Microanalytical Methods for Drosophila Neurochemistry

描述（由申请人提供）：本研究的总体目标是开发新的微分析方法来研究果蝇的神经化学。果蝇可以说是生物学中研究最多的生物体之一，作为模型系统具有许多优势。然而，需要少量且灵敏的分析方法来检查与果蝇遗传和行为方面相关的化学变化，这是一个困难。果蝇神经系统包含大约 200,000 个细胞，仅占据约 8 nL。我们建议进一步开发和应用我们最近报道的针对果蝇研究的小样品处理和毛细管电泳方法。这些方法已经产生了一些迄今为止报道的用于生物样品电检测的最丰富的电泳图或色谱图，并且有许多我们尚未鉴定的化合物被洗脱。此外，我们建议利用 Pi 在体内伏安法中的背景来开发果蝇体内伏安法的方案。这将提供完整有机体中最小的体内方法，并且是一个令人兴奋的分析挑战。当前和新开发的方法将应用于慢性酒精耐受性的化学机制和单只苍蝇异常值的分析。该提案的目的是 1) 增强用于分析分离果蝇匀浆、单头和神经组织切片的采样方法，2) 开发体内电化学方法，用于对不同果蝇大脑结构的动力学进行化学分析，3) 应用新开发的技术来识别与慢性酒精耐受相关的神经化学物质。

DESCRIPTION (Provided by applicant): The overall objective of this research is to develop new microanalytical approaches to study the neurochemistry of Drosophila melanogaster, the fruit fly. Drosophila is arguably one of the most studied organisms in biology offering many advantages as a model system. However, there is a difficulty with a need for low-volume and sensitive analytical methods to examine the chemical changes associated with genetic and behavioral aspects of the fly. The fly nervous system contains approximately 200,000 cells and occupies only about 8 nL. We propose to further develop and apply small sample handling and capillary electrophoresis methods we have recently reported for studies with the fly. These methods have produced some of the richest electropherograms or chromatograms for electrical detection of a biological sample reported to date and there are many compounds eluted that we have not yet identified. In addition, we propose to use one Pi's background in in vivo voltammetry to develop protocols for in vivo voltammetry in the fly. This will present the smallest in vivo approach in an intact organism and is an exciting analytical challenge. Current and newly developed methods will be brought to bear on the chemical mechanisms that underlie chronic alcohol tolerance and the analysis of single fly outliers. The aims of the proposal are to 1) enhance sampling methodologies for analytical separations of fly homogenates, single heads, and sections of nervous tissue, 2) develop in vivo electrochemical methods for the chemical analysis of dynamics in distinct fly brain structures, and 3) apply the newly developed techniques to identify neurochemicals associated with chronic tolerance to alcohol.