IDENTIFICATION OF ACOUSTICO-LATERALIS TRANSMITTERS

研究的目的是从化学上确定 豚鼠耳蜗核的外周递质， 非洲爪蟾（非洲爪蟾）的侧线器官， 鱼的消化器官，并表征生化系统 与假定发射机相关联。 分析方法 包括：1）高分辨率、高效液相色谱（HPLC） 用荧光检测和放射性检测，2）质量 光谱法，用于确认HPLC级分的化学组成， 3)放射性配体-受体结合测定和放射自显影定位， 用于表征听外侧神经递质受体 馏分，4）荧光和放射性同位素测定，用于测定 毛细胞组分的传递相关酶，和5）使用 非洲爪蟾侧线，用于检测类似发射器的活动。 治疗方法包括：1）手术入路 耳蜗和颞骨，用于收集外淋巴液和脑脊液 在存在和不存在噪声的情况下， 测定流体的小分子含量，2）用于 从听-外侧组织释放假定的神经递质， （3）爪蟾侧线、鱼体的解剖， 豚鼠听觉分数，在确定其 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，并在 侧线神经肥大和鱼类神经系统器官，推测 声-侧-是发射机（S），其（它们）相关的受体， 合成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， 和假定的豚鼠耳蜗递质的生物活性。 我们进一步计划确定化学上非发射器的材料， 豚鼠暴露于高强度噪声时外淋巴液中淋巴细胞的变化 程度. 这种主要的生物化学方法应该导致 识别听觉和平衡的外周神经递质， 并将为与发射机相关的听力损失提出最终的治疗方法， 耳鸣头晕

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.