Otsuguro K, Ohta T, Ito S, Nakazato Y
Laboratory of Pharmacology, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060, Japan.
Pflugers Arch. 1996 Jan;431(3):402-7. doi: 10.1007/BF02207278.
The effects of extracellular adenosine 5'-triphosphate (ATP) on voltage-dependent Ca2+ currents were examined using the whole-cell voltage-clamp technique in guinea-pig isolated adrenal chromaffin cells. ATP (500 microM) reversibly suppressed Ca2+ currents in the presence of 5mM Ca2+ in the extracellular solution. The inhibitory effect of ATP on Ca2+ currents tended to increase with increases in the peak amplitude of ATP-evoked current when the intracellular solution contained 0.1 or 1 mM ethylenebis(oxonitrilo)tetraacetate(EGTA). Using the intracellular solution containing 10mM EGTA, on the other hand, the inhibitory effect did not change regardless of the amplitude of current responses to ATP. In the presence of 10 mM Ba2+, ATP (100 microM) reduced Ba2+ currents in a manner similar to Ca2+ currents. This reduction was decreased by dialysis of cells with the internal solution containing guanosine 5'-O-(2-thiodiphosphate) (GDP [beta-S]; 1 mM) or guanosine 5'-O-(3-thiotriphosphate) (GTP [gamma-S]; 100 microM). A depolarizing prepulse to + 100 mV partly relieved ATP-induced reduction of Ba2+ currents. ADP, AMP and adenosine also reduced Ba2+ currents and the effect of adenosine was the most potent. Adenosine (0.5 and 1 mM) significantly inhibited adrenaline secretion induced by nicotine (50 microM). These results suggest that Ca2+ entry through ATP-activated non-selective cation channels results in the inactivation of voltage-dependent Ca2+ channels. In addition, ATP seems to modulate Ca2+ channels via the pathway related to G-protein. Adenine nucleotides and adenosine may play a role in controlling secretory activity in guinea-pig adrenal chromaffin cells.
采用全细胞膜片钳技术,在豚鼠分离的肾上腺嗜铬细胞中研究了细胞外5'-三磷酸腺苷(ATP)对电压依赖性Ca2+电流的影响。在细胞外溶液中存在5 mM Ca2+的情况下,ATP(500 μM)可逆性抑制Ca2+电流。当细胞内溶液含有0.1或1 mM乙二胺四乙酸(EGTA)时,ATP对Ca2+电流的抑制作用倾向于随着ATP诱发电流峰值幅度的增加而增强。另一方面,使用含有10 mM EGTA的细胞内溶液时,无论对ATP的电流反应幅度如何,抑制作用均无变化。在存在10 mM Ba2+的情况下,ATP(100 μM)以类似于Ca2+电流的方式降低Ba2+电流。用含有5'-O-(2-硫代二磷酸)鸟苷(GDP[β-S];1 mM)或5'-O-(3-硫代三磷酸)鸟苷(GTP[γ-S];100 μM)的内部溶液对细胞进行透析可减弱这种降低作用。向+100 mV的去极化预脉冲可部分缓解ATP诱导的Ba2+电流降低。二磷酸腺苷(ADP)、一磷酸腺苷(AMP)和腺苷也可降低Ba2+电流,其中腺苷的作用最为显著。腺苷(0.5和1 mM)可显著抑制尼古丁(50 μM)诱导的肾上腺素分泌。这些结果表明,通过ATP激活的非选择性阳离子通道进入的Ca2+导致电压依赖性Ca2+通道失活。此外,ATP似乎通过与G蛋白相关的途径调节Ca2+通道。腺嘌呤核苷酸和腺苷可能在豚鼠肾上腺嗜铬细胞分泌活性的控制中发挥作用。