MECHANISMS OF ADAPTATION AND EXCITATION

适应和兴奋机制

• 批准号: 3483901
• 负责人: JOHN E LISMAN
• 金额: $ 26.74万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-06-01 至 1992-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3483901
• 关键词: Anura; Xenopus; antibody; calcium; cyclic nucleoside monophosphate; cytoplasm; dark adaptation; dialysis; electrical potential; electrophysiology; fresh water environment; horseshoe crabs; membrane channels; neurotransmitters; photochemistry; rod cell; saltwater environment; visual photoreceptor; visual photosensitivity; visual stimulus

The general goal of this proposal is elucidation of the mechanism by which light generates the receptor potential in photoreceptors. This process can be divided into early stages involving the light- sensitive molecule rhodopsin, intermediate stages involving chemical amplification, and the final stage in which the chemical message affects ion channels, thereby generating the receptor potential. An aspect of the early steps, the average number of G- proteins activated by a single isomerized rhodopsin, will be investigated in experiments involving agents which block or stimulate G-protein. In addition the possibility of using peptides to inhibit specific reactions involving rhodopsin will be explored by dialyzing such peptides into tiger salamander rods or Limulus photoreceptors. The main questions involving intermediate events have to do with the new hypothesis that a rise in cGMP mediates the excitation of invertebrate photoreceptors. This will be studied in biochemical assays of the effect of light on guanylate cyclase activity, phosphodiesterase activity, and the concentration of cGMP in the living squid eye. Physiological methods will be used to study the effect of putative activators and inhibitors of enzymes involved in cyclic nucleotide metabolism. The general goal is to determine which enzyme is responsible for light-dependent changes in cGMP and how this enzyme might be controlled. Finally, anatomical methods will be used to identify the structural basis for the very inhomogeneous sensitivity to injected cGMP. Ongoing work on the light- activated channels in Limulus using the patch clam method will be continued. Preliminary results indicate that these channel events have two sizes. We will attempt to determine if these are due to two different states of the same channel or to two different channel types. These channels will also be studied in excised patches where it is possible to test whether substances of interest can have direct effects on the channel.

这项建议的总体目标是阐明这一机制 光通过它产生光感受器的感受器电势。 这个过程可以分为早期阶段，涉及光- 敏感分子视紫红质，中间阶段涉及 化学放大，以及化学放大的最后阶段 信息影响离子通道，从而产生受体 潜力。早期步骤的一个方面是，G-R的平均数量 由单一异构化视紫红质激活的蛋白质，将是 在涉及阻滞剂的实验中进行研究 刺激G蛋白。此外，使用多肽的可能性 为了抑制涉及视紫红质的特定反应，将进行探索 通过将这些多肽透析成老虎火蜥蜴棒或贻贝 光感受器。涉及中间事件的主要问题 与cGMP升高在 无脊椎动物光感受器的兴奋。这将是 光对鸟粪酸盐影响的生化分析研究 循环酶活性、磷酸二酯酶活性和 活体鱿鱼眼中cGMP的浓度。生理学 将使用各种方法来研究假定的激活剂的效果 与环核苷酸有关的酶的抑制剂 新陈代谢。总的目标是确定哪种酶是 负责cGMP的光依赖变化以及这是如何 酶可能会受到控制。最后，解剖学方法将是 用来确定非常不均匀的结构基础 对注射cGMP敏感。正在进行的关于光的工作- 使用补丁文蛤方法激活的通道将是 继续。初步结果表明，这些渠道事件 有两个尺码。我们将尝试确定这些是否是由于 同一通道的两个不同状态或两个不同的状态 频道类型。这些通道也将在实验中进行研究 可以测试感兴趣物质的补丁 可以对频道产生直接影响。