Bone marrow-derived regenerated cardiomyocytes (CMG cells) express functional adrenergic and muscarinic receptors

Daihiko Hakuno, Keiichi Fukuda, Shinji Makino, Fusako Konishi, Yuichi Tomita, Tomohiro Manabe, Yusuke Suzuki, Akihiro Umezawa, Satoshi Ogawa

Research output: Contribution to journalArticle

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Abstract

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 α1A-, α1B-, and α1D-adrenergic receptor mRNA before 5-azacytidine treatment, whereas expression β1-, β2-adrenergic and M1-, M2-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 (10-7 mol/L) increased the spontaneous beating rate by 47.6% (basal, 127 ± 16 bpm), and propranolol and CGP20712A (β1-selective blocker) reduced it by 79.0% and 71.0%, respectively, whereas ICI118551 (β2-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 IP3 by 32-fold, which was markedly inhibited by atropine as well as AFDX116 (M2-selective blocker) measured by radioimmunoassay. Conclusions - These findings indicate that CMG cells expressed α1A, α1B, and α1D receptors before differentiation and expressed β1, β2, M1, and M2 receptors after they obtained the cardiomyocyte phenotype. These receptors had functional signal transduction pathways and could modulate cell function.

Original languageEnglish
Pages (from-to)380-386
Number of pages7
JournalCirculation
Volume105
Issue number3
DOIs
Publication statusPublished - 2002 Jan 22

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Muscarinic Receptors
Cardiac Myocytes
Adrenergic Receptors
Isoproterenol
Bone Marrow
Azacitidine
Phenylephrine
Propranolol
Muscarinic M2 Receptors
Muscarinic M1 Receptors
Prazosin
Cardiomegaly
Carbachol
Atrial Natriuretic Factor
Mesenchymal Stromal Cells
Atropine
Cell Size
Adrenergic Agents
Reverse Transcription
Radioimmunoassay

Keywords

  • Cells
  • Heart rate
  • Receptors, adrenergic, alpha
  • Receptors, adrenergic, beta
  • Signal transduction

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Bone marrow-derived regenerated cardiomyocytes (CMG cells) express functional adrenergic and muscarinic receptors. / Hakuno, Daihiko; Fukuda, Keiichi; Makino, Shinji; Konishi, Fusako; Tomita, Yuichi; Manabe, Tomohiro; Suzuki, Yusuke; Umezawa, Akihiro; Ogawa, Satoshi.

In: Circulation, Vol. 105, No. 3, 22.01.2002, p. 380-386.

Research output: Contribution to journalArticle

Hakuno, Daihiko ; Fukuda, Keiichi ; Makino, Shinji ; Konishi, Fusako ; Tomita, Yuichi ; Manabe, Tomohiro ; Suzuki, Yusuke ; Umezawa, Akihiro ; Ogawa, Satoshi. / Bone marrow-derived regenerated cardiomyocytes (CMG cells) express functional adrenergic and muscarinic receptors. In: Circulation. 2002 ; Vol. 105, No. 3. pp. 380-386.
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abstract = "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 α1A-, α1B-, and α1D-adrenergic receptor mRNA before 5-azacytidine treatment, whereas expression β1-, β2-adrenergic and M1-, M2-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 (10-7 mol/L) increased the spontaneous beating rate by 47.6{\%} (basal, 127 ± 16 bpm), and propranolol and CGP20712A (β1-selective blocker) reduced it by 79.0{\%} and 71.0{\%}, respectively, whereas ICI118551 (β2-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 IP3 by 32-fold, which was markedly inhibited by atropine as well as AFDX116 (M2-selective blocker) measured by radioimmunoassay. Conclusions - These findings indicate that CMG cells expressed α1A, α1B, and α1D receptors before differentiation and expressed β1, β2, M1, and M2 receptors after they obtained the cardiomyocyte phenotype. These receptors had functional signal transduction pathways and could modulate cell function.",
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T1 - Bone marrow-derived regenerated cardiomyocytes (CMG cells) express functional adrenergic and muscarinic receptors

AU - Hakuno, Daihiko

AU - Fukuda, Keiichi

AU - Makino, Shinji

AU - Konishi, Fusako

AU - Tomita, Yuichi

AU - Manabe, Tomohiro

AU - Suzuki, Yusuke

AU - Umezawa, Akihiro

AU - Ogawa, Satoshi

PY - 2002/1/22

Y1 - 2002/1/22

N2 - 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 α1A-, α1B-, and α1D-adrenergic receptor mRNA before 5-azacytidine treatment, whereas expression β1-, β2-adrenergic and M1-, M2-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 (10-7 mol/L) increased the spontaneous beating rate by 47.6% (basal, 127 ± 16 bpm), and propranolol and CGP20712A (β1-selective blocker) reduced it by 79.0% and 71.0%, respectively, whereas ICI118551 (β2-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 IP3 by 32-fold, which was markedly inhibited by atropine as well as AFDX116 (M2-selective blocker) measured by radioimmunoassay. Conclusions - These findings indicate that CMG cells expressed α1A, α1B, and α1D receptors before differentiation and expressed β1, β2, M1, and M2 receptors after they obtained the cardiomyocyte phenotype. These receptors had functional signal transduction pathways and could modulate cell function.

AB - 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 α1A-, α1B-, and α1D-adrenergic receptor mRNA before 5-azacytidine treatment, whereas expression β1-, β2-adrenergic and M1-, M2-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 (10-7 mol/L) increased the spontaneous beating rate by 47.6% (basal, 127 ± 16 bpm), and propranolol and CGP20712A (β1-selective blocker) reduced it by 79.0% and 71.0%, respectively, whereas ICI118551 (β2-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 IP3 by 32-fold, which was markedly inhibited by atropine as well as AFDX116 (M2-selective blocker) measured by radioimmunoassay. Conclusions - These findings indicate that CMG cells expressed α1A, α1B, and α1D receptors before differentiation and expressed β1, β2, M1, and M2 receptors after they obtained the cardiomyocyte phenotype. These receptors had functional signal transduction pathways and could modulate cell function.

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KW - Receptors, adrenergic, alpha

KW - Receptors, adrenergic, beta

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