IDENTIFICATION OF ACOUSTICO-LATERALIS TRANSMITTERS

这项研究的目的是在化学上确定假定的 豚鼠耳蜗核的外周递质(S) 非洲爪蛙(非洲爪蛙)的侧线器官 鱼的迷宫器官，并对生化系统进行了表征 与推定发送者(S)有关。分析方法 包括：1)高分辨率、高性能的高效液相色谱仪 用荧光检测和放射性检测，2)质量 光谱分析，用于确认高效液相色谱组分的化学成分， 3)放射配基-受体结合分析和放射自显影定位； 用于表征听觉外侧肌的神经递质受体 4)荧光和放射性同位素分析，用于测定 毛细胞组分的递质相关酶，以及5)使用 非洲爪哇的侧线，用于检测类传送器的活动。 准备方法包括：1)对豚鼠进行手术入路 耳蜗骨和颞骨，用于采集外淋巴液和脑脊液 豚鼠在存在和不存在噪音的情况下产生的液体 测定流体的小分子含量，2)体外系统 从听觉侧方组织释放假定的神经递质， (3)非洲爪蛙侧线、鱼迷路的解剖； 豚鼠听觉部分，在测定其 小分子含量、酶活性和受体含量。使用这些 方法，我们计划建立身份，并存在于 推定的横线神经肥大和鱼的迷路器官 声-侧-IS递质(S)及其相关受体和 合成酶和降解酶及其生物活性。 我们还计划建立身份，刺激释放到外淋巴， 和推测的豚鼠耳蜗神经递质的生物活性(S)。 我们进一步计划确定化学上非传递性材料是 豚鼠暴露于高强度噪声时释放到外淋巴中 级别。这种以生化为主的方法应该会导致 听力与平衡外周神经递质(S)的鉴定 并将建议最终治疗与发射器相关的听力损失的方法， 耳鸣和头晕。

The objective of the research is to identify chemically the presumptive peripheral transmitter(s) of the cochlear nucleus of the guinea pig, the lateral-line organ of Xenopus laevis (the African clawed frog), and of labyrinthine organs of fish, and to characterize the biochemical systems associated with the presumptive transmitter(s). Analytical methods include: 1) high-resolution, high-performance liquid chromatography (HPLC) with fluorescence detection and detection by radioactivity, 2) mass spectrometry, for confirmation of chemical composition of HPLC fractions, 3) radioligand-receptor binding assays and autoradiographic localization, for characterization of neurotransmitter receptors of acoustico-lateralis fractions, 4) fluorometric and radioisotopic assays, for determination of transmitter-related enzymes of hair-cell fractions, and 5) a bioassay using the Xenopus lateral line, for detecting transmitter-like activity. Preparative methods include: 1) a surgical approach to the guinea-pig cochlea and temporal bone, for collection of perilymph and cerebrospinal fluid from guinea pigs in the presence and absence of noise, before determining small-molecule content of the fluids, 2) an in vitro system for release of presumptive neurotransmitters from acoustico-lateralis tissues, and 3) dissection of Xenopus lateral line, fish labyrinthine, and guinea-pig auditory fractions, prior to determination of their small-molecule content, enzyme activity, and receptor content. Using these methods, we plan to establish the identity, and presence in the lateral-line neuromast and fish labyrinthine organs, of presumptive acoustico-lateral-is transmitter(s), its (their) associated receptors and synthesizing and degrading enzymes, and its (their) biological activity. We also plan to establish the identity, stimulated release into perilymph, and biological activity of presumptive guinea-pig cochlear transmitter(s). We further plan to identify chemically non-transmitter materials that are releasted into perilymph during exposure of guinea pigs to noise at high levels. This predominantly biochemical approach should lead to the identification of peripheral neurotransmitter(s) of hearing and balance, and will suggest eventual therapies for transmitter-related hearing loss, tinnitus, and dizziness.