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Sci Rep. 2019 Jul 24;9(1):10716. doi: 10.1038/s41598-019-47009-7.

Simplified platform for mosaic in vivo analysis of cellular maturation in the developing heart.

Author information

1
Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, USA.
2
McAllister Heart Institute, University of North Carolina at Chapel Hill, Chapel Hill, USA.
3
Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, USA. Michael_Bressan@med.unc.edu.
4
McAllister Heart Institute, University of North Carolina at Chapel Hill, Chapel Hill, USA. Michael_Bressan@med.unc.edu.

Abstract

Cardiac cells develop within an elaborate electro-mechanical syncytium that continuously generates and reacts to biophysical force. The complexity of the cellular interactions, hemodynamic stresses, and electrical circuitry within the forming heart present significant challenges for mechanistic research into the cellular dynamics of cardiomyocyte maturation. Simply stated, it is prohibitively difficult to replicate the native electro-mechanical cardiac microenvironment in tissue culture systems favorable to high-resolution cellular/subcellular analysis, and current transgenic models of higher vertebrate heart development are limited in their ability to manipulate and assay the behavior of individual cells. As such, cardiac research currently lacks a simple experimental platform for real-time evaluation of cellular function under conditions that replicate native development. Here we report the design and validation of a rapid, low-cost system for stable in vivo somatic transgenesis that allows for individual cells to be genetically manipulated, tracked, and examined at subcellular resolution within the forming four-chambered heart. This experimental platform has several advantages over current technologies, chief among these being that mosaic cellular perturbations can be conducted without globally altering cardiac function. Consequently, direct analysis of cellular behavior can be interrogated in the absence of the organ level adaptions that often confound data interpretation in germline transgenic model organisms.

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