Background-We recently reported that cardiomyocytes could be differentiated from bone marrow mesenchymal stem cells in vitro by 5-azacytidine treatment. In native cardiomyocytes, adrenergic and muscarinic receptors play crucial roles in mediating heart rate, conduction velocity, contractility, and cardiac hypertrophy. We investigated whether these receptors are expressed in differentiated CMG cells, and if so, whether they have downstream signaling systems. Methods and Results-Reverse transcription-polymerase chain reaction revealed that CMG cells had already expressed ␣ 1A -, ␣ 1B -, and ␣ 1D -adrenergic receptor mRNA before 5-azacytidine treatment, whereas expression of  1 -,  2 -adrenergic and M 1 -, M 2 -muscarinic receptors was first detected at 1 day. Phenylephrine dose-dependently induced phosphorylation of ERK1/2, which was completely inhibited by prazosin, and significantly increased cell size. Isoproterenol augmented cAMP by 38-fold, which was fully inhibited by propranolol. Isoproterenol (10 Ϫ7 mol/L) increased the spontaneous beating rate by 47.6% (basal, 127Ϯ16 bpm), and propranolol and CGP20712A ( 1 -selective blocker) reduced it by 79.0% and 71.0%, respectively, whereas ICI118551 ( 2 -selective blocker) induced slight reduction. Cell motion, percent shortening, and contractile velocity were increased by 37.5%, 26.9%, and 50.6%, respectively, in response to isoproterenol. Phenylephrine and isoproterenol augmented ANP and BNP gene expressions. Carbachol increased IP 3 by 32-fold, which was markedly inhibited by atropine as well as AFDX116 (M 2 -selective blocker) measured by radioimmunoassay.
Conclusions-These