IDENTIFICATION OF ACOUSTICO-LATERALIS TRANSMITTERS

描述(摘自申请人的摘要)。该计划的主要目标是 目前的建议是识别外围设备 神经递质/神经调节剂及相关的生化系统 毛细胞器官。外周神经递质电压门控钙离子 通道和神经递质受体将被检测为哺乳动物 和鱼类模型系统。主要的假说解决了存在和 非谷氨酸毛细胞递质功能(S)，非L型电压- 门控钙通道(S)与新近报道的传出神经递质 乙酰胆碱和多巴胺的受体。方法包括：1)高性能 分辨率、高效液相色谱(HPLC)和检测 通过电化学、荧光、放射性和放射免疫分析，2) 组织含量分析及去极化诱导体外释药 球状毛发中推定的神经递质和神经调质 毛细胞是其唯一完整的细胞类型的单元格，以及 声诱导耳蜗外淋巴液的体内释放3)生物测定 利用非洲爪哇侧线的化合物，4)形态 分子实体的免疫化学定位和原位定位 杂交方法，5)分子功能的序列分析 电压门控逆转录-聚合酶链式反应 与毛细胞释放递质有关的钙通道，以及6) 传出神经递质RT-PCR后的功能序列分析 乙酰胆碱和多巴胺的受体。使用这些方法，它是 计划对其进行化学鉴定和生物活性测定 钙依赖的球囊感觉细胞释放化合物 通过低水平钾去极化并释放到外淋巴中 通过声音刺激。神经活性单胺及其相关物质的生物合成 分子将利用放射性前体和高效液相色谱进行研究。我们 将展示毛发的分子特征和定位 细胞相关的非L型电压门控钙通道和传出钙通道 相关的烟碱受体和多巴胺D2(ING)和D1受体。这 方法，利用微生物化学的方法，应该导致 继续详细阐述其结构和分子功能 听力和平衡的外周神经递质系统，指向 开发治疗与递质相关的听力损失的方法， 眩晕和耳鸣。

DESCRIPTION (from the Applicant's abstract). The major objective of the present proposal is to identify peripheral neurotransmitters/neuromodulators and associated biochemical systems of hair-cell organs. Peripheral neurotransmitters voltage-gated calcium channels, and neurotransmitter receptors will be examined for mammalian and fish model systems. The main hypotheses address the existence and function of non-glutamate hair cell transmitter(s), non-L-type voltage- gated calcium channel(s), and recently-described efferent neurotransmitter receptors for acetylcholine and dopamine. Methods include: 1) high- resolution, high-performance liquid chromatography (HPLC) with detection by electrochemistry, fluorescence, radioactivity, and radioimmunoassay, 2) analysis of tissue content, and depolarization-induced release in vitro of presumptive neurotransmitters and neuromodulators from a saccular hair cell sheet for which the hair cell is the only intact cell type, and sound-induced release in vivo into cochlear perilymph, 3) biological assay of compounds utilizing Xenopus laevis lateral line, 4) morphological localization of molecular entities by immunochemical and in situ hybridization methods, 5) sequence analysis as to molecular function after RT-PC (reverse transcription polymerase chain reaction) of voltage-gated calcium channels associated with transmitter release of haircells, and 6) functional sequence analysis after RT-PCR of efferent neurotransmitter receptors for acetylcholine and dopamine. Using these methods, it is planned to identify chemically and determine the biological activity of compounds release from saccular sensory cells in a calcium-dependent manner by low level potassium depolarization and released into perilymph by sound stimulation. Biosynthesis of neuroactive monoamines and related molecules will be studied utilizing radioactive precursors and HPLC. We will demonstrate molecular characteristics and localization of a hair cell-associated, non-L-type voltage-gated calcium channel and efferent- related a. nicotinic receptor and dopamine D2(ing) and D, receptors. This approach, utilizing methods of micro-biochemistry, should result in continued, detailed elucidation of structure and molecular function of peripheral neurotransmitter systems of hearing and balance, pointing the way to development of therapies for transmitter-related hearing loss, vertigo, and tinnitus.