MODULATION OF CA CURRENT IN APLYSIA BAG CELL NEURONS

作为申请者培训计划的一部分，本研究项目 目的是进一步阐明钙在神经调节中的作用。 离子电流和分泌物，并研究神经活性的影响 多肽对海兔袋状细胞神经元钙电流的影响。通过使用 全单元电压钳位和单通道记录技术，以及 用Fura-2和图像处理技术检测钙离子浓度 技术，钙电流和细胞内钙的变化 对BAG细胞多肽的反应浓度可以在完整的 神经系统。一系列实验将试图确定 多肽通过激活蛋白激酶C发挥作用。 在蛋白激酶存在的情况下，将检测钙电流 抑制剂观察观察到的钙电流变化是否可以 在这些物质存在的情况下被阻止的。多肽对血管内皮细胞生长的影响 在神经元的不应期将检测钙电流以 确定钙和神经活性多肽是否在发病中起作用 维持不应期。袋状细胞神经元将在 原代培养，以研究离子电流的特定性质和 多肽对电流的影响。养殖业的影响 将对神经元上的钙电流进行研究。为了看看离子是否 在细胞的生命过程中，通道可以移动，培养的袋状细胞神经元将 使用电生理技术和荧光成像进行检查 观察胞体内钙浓度变化的技术 与轴突萌发过程中的轴突进行比较。 细胞骨架抑制剂将被用来试图抑制 观察到钙电流。神经元的研究将在没有 外源性钙离子是否能影响自身活性多肽的释放 细胞内钙离子。这项研究的最终目标将是 确定神经元是如何通信的，以便深入了解 普遍的神经病理过程。

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.