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之一，作为一个模型系统提供了许多优势。然而，需要小容量和敏感的分析方法来检查与苍蝇的遗传和行为方面相关的化学变化是一个困难。苍蝇的神经系统包含大约20万个细胞，只占大约8立方千米。我们建议进一步发展和应用我们最近报道的小样本处理和毛细管电泳方法用于苍蝇的研究。这些方法已经产生了一些最丰富的电检测生物样品的电泳图或色谱图，有许多化合物洗脱，我们还没有确定。此外，我们建议使用一个Pi的背景在体内伏安法来制定在苍蝇体内伏安法的方案。这将在一个完整的有机体中呈现最小的体内方法，这是一个令人兴奋的分析挑战。现有的和新开发的方法将用于研究慢性酒精耐受性的化学机制，并分析单个苍蝇的异常值。该提案的目的是：1)加强苍蝇匀浆、单个头部和神经组织切片分析分离的采样方法；2)发展体内电化学方法，用于不同苍蝇大脑结构动力学的化学分析；3)应用新开发的技术来识别与慢性酒精耐受性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.