Abstract
Connexin43 (Cx43) is the predominant intercellular gap junction protein in the heart providing intercellular communication for the cell-to-cell transfer of electrical activation. In a previous study, we could show that alpha-adrenoceptor stimulation can affect Cx43 expression and function. We now wanted to elucidate which alpha1-adrenoceptor subtype might be involved. Cultured neonatal rat cardiomyocytes were exposed to various concentrations of phenylephrine (0.1-1,000 nM) for 24 h (n=6). Thereafter, cells were harvested, and after lysis, Cx43 content was determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot. Results were normalised to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Finally, we determined the effect of this treatment on intercellular gap junction conductivity using dual whole-cell voltage clamp. Similarly, we tested the effect of an additional treatment with either 10 nM prazosin or, to assess the subtypes, 10 nM of the alpha(1A)-antagonist RS17053 (n=6), 500 nM of the alpha(1B)-antagonist AH1111OA (n=6), or 50 nM of the alpha(1D)-antagonist BMY7378 (n = 6). Moreover, we incubated the cells for 24 h with the alpha(1A)-adrenoceptor agonist A61603 (10 nM). Phenylephrine le...Continue Reading
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