Electrophysiological disorders of the heart by Sanjeev Saksena; A John Camm; et al

By Sanjeev Saksena; A John Camm; et al

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For example, the N-terminal domain of cardiac TnI is phosphorylated by cyclic adenosine monophosphate (cAMP)-dependent PKA phosphorylation at serines 22 and 23. 8 Thus, these findings clearly illustrate the highly collaborative roles of structural, electrical, mechanical, and signaling proteins in the modulation of myocyte EC coupling and cardiac function. Recent findings demonstrate key cellular roles for nuclear and mitochondrial membranes in the regulation of myocyte transcriptional pathways and in metabolism.

Gap junctions in ventricular tissue consist primarily of connexin43, which forms large conductance channels to allow rapid conduction. In contrast, gap junctions in the sinus node contain mostly connexin45, which forms lower conductance channels ideal for slow but safe conduction. The adherens junction is maintained by the function of a cadherin-catenin complex that provides a stabilizing link from the intercalated disc to the actin cytoskeleton (see Figure 2-3). Finally, the desmosome also supports cell adhesion through a complex, including plakoglobin (γ-catenin, also found in the adherens junction), desmoplakin, and plakophilin, that interacts with intermediate filaments (see Figure 2-3).

Commonly used cell lines include mouse fibroblasts (L cells), human embryonic kidney cells (HEK293), SV40immortalized, African green monkey (Cos) cells, and Chinese hamster ovary (CHO) cells. These cell lines are easily grown in culture, are readily transfectable, are capable of high-level, exogenous protein expression, and have relatively few endogenous currents. Stable transfection of wild-type or mutant channels can be achieved by introducing antibiotic selection. Known mutations from human channelopathies can be propagated in stable cell lines, allowing detailed electrophysiologic analysis.

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