Rapid functional evaluation of beta-cells by extracellular recording of membrane potential oscillations with microelectrode arrays
Rapid functional evaluation of beta-cells by extracellular recording of membrane potential oscillations with microelectrode arrays
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通过微电极阵列在细胞外记录膜电位振荡来快速评估 β 细胞的功能
DOI：
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
P. Krippeit
中科院分区：
文献类型：
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作者：
T. Pfeiffer;U. Kraushaar;M. Düfer;S. Schönecker;D. Haspel;E. Günther;G. Drews;P. Krippeit
当葡萄糖浓度在~6 ~ 25mm之间时，β细胞的膜电位（Vm）振荡，即爆发期与动作电位交替与沉默的爆发间期交替产生所谓的慢波。慢波驱动胞浆Ca2+浓度（[Ca2+]c）和胰岛素分泌的振荡。爆发的时间长短与胰岛素的释放量有关。因此，平台期（FOPP）的比例，即爆发活动的时间百分比，是β细胞功能和代谢完整性的极好标记。使用微电极阵列（MEA）测量小鼠胰岛细胞外电压变化，允许检测爆发期和爆发间期。在无刺激的葡萄糖浓度（3mm）下，没有检测到电活动，而在30mm时连续破裂。葡萄糖浓度-响应（以FOPP确定）曲线显示在12±1mm时半最大刺激（Hill方程拟合）。该信号对KATP通道调节剂敏感，如甲苯丁胺或二氮氧化物。同时记录的电活动和[Ca2+]c分别显示一致的爆发和峰值。细胞外记录与更耗时的细胞内电记录完全一致。该结果为在基于mea的系统中使用细胞外电极检测β细胞慢波和测定FOPP提供了“原理证明”。该方法简便，能够实时研究β细胞功能调节剂的作用，包括可能的抗糖尿病药物。此外，该方法可能有助于在移植前检查人类供体胰岛的代谢完整性。
The membrane potential (Vm) of beta-cells oscillates at glucose concentrations between ~6 and 25 mM, i.e. burst phases with action potentials alternate with silent interburst phases generating so-called slow waves. The slow waves drive oscillations of the cytosolic Ca2+ concentration ([Ca2+]c) and insulin secretion. The length of the bursts correlates with the amount of insulin release. Thus, the fraction of plateau phase (FOPP), i.e. the percentage of time with burst activity, is an excellent marker for beta-cell function and metabolic integrity. Extracellular voltage changes of mouse islets were measured using a microelectrode array (MEA) allowing the detection of burst and interburst phases. At a non-stimulating glucose concentration (3 mM) no electrical activity was detectable while bursting was continuous at 30 mM. The glucose concentration–response (determined as FOPP) curve revealed half-maximal stimulation at 12 ± 1 mM (Hill equation fit). The signal was sensitive to KATP channel modulators, e.g. tolbutamide or diazoxide. Simultaneous recordings of electrical activity and [Ca2+]c revealed congruent bursts and peaks, respectively. The extracellular recordings are in perfect agreement with more time-consuming intracellular electrical recordings. The results provide a 'proof-of-principle' for detection of beta-cell slow waves and determination of the FOPP using extracellular electrodes in a MEA-based system. The method is facile and provides the capability to study the effects of modulators of beta-cell function including possible anti-diabetic drugs in real time. Moreover, the method may be useful for checking the metabolic integrity of human donor islets prior to transplantation.
影响因子：
7.4
作者：
Dishinger, John F.;Reid, Kendra R.;Kennedy, Robert T.
通讯作者：
Kennedy, Robert T.
影响因子：
7.7
作者：
Khan, FA;Goforth, PB;Satin, LS
通讯作者：
Satin, LS