Regulation of protein biosynthesis in neonatal rat cardiomyocytes by adrenoceptor-stimulation: investigations with high resolution two-dimensional polyacrylamide gel electrophoresis.
The role of alpha- and beta-adrenoceptor-mediated regulation of protein patterns in cultured neonatal rat cardiomyocytes was studied with high resolution two-dimensional polyacrylamide gel electrophoresis. Spontaneously beating neonatal rat cardiomyocytes were cultured for 72 h at 37 degrees C in serum-free medium, supplemented with either the catecholamine norepinephrine (0.1 microM), or with norepinephrine (0.1 microM) plus the alpha 1-adrenoceptor blocker prazosin (1 microM), or with neither of these substances. In transmission light, 105 protein spots could be seen in the lysates from control cells, 244 spots were identified in lysates from norepinephrine-treated cells, and 114 protein species were counted in the case of lysates from cardiomyocytes which had been cultured in the presence of norepinephrine plus prazosin. This experimental approach allows a clear classification of alpha- and non-alpha (probably beta)-adrenoceptor-mediated catecholamine effects on protein synthesis in cardiomyocytes. In comparison with previous experiments (Chang et al., Electrophoresis, this issue, pp. 748-754), less protein species were identified in the untreated control cardiomyocytes, as well as in the norepinephrine treated cells. The only actual modification in the experimental setup in these two series was the addition of dimethyl sulfoxide--a substance known for its repressive effect on oncoprotein expression--to the culture medium, as solvent for prazosin.
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Department of Medicine I, University of Munich, Klinikum Grosshadern, Germany.
Müller
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Werdan
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D000818:Animals; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006321:Heart / Q000187:drug effects; D009206:Myocardium / Q000378:metabolism*; D009638:Norepinephrine / Q000494:pharmacology; D010449:Peptide Mapping / Q000379:methods; D011224:Prazosin / Q000494:pharmacology; D014176:Protein Biosynthesis; D011506:Proteins / Q000302:isolation & purification; D051381:Rats; D011941:Receptors, Adrenergic / Q000187:drug effects / Q000378:metabolism*
D011506:Proteins; D011941:Receptors, Adrenergic; D009638:Norepinephrine; D011224:Prazosin