cite this article
Cleary, J.O., McCue, K., Price, A.N. et al. Micro-MRI phenotyping of a novel double-knockout mouse model of congenital heart disease. J Cardiovasc Magn Reson 12 (Suppl 1), P1 (2010). https://doi.org/10.1186/1532-429X-12-S1-P1
Published:
DOI: https://doi.org/10.1186/1532-429X-12-S1-P1
Keywords
- Congenital Heart Disease
- Cardiac Abnormality
- Ventricular Septum
- Knockout Mouse Model
- DiGeorge Syndrome