Göpel S O, Kanno T, Barg S, Rorsman P
Department of Molecular and Cellular Physiology, Diabetes Research Unit, Institute of Physiological Sciences, Lund University, Solvegatan 19, S-223 62 Lund, Sweden.
J Physiol. 2000 Nov 1;528(Pt 3):497-507. doi: 10.1111/j.1469-7793.2000.00497.x.
The perforated patch whole-cell configuration of the patch-clamp technique was applied to superficial cells in intact mouse pancreatic islets. Three types of electrical activity were observed corresponding to alpha-, beta- and delta-cells. The delta-cells were electrically active in the presence of glucose but lacked the oscillatory pattern seen in the beta-cells. By contrast, the alpha-cells were electrically silent at high glucose concentrations but action potentials could be elicited by removal of the sugar. Both alpha- and beta-cells contained transient voltage-activated K+ currents. In the delta-cells, the K+ currents activated above -20 mV and were completely blocked by TEA (20 mM). The alpha-cells differed from the delta-cells in possessing a TEA-resistant K+ current activating already at -40 mV. Immunocytochemistry revealed the presence of Kv3.4 channels in delta-cells and TEA-resistant Kv4.3 channels in alpha-cells. Thus the presence of a transient TEA-resistant current can be used to functionally separate the delta- and alpha-cells. A TTX-sensitive Na+ current developed in delta-cells during depolarisations beyond -30 mV and reached a peak amplitude of 350 pA. Steady-state inactivation of this current was half-maximal at -28 mV. The delta-cells were also equipped with a sustained Ca2+ current that activated above -30 mV and reached a peak of 60 pA when measured at 2.6 mM extracellular Ca2+. A tolbutamide-sensitive KATP channel conductance was observed in delta-cells exposed to glucose-free medium. Addition of tolbutamide (0.1 mM) depolarised the delta-cell and evoked electrical activity. We propose that the KATP channels in delta-cells serve the same function as in the beta-cell and couple an elevation of the blood glucose concentration to stimulation of hormone release.
采用膜片钳技术的穿孔膜片全细胞记录模式,对完整小鼠胰岛中的表层细胞进行研究。观察到三种电活动类型,分别对应α细胞、β细胞和δ细胞。δ细胞在葡萄糖存在时具有电活性,但缺乏β细胞中所见的振荡模式。相比之下,α细胞在高葡萄糖浓度下电活动静止,但去除葡萄糖后可诱发动作电位。α细胞和β细胞均含有瞬时电压激活的钾电流。在δ细胞中,钾电流在膜电位高于 -20 mV 时被激活,并被 20 mM 的四乙铵(TEA)完全阻断。α细胞与δ细胞的不同之处在于,其具有一种在 -40 mV 时就已激活的对 TEA 不敏感的钾电流。免疫细胞化学显示,δ细胞中存在 Kv3.4 通道,α细胞中存在对 TEA 不敏感的 Kv4.3 通道。因此,瞬时的对 TEA 不敏感电流的存在可用于从功能上区分δ细胞和α细胞。在δ细胞中,当去极化超过 -30 mV 时会产生一种对河豚毒素(TTX)敏感的钠电流,其峰值幅度达到 350 pA。该电流的稳态失活在 -28 mV 时达到半数最大。δ细胞还具有一种持续的钙电流,在膜电位高于 -30 mV 时被激活,当细胞外钙浓度为 2.6 mM 时测量,其峰值为 60 pA。在无糖培养基中培养的δ细胞中观察到一种对甲苯磺丁脲敏感的ATP敏感性钾通道电导。加入甲苯磺丁脲(0.1 mM)可使δ细胞去极化并诱发电活动。我们认为,δ细胞中的ATP敏感性钾通道与β细胞中的功能相同,将血糖浓度升高与激素释放刺激相偶联。