Increase of L-type calcium current by cGMP-dependent protein kinase regulates in rabbit ventricular myocytes

J. Han, N. Kim, E. Kim, Wonkyung Ho, Y. E. Earm, H. Kim

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Abstract

Background: We have previously reported that not only cGMP but also 8- Br-cGMP or 8-pCPT-cGMP, specific and potent stimulators of cGMP-dependent protein kinase (cGMP-PK), increased basal L-type calcium current (I(Ca)) in rabbit ventricular myocytes. Our findings in rabbit ventricular myocytes were entirely different from the earlier findings in different species, suggesting that the activation of cGMP-PK is involved in the facilitation of I(Ca) by cGMP. However, there is no direct evidence that cGMP-PK can stimulate I(Ca) in rabbit ventricular myocytes. In this report, we focused on the direct effect of cGMP-PK on I(Ca) in rabbit ventricular myocytes. Methods and Results: We isolated single ventricular myocytes of rabbit hearts by using enzymatic dissociation. Regulation of I(Ca) by cGMP-PK was investigated in rabbit ventricular myocytes using whole-cell voltage clamp method. I(Ca) was elicited by a depolarizing pulse to +10 mV from a holding potential of -40 mV. Extracellular 8-(4-Chlorophenylthio)-guanosine-3',5'-cyclic monophosphate (8-pCPT-cGMP), potent stimulator of cGMP-dependent protein kinase (cGMP-PK), increased basal I(Ca). cGMP-PK also increased basal I(Ca). The stimulation of basal I(Ca) by cGMP-PK required both 8-Br-cGMP in low concentration and intracellular ATP to be present. The stimulation of basal I(Ca) by cGMP-PK was blocked by heat inactivation of the cGMP-PK and by bath application of 8(4-chlorophenylthio)-guanosine-3',5'-cyclic monophosphate, Rp-isomer (Rp- pCPT-cGMP), a phosphodiesterase-resistant cGMP-PK inhibitor. When I(Ca) was increased by internal application of cGMP-PK, IBMX resulted in an additional stimulation of I(Ca). In the presence of cGMP-PK, already increased I(Ca) was potentiated by bath application of isoprenaline or forskolin or intracellular application of cAMP. Conclusions: We present evidence that cGMP-PK stimulated basal I(Ca) by a direct phosphorylation of L-type calcium channel or associated regulatory protein in rabbit ventricular myocytes.

Original languageEnglish
Pages (from-to)733-742
Number of pages10
JournalKorean Journal of Physiology and Pharmacology
Volume2
Issue number6
StatePublished - 1 Dec 1998

Keywords

  • L-type calcium current (I(Ca))
  • Rabbit ventricular myocytes
  • Whole-cell voltage clamp
  • cGMP-dependent protein kinase (cGMP-PK)

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