科研之友
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摘要
Studies in isolated cardiomyocytes have provided tremendous information at the cellular and molecular level concerning regulation of transmembrane voltage (V-m) and intracellular calcium ([Ca2+](i)). The ability to use the information gleaned to gain insight into the function of ion channels and Ca2+ handling proteins in a more complex system, e.g., the intact heart, has remained a challenge. We have developed laser scanning fluorescence microscopy-based approaches to monitor, at the sub-cellular to multi-cellular level in the immobilized, Langendorff-perfused mouse heart, dynamic changes in [Ca2+](i) and V-m. This article will review the use of single- or dual-photon laser scanning microscopy [Ca2+](i) imaging in conjunction with transgenic reporter technology to (a) interrogate the extent to which transplanted, donor-derived myocytes or cardiac stem cell-derived de novo myocytes are capable of forming a functional syncytium with the pre-existing myocardium, using entrainment of [Ca2+](i) transients by the electrical activity of the recipient heart as a surrogate for electrical coupling, and (b) characterize the Ca2+ handling phenotypes of cellular implants. Further, we will review the ability of laser scanning fluorescence microscopy in conjunction with a fast-response voltage-sensitive to resolve, on a subcellular level in Langendorff-perfused mouse hearts, V-m dynamics that typically occur during the course of a cardiac action potential. Specifically, the utility of this technique to measure microscopic-scale voltage gradients in the normal and diseased heart is discussed.
- 出版日期2014-12-2
