MECHANISMS OF CELL MEMBRANE SIGNALS

跨膜信号传导机制，特别是非兴奋性 脊椎动物细胞对外界刺激的电磁反应， 被研究。 单分子通道电导的原位记录 细胞膜以及重组到脂质体中， 发现和测量已知和预期信道的属性。 微量移液器千兆欧姆密封件将用于整个囊泡记录， 直接测量由重组泵产生的电流。 具有数字图像分析的电光图像增强视频显微镜 将与荧光指示剂一起使用， 非兴奋细胞的反应，以测量空间分布 膜电位和表面受体上的配体，并检测信号 与表面受体交联相关，应用中的电迁移 电化学领域和内吞过程。 技术 旨在增强荧光显微照相术的能力， 研究以前无法获得的跨膜性质 信令将被追踪。 荧光膜电位pH和钙 离子指示染料将用于空间分布的绘图 细胞膜电位和离子活性的变化。 大 将操纵带有重构蛋白质通道的脂质囊泡 为了开发适合于全细胞片记录的形态， 以及我们目前可用的单通道录音。 新 战略，利用我们的同时敏感的光学和 将探索细胞生物学中的电记录能力。

Mechanisms of transmembrane signaling, particularly non-excitable electromagnetic responses of vertebrate cells to external stimuli, are to be studied. Recording of single molecular channel conductances in situ in cell membranes and also as reconstituted into liposomes is to be used to discover and to measure properties of known and anticipated channels. Micropipette gigaohm seals are to be used for whole vesicle recording for direct measurements of currents generated by reconstituted pumps. Electro-optical image enhanced video microscopy with digital image analysis are to be used with flourescent indicators to search for electrogenic responses in nonexcitable cells, to measure spatial distributions of membrane potentials and ligands on surface receptors, and to detect signals associated with surface receptor cross linking, electromigration in applied electrochemical fields and in endocytotic processes. Technical developments designed to enhance the power of fluorescence micrography for investigation of previously inaccessible properties of transmembrane signaling will be pursued. Fluorescent membrane potential pH and calcium ion indicating dyes will be applied to mapping of the spatial distribution of membrane potentials and ion activities in differentiated cells. Large lipid vesicles bearing reconstituted protein channels will be manipulated in order to develop morphologies suitable for whole cell patch recording, as well as our presently available single channel recording. New strategies to take advantage of our simultaneous sensitive optical and electrical recording capability in cell biology will be explored.