MODULATION OF CA CURRENT IN APLYSIA BAG CELL NEURONS

海兔袋细胞神经元中 CA 电流的调节

• 批准号: 3084950
• 负责人: ELIZABETH JONAS
• 金额: $ 8.11万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-08-01 至 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3084950
• 关键词: Gastropoda; calcium flux; fluorescent dye /probe; neural transmission; neurons; neuropeptides; protein kinase C; second messengers; secretion; tissue /cell culture; voltage /patch clamp

As part of the training program of the applicant, this research project's aim is to elucidate further the function of calcium in neuromodulation of ion currents and in secretion, and to study the effects of neuroactive peptides on the calcium currents in Aplysia bag cell neurons. By using whole cell voltage clamping and single channel recording techniques, and by looking at the calcium concentration using fura-2 and image processing techniques, the changes in calcium current and intracellular calcium concentration in response to bag cell peptides can be studied in the intact nervous system. A set of experiments will attempt to determine if the peptides work by activating protein kinase C. The effects of the peptides on the calcium currents will be examined in the presence of protein kinase inhibiting agents to see if observed changes in calcium current can be prevented in the presence of these substances. The effect of the peptide on calcium current will be examined during the neurons' refractory period to determine if calcium and neuroactive peptides play a role in bringing on or maintaining the refractory period. Bag cell neurons will be examined in primary culture to study specific properties of the ion currents and the influence of the peptides on the currents. The influence of culturing the neurons on the calcium current will be studied. In order to see if ion channels can move during the life of a cell, cultured bag cell neurons will be examined using electrophysiological techniques and fluorescence imaging techniques to look at changes in the concentration of calcium in the soma as compared to the neurites during the process of neurite-sprouting. Cytoskeleton inhibiting agents will used to try to inhibit changes in calcium current observed. Neurons will be studied in the absence of external calcium to see if autoactive peptides can influence the release of intracellular calcium. The ultimate goal of this research will be to determine how neurons communicate in order to provide insight into prevalent neuropathological processes.

作为申请人培训计划的一部分，本研究项目 目的是进一步阐明钙在神经调节中的作用， 离子电流和分泌，并研究神经活性的影响 肽对豚鼠囊细胞钙电流的影响。通过使用 全电池电压箝位和单通道记录技术， 用Fura-2和图像处理技术观察钙浓度 技术，钙电流和细胞内钙的变化 响应于袋细胞肽的浓度可以在完整的细胞中研究。 神经系统一组实验将试图确定 肽通过激活蛋白激酶C起作用。肽的作用 将在蛋白激酶存在下检查对钙电流的影响 抑制剂，以观察观察钙电流的变化是否可以 在这些物质存在的情况下。所述肽对以下的影响： 将在神经元的不应期检查钙电流， 确定钙和神经活性肽是否在引起或 维持不应期。袋细胞神经元将在 原代培养，以研究离子电流的特定特性， 肽对电流的影响。文化的影响 将研究神经元对钙电流的影响。为了观察离子 通道可以在细胞的生命过程中移动，培养的袋细胞神经元将 使用电生理技术和荧光成像进行检查 观察索马中钙浓度变化的技术 与神经突出芽过程中的神经突相比。 细胞骨架抑制剂将用于试图抑制细胞骨架的变化。 观察到钙电流。神经元将在缺乏 外部钙，看看自身活性肽是否可以影响释放 胞内钙这项研究的最终目标是 确定神经元是如何交流的， 普遍的神经病理过程。