Marszalek P E, Markin V S, Tanaka T, Kawaguchi H, Fernandez J M
Department of Physiology and Biophysics, Mayo Clinic, Rochester, MN 55905, USA.
Biophys J. 1995 Oct;69(4):1218-29. doi: 10.1016/S0006-3495(95)80004-4.
When placed at the tip of a glass micropipette electrode the polymeric matrix of the secretory granule behaves like a diode. The measured current was 100-fold greater at negative potentials compared to positive potentials, and up to sixfold greater than that measured with the pipette alone. By manipulating the geometry of the electric field we show that these electrical properties result from focusing an electric field at the gel-electrolyte interface. We also show, by using pulsed-laser imaging with fluorescein as the ionic probe, that there is a rapid accumulation and depletion of ions at the gel-electrolyte interface. A voltage pulse of -9 V applied to the gel caused a severalfold increase in the fluorescence intensity within 5 ms. This correlated with an increase in the measured current (approximately 1 microA). In contrast, within 5 ms of applying +9 V we recorded a decrease in the fluorescence intensity, which paralleled the twofold decrease in the measured current. This is similar to a p-n junction where an applied voltage causes the accumulation and depletion of charge carriers. Using synthetic gels (diameter 3-6 microns) with different charge characteristics we observed no rectification of the current with neutral gels and confirmed that rectification and amplification of the current were dependent on the fixed charge within a gel. In addition, we modeled the conduction at the gel-electrolyte interface using the Nernst-Planck electrodiffusion equation and accurately fitted the experimental current-voltage relationships. This study provides some insight into how biological interfaces may function. For example, we suggest that neurotransmitter release during exocytosis could be regulated by voltage-induced accumulation and depletion of ions at the interface between the secretory granule and the fusion pore.
当置于玻璃微吸管电极尖端时,分泌颗粒的聚合物基质表现得像一个二极管。与正电位相比,负电位下测得的电流大100倍,比单独用吸管测得的电流大6倍。通过操纵电场的几何形状,我们表明这些电学性质是由电场在凝胶 - 电解质界面处的聚焦产生的。我们还通过使用以荧光素为离子探针的脉冲激光成像表明,在凝胶 - 电解质界面处存在离子的快速积累和消耗。施加到凝胶上的 -9 V电压脉冲在5毫秒内使荧光强度增加了几倍。这与测得的电流增加(约1微安)相关。相反,在施加 +9 V的5毫秒内,我们记录到荧光强度下降,这与测得的电流下降两倍平行。这类似于p-n结,其中施加的电压会导致电荷载流子的积累和消耗。使用具有不同电荷特性的合成凝胶(直径3 - 6微米),我们观察到中性凝胶没有电流整流现象,并证实电流的整流和放大取决于凝胶内的固定电荷。此外,我们使用能斯特 - 普朗克电扩散方程对凝胶 - 电解质界面处的传导进行建模,并准确拟合了实验电流 - 电压关系。这项研究为生物界面如何发挥作用提供了一些见解。例如,我们认为胞吐过程中神经递质的释放可能受电压诱导的离子在分泌颗粒与融合孔之间界面处的积累和消耗调节。