TY - JOUR
T1 - Using Gjd3-CreEGFP mice to examine atrioventricular node morphology and composition
AU - Bhattacharyya, Samadrita
AU - Duan, Jialei
AU - Wang, Lin
AU - Li, Boxun
AU - Bhakta, Minoti
AU - Fernandez-Perez, Antonio
AU - Hon, Gary C.
AU - Munshi, Nikhil V.
N1 - Funding Information:
We thank all members of the Munshi laboratory for insightful discussions. This work was supported by the AHA (17PRE33670730 to S.B.), NIH (HL136604, HL133642, and HL135217 to N.V.M; DP2GM128203 to G.C.H.), the Burroughs Wellcome Fund (1009838 to N.V.M.), the March of Dimes Foundation (#5-FY13-203 to N.V.M.), the Department of Defense (PR172060 to G.C.H. and N.V.M.), the Cancer Prevention Research Institute of Texas (CPRIT) (RR140023 to G.C.H.), the Welch Foundation (I-1926-20170325 to G.C.H.), and the Green Center for Reproductive Biology. We acknowledge John Shelton and the Histology Core for providing lacZ-stained sections, the McDermott Center for next-generation sequencing, and the BioHPC computational infrastructure for providing HPC and storage resources.
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - The atrioventricular node (AVN) coordinates the timing of atrial and ventricular contraction to optimize cardiac performance. To study this critical function using mouse genetics, however, new reagents are needed that allow AVN-specific manipulation. Here we describe a novel Gjd3-CreEGFP mouse line that successfully recombines floxed alleles within the AVN beginning at E12.5. These mice have been engineered to express CreEGFP under the control of endogenous Gjd3 regulatory elements without perturbing native protein expression. Detailed histological analysis of Gjd3-CreEGFP mice reveals specific labeling of AVN cardiomyocytes and a subset of cardiac endothelial cells. Importantly, we show that Gjd3-CreEGFP mice have preserved cardiac mechanical and electrical function. In one application of our newly described mouse line, we provide a three-dimensional (3D) view of the AVN using tissue clearing combined with confocal microscopy. With this 3D model as a reference, we identify specific AVN sub-structures based on marker staining characteristics. In addition, we use our Gjd3-CreEGFP mice to guide microdissection of the AVN and construction of a single-cell atlas. Thus, our results establish a new transgenic tool for AVN-specific recombination, provide an updated model of AVN morphology, and describe a roadmap for exploring AVN cellular heterogeneity.
AB - The atrioventricular node (AVN) coordinates the timing of atrial and ventricular contraction to optimize cardiac performance. To study this critical function using mouse genetics, however, new reagents are needed that allow AVN-specific manipulation. Here we describe a novel Gjd3-CreEGFP mouse line that successfully recombines floxed alleles within the AVN beginning at E12.5. These mice have been engineered to express CreEGFP under the control of endogenous Gjd3 regulatory elements without perturbing native protein expression. Detailed histological analysis of Gjd3-CreEGFP mice reveals specific labeling of AVN cardiomyocytes and a subset of cardiac endothelial cells. Importantly, we show that Gjd3-CreEGFP mice have preserved cardiac mechanical and electrical function. In one application of our newly described mouse line, we provide a three-dimensional (3D) view of the AVN using tissue clearing combined with confocal microscopy. With this 3D model as a reference, we identify specific AVN sub-structures based on marker staining characteristics. In addition, we use our Gjd3-CreEGFP mice to guide microdissection of the AVN and construction of a single-cell atlas. Thus, our results establish a new transgenic tool for AVN-specific recombination, provide an updated model of AVN morphology, and describe a roadmap for exploring AVN cellular heterogeneity.
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U2 - 10.1038/s41598-019-38683-8
DO - 10.1038/s41598-019-38683-8
M3 - Article
C2 - 30765799
AN - SCOPUS:85061596911
VL - 9
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
IS - 1
M1 - 2106
ER -