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Increase of L-type Calcium Current by cGMP-dependent Protein Kinase Regulates in Rabbit Ventricular Myocytes

Han, Jin   (Department of Physiology and Biophysics, College of Medicine, Inje UniversityUU0001067  ); Kim, Na-Ri   (Department of Physiology and Biophysics, College of Medicine, Inje UniversityUU0001067  ); Kim, Eui-Yong   (Department of Physiology and Biophysics, College of Medicine, Inje UniversityUU0001067  ); Ho, Won-Kyung   (Department of Physiology and Biophysics, College of Medicine, Seoul National UniversityUU0000691  ); Earm, Yung-E   (Department of Physiology and Biophysics, College of Medicine, Seoul National UniversityUU0000691  ); Kim, Han-Kyoun   (Department of Orthopedic Surgery, Bohoon HospitalHH0000120  );
  • 초록

    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.


  • 주제어

    cGMP-dependent protein kinase (cGMP-PK) .   L-type calcium current $(I_{Ca})$ .   Rabbit ventricular myocytes .   Whole-cell voltage clamp.  

 저자의 다른 논문

  • Han, Jin (34)

    1. 1992 "A Study on the $Na^+/Ca^{2+}$ Exchange Mechanism in the Smooth Muscle of Guinea-pig Stomach" 대한생리학회지 = Korean journal of physiology 26 (1): 55~68    
    2. 1992 "Calcium Current and Background Current Activation in L-triiodothyronine Loaded Ventricular Myocytes of the Rabbit" 대한생리학회지 = Korean journal of physiology 26 (2): 99~111    
    3. 1992 "Effects of $Cd^{2+}$ on the Contractility in the Antral Circular Muscle of Guinea-pig Stomach" 대한생리학회지 = Korean journal of physiology 26 (2): 129~136    
    4. 1993 "Effect of Cyclic GMP on the Calcium Current in Rabbit Ventricular Myocytes" 대한생리학회지 = Korean journal of physiology 27 (2): 151~162    
    5. 1994 "Role of $Na^+/Ca^{2+}$ Exchange in the Control of Contractility in Rabbit Basilar Arterial Smooth Muscle" 대한생리학회지 = Korean journal of physiology 28 (2): 159~167    
    6. 1996 "Effects of Adenosine on the Ionic Channel Activated by Metabolic Inhibition in Rabbit Ventricular Myocytes" 대한생리학회지 = Korean journal of physiology 30 (1): 1~9    
    7. 1998 "The Effects of Intracellular Monocarboxylates on the ATP-sensitive Potassium Channels in Rabbit Ventricular Myocytes" The Korean journal of physiology & pharmacology : official journal of the Korean Physiological Society and the Korean Society of Pharmacology 2 (5): 581~589    
    8. 1999 "Role of $K^+$ Channels to Resting Membrane Potential of Rabbit Middle Cerebral Arterial Smooth Muscle Cells" The Korean journal of physiology & pharmacology : official journal of the Korean Physiological Society and the Korean Society of Pharmacology 3 (6): 547~554    
    9. 2000 "Effects of Prostaglandin $F_{2{\alpha}}$ on Membrane Potentials and $K^+$ Currents in Rabbit Middle Cerebral Arterial Cells" The Korean journal of physiology & pharmacology : official journal of the Korean Physiological Society and the Korean Society of Pharmacology 4 (4): 301~309    
    10. 2000 "Modulation of $Ca^{2+}-Activated$ Potassium Channels by cGMP-Dependent Signal Transduction Mechanism in Cerebral Arterial Smooth Muscle Cell of the Rabbit" The Korean journal of physiology & pharmacology : official journal of the Korean Physiological Society and the Korean Society of Pharmacology 4 (6): 445~453    
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