Rosário L M, Barbosa R M, Antunes C M, Silva A M, Abrunhosa A J, Santos R M
Departamento de Bioquímica, Faculdade de Ciências e Tecnologia da Universidade de Coimbra, Portugal.
Pflugers Arch. 1993 Sep;424(5-6):439-47. doi: 10.1007/BF00374906.
In glucose-stimulated pancreatic beta-cells, the membrane potential alternates between a hyperpolarized silent phase and a depolarized phase with Ca2+ action potentials. The molecular and ionic mechanisms underlying these bursts of electrical activity remain unknown. We have observed that 10.2-12.8 mM Ca2+, 1 microM Bay K 8644 and 2 mM tetraethylammonium (TEA) trigger bursts of electrical activity and oscillations of intracellular free Ca2+ concentration ([Ca2+]i) in the presence of 100 microM tolbutamide. The [Ca2+]i was monitored from single islets of Langerhans using fura-2 microfluorescence techniques. Both the high-Ca(2+)- and Bay-K-8644-evoked [Ca2+]i oscillations overshot the [Ca2+]i recorded in tolbutamide. Nifedipine (10-20 microM) caused an immediate membrane hyperpolarization, which was followed by a slow depolarization to a level close to the burst active phase potential. The latter depolarization was accompanied by suppression of spiking activity. Exposure to high Ca2+ in the presence of nifedipine caused a steady depolarization of approximately 8 mV. Ionomycin (10 microM) caused membrane hyperpolarization in the presence of 7.7 mM Ca2+, which was not abolished by nifedipine. Charybdotoxin (CTX, 40-80 nM), TEA (2 mM) and quinine (200 microM) did not suppress the high-Ca(2+)-evoked bursts. It is concluded that: (1) the channel underlying the burst is sensitive to [Ca2+]i rises mediated by Ca2+ influx through L-type Ca2+ channels, (2) both the ATP-dependent K+ channel and the CTX- and TEA-sensitive Ca(2+)-dependent K+ channel are highly unlikely to provide the pacemaker current underlying the burst.(ABSTRACT TRUNCATED AT 250 WORDS)
在葡萄糖刺激的胰腺β细胞中,膜电位在超极化静息期和伴有Ca2+动作电位的去极化期之间交替。这些电活动爆发背后的分子和离子机制仍然未知。我们观察到,在存在100μM甲苯磺丁脲的情况下,10.2 - 12.8 mM Ca2+、1μM Bay K 8644和2 mM四乙铵(TEA)会引发电活动爆发以及细胞内游离Ca2+浓度([Ca2+]i)的振荡。使用fura - 2微荧光技术从单个胰岛监测[Ca2+]i。高Ca2+和Bay - K - 8644诱发的[Ca2+]i振荡均超过了在甲苯磺丁脲中记录的[Ca2+]i。硝苯地平(10 - 20μM)引起立即的膜超极化,随后缓慢去极化至接近爆发活跃期电位的水平。后者的去极化伴随着尖峰活动的抑制。在硝苯地平存在的情况下暴露于高Ca2+会导致约8 mV的稳定去极化。离子霉素(10μM)在存在7.7 mM Ca2+的情况下引起膜超极化,硝苯地平不能消除这种超极化。蝎毒素(CTX,40 - 80 nM)、TEA(2 mM)和奎宁(200μM)不会抑制高Ca2+诱发的爆发。得出以下结论:(1)爆发背后的通道对通过L型Ca2+通道的Ca2+内流介导的[Ca2+]i升高敏感;(2)ATP依赖性K+通道以及对CTX和TEA敏感的Ca2+依赖性K+通道极不可能提供爆发背后的起搏电流。(摘要截短于250字)