VOLATILE ANESTHETICS AND NEURONAL CALCIUM

挥发性麻醉剂和神经钙

• 批准号: 6519571
• 负责人: THOMAS Joseph John BLANCK
• 金额: $ 13.47万
• 依托单位: HOSPITAL FOR SPECIAL SURGERY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 2002-12-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6519571
• 关键词: G protein; calcium channel; calcium flux; carbachol; chemical binding; electrostimulus; halothane; hippocampus; homeostasis; inhalation anesthesia; isoflurane; laboratory rat; neurons; neuropharmacology; norepinephrine; tissue /cell culture; voltage gated channel

Neurons translate electrical and chemical stimuli into physiologically meaningful information by Ca2+ translocation and redistribution. The hypothesis the investigators will test in the proposed studies is that the volatile anesthetics (VA), halothane and isoflurane, interfere with normal extra- and intracellular Ca2+ translocation and distribution, and that the VA effect on Ca2+ homeostasis (translocation and redistribution) depends on the type of stimuli. Studies will be performed in a human neuroblastoma cell line, SH-SY5Y cells, that has many characteristics of adrenergic neurons, in rat hippocampal neurons in primary culture, and in HEK293 cells transfected with alpha and beta subunits of N-type Ca2+ channels. Using intracellular Ca2+ fluorescence, single channel and whole cell patch-clamp, radioisotopic measurement of noradrenaline (NA) secretion, and molecular biologic techniques to generate N-type Ca2+ channels, we will accomplish the following specific aims: 1) identify the mechanisms mediating the VA-induced increase in the K+-evoked [Ca2+]cyt transient and determine if VA also enhance [Ca2+]cyt transients resulting from electrical stimulation; 2) identify the mechanisms that lead to the VA-induced depression of the Carbachol-evoked [Ca2+]cyt; transient; 3) investigate the source of Ca2+ that modulates the VA action on neuronal L-type Ca2+ channels; 4) investigate the VA action on N-type Ca2+ channels, and the involvement of G-proteins; and 5) investigate which sources of Ca2+ are important for mediating NA release, whether VA exposure alters the relationship between [Ca2+]cyt and NA release, and determine whether the VA action on NA release is dependent on the exposure time to Vas. The results of these investigations will help to establish that modifications of Ca2+ homeostasis are a fundamental effect of the Vas and that they are directly related to the process of anesthesia. This study will lead to the identification of organelles and molecules that are targets of VA's that will assist in the future development of more specific anesthetic drugs.

神经元将电和化学刺激转化为 CA2+易位和 重新分布。调查人员将在提议的假设中测试 研究是挥发性麻醉药（VA），氟烷和异氟烷， 干扰正常的细胞外Ca2+易位和 分布，以及VA对Ca2+稳态的影响（易位和 重新分布）取决于刺激的类型。研究将在 人类神经母细胞瘤细胞系，SH-SY5Y细胞，具有许多特征 肾上腺能神经元，在原发性培养中的大鼠海马神经元中 HEK293细胞用N型Ca2+通道的α和β亚基转染。 使用细胞内Ca2+荧光，单通道和全细胞 斑夹，去甲肾上腺素（NA）分泌的放射性分析测量和 分子生物学技术生成N型Ca2+通道，我们将 完成以下特定目的：1）确定介导的机制 VA引起的K+诱发[Ca2+] Cyt瞬态的增加，并确定是否是否 VA还增强了由电刺激导致的[Ca2+] Cyt瞬变； 2） 确定导致VA引起的抑郁的机制 卡尔巴乔诱发的[Ca2+] Cyt;瞬态； 3）调查Ca2+的来源 调节对神经元L型Ca2+通道的VA作用； 4）调查VA 对N型Ca2+通道的作用以及G蛋白的参与； 5） 研究哪些Ca2+来源对于中介Na释放很重要， VA暴露是否改变[Ca2+] Cyt和Na释放之间的关系， 并确定NA释放的VA作用是否取决于暴露 是时候到达Vas。这些调查的结果将有助于确定 CA2+稳态的修改是VA的基本效果，并且 它们与麻醉过程直接相关。这项研究将导致 识别是VA目标的细胞器和分子 将有助于未来开发更具体的麻醉药。