Acetylcholine inhibits the hypoxia-induced reduction of connexin43 protein in rat cardiomyocytes

Yanan Zhang, Yoshihiko Kakinuma, Motonori Ando, Rajesh G. Katare, Fumiyasu Yamasaki, Tetsuro Sugiura, Takayuki Sato

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36 Citations (Scopus)


In a recent study, we demonstrated that vagal stimulation increases the survival of rats with myocardial infarction by inhibiting lethal arrhythmia through regulation of connexin43 (Cx43). However, the precise mechanisms for this effect remain to be elucidated. To investigate these mechanisms and the signal transduction for gap junction regulation, we investigated the effect of acetylcholine (ACh), a parasympathetic nerve system neurotransmitter, on the gap junction component Cx43 using H9c2 cells. When cells were subjected to hypoxia, the total Cx43 protein level was decreased. In contrast, pretreatment with ACh inhibited this effect. To investigate the signal transduction, cells were pretreated with L-NAME, a nitric oxide synthase inhibitor, followed by ACh and hypoxia. L-NAME was found to suppress the ACh effect. However, a NO donor, SNAP, partially inhibited the hypoxia-induced reduction in Cx43. To delineate the mechanisms of the decrease in Cx43 under hypoxia, cells were pretreated with MG132, a proteasome inhibitor. Proteasome inhibition produced a striking recovery of the decrease in the total Cx43 protein level under hypoxia. However, cotreatment with MG132 and ACh did not produce any further increase in the total Cx43 protein level. Functional studies using ACh or okadaic acid, a phosphatase inhibitor, revealed that both reagents inhibited the decrease in the dye transfer induced by hypoxia. These results suggest that ACh is responsible for restoring the decrease in the Cx43 protein level, resulting in functional activation of gap junctions.

Original languageEnglish
Pages (from-to)214-222
Number of pages9
JournalJournal of Pharmacological Sciences
Issue number3
Publication statusPublished - 2006
Externally publishedYes


  • Acetylcholine
  • Cardiomyocyte
  • Connexin43
  • Hypoxia
  • Proteasome inhibitor

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology


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