Inhibition of connexin 43 in cardiac muscle during intense physical exercise.

Endurance training is accompanied by important adaptations in both cardiovascular and autonomic nervous systems. Previous works have shown that the main component of gap junctions in the ventricular myocardium (connexin 43 (Cx43) can be regulated by adrenergic stimulus. On the other hand, training raises vagal and decreases sympathetic tone, while augmenting myocardial sensitivity to sympathetic stimulation during exercise. We therefore evaluated the regulation of Cx43 expression by sympathetic tone during exercise in trained and sedentary mice. Training induced an increase in the protein level of Cx43 by 45-70% under resting conditions. The expression of Cx43 was inhibited in trained but not in untrained mice in response to a 60 min exercise bout. Normal basal expression was restored after 60 min of resting. Cx43 reached a minimum that was not different from basal expression in untrained mice. In accordance, electrocardiography and action potential analysis did not reveal major electrophysiological implications for the drop in Cx43 abundance in trained-exercise mice. We prevented Cx43 inhibition using propranolol, and observed increased basal mRNA levels of ß-adrenergic receptors without significant changes in the ratio ß1 to ß2. In conclusion, we showed that Cx43 expression is transiently inhibited by ß-adrenergic stimulus in trained mice during acute exercise.

Glucocorticoid-mediated destabilization of cyclin D3 mRNA involves RNA-protein interactions in the 3'-untranslated region of the mRNA.

Glucocorticoids regulate the expression of the G(1) progression factor, cyclin D3. Cyclin D3 messenger RNA (CcnD3 mRNA) stability decreases rapidly when murine T lymphoma cells are treated with the synthetic glucocorticoid dexamethasone. Basal stability of CcnD3 mRNA is regulated by sequences within the 3'-untranslated region (3'-UTR). RNA-protein interactions occurring within the CcnD3 3'-UTR have been analyzed by RNA electrophoretic mobility shift assay. Three sites of RNA-protein interaction have been mapped using this approach. These elements include three pyrimidine-rich domains of 25, 26, and 37 nucleotides. When the cyclin D3 3'-UTR was stably overexpressed, the endogenous CcnD3 mRNA was no longer regulated by dexamethasone. Likewise, overexpression of a 215-nucleotide transgene that contains the 26- and 37-nucleotide elements blocks glucocorticoid inhibition of CcnD3 mRNA expression. These observations suggest that the 215-nucleotide 3'-UTR element may act as a molecular decoy, competing for proteins that bind to the endogenous transcript and thereby attenuating glucocorticoid responsiveness. UV-cross-linking experiments showed that two proteins of approximate molecular weight 37,000 and 52,000 bind to this 3'-UTR element.