Tarasiuk A, Bar-Shimon M, Gheber L, Korngreen A, Grossman Y, Priel Z
Faculty of Natural Sciences, Department of Chemistry, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
Biophys J. 1995 Mar;68(3):1163-9. doi: 10.1016/S0006-3495(95)80292-4.
Cellular membrane potential and ciliary motility were examined in tissues cultures prepared from frog palate and esophagus epithelia. Addition of micromolar concentrations of extracellular ATP caused membrane hyperpolarization and enhanced the beat frequency. These two effects of ATP were 1) dose dependent, reaching a maximum at 10 microM ATP; 2) dependent on the presence of extracellular Ca2+ or Mg2+; 3) insensitive to inhibitors of voltage-gated calcium channels; 4) abolished after depleting the intracellular Ca2+ stores with thapsigargin; 5) attenuated by quinidine (1 mM), Cs+ (5-20 mM), and replacement of extracellular Na+ by K+; 6) insensitive to charybdotoxin (5-20 nM), TEA (1-20 microM), and apamin (0.1-1 microM); 7) independent of initial membrane potential; and 8) unaffected by amiloride. In addition, extracellular ATP induced an appreciable rise in intracellular Ca2+. Addition of thapsigargin caused an initial enhancement of the ciliary beat frequency and membrane hyperpolarization. These results strongly suggest the involvement of calcium-dependent potassium channels in the response to ATP. The results show that moderate hyperpolarization is closely associated with a sustained enhancement of ciliary beating by extracellular ATP.
在从青蛙腭和食管上皮制备的组织培养物中检测细胞膜电位和纤毛运动。添加微摩尔浓度的细胞外ATP会导致膜超极化并增加搏动频率。ATP的这两种作用为:1)剂量依赖性,在10 microM ATP时达到最大值;2)依赖于细胞外Ca2+或Mg2+的存在;3)对电压门控钙通道抑制剂不敏感;4)在用毒胡萝卜素耗尽细胞内Ca2+储存后消失;5)被奎尼丁(1 mM)、Cs+(5 - 20 mM)以及用K+替代细胞外Na+所减弱;6)对大蝎毒素(5 - 20 nM)、四乙铵(1 - 20 microM)和蜂毒明肽(0.1 - 1 microM)不敏感;7)与初始膜电位无关;8)不受氨氯吡咪影响。此外,细胞外ATP诱导细胞内Ca2+明显升高。添加毒胡萝卜素会导致纤毛搏动频率和膜超极化的初始增强。这些结果强烈表明钙依赖性钾通道参与了对ATP的反应。结果表明,适度的超极化与细胞外ATP持续增强纤毛搏动密切相关。