Neuroadaptations to Ethanol in Drosphila

描述（由申请人提供）：本研究项目的长期目标是阐明乙醇神经适应的分子和细胞机制。先前的研究已经确定了两个新的果蝇突变体，它们对乙醇的敏感性提高，耐受性降低。一个突变体的初步空间表达研究，命名为荷马，已经表明，正常基因的功能是足够的GABA能神经元，神经支配的椭圆体的正常行为反应，乙醇。这些研究将得到确认和扩展。此外，RNAi构建体和显性阴性的定向表达将用于定义成年脑中对乙醇的野生型行为反应所需的Corner表达的位置。新开发的技术，提供实验控制转基因表达的时间和空间将被用来确定是否homer是需要在发展过程中，或是否需要在成年苍蝇的生理。将使用第二个突变体的正常转基因（命名为yps）进行类似的时间和空间表达研究，以确定该基因的功能是否在同一组神经元中需要，并与正常乙醇反应的homer同时需要。来自需要homer和/或yps的神经元的RNA将使用新开发的亲和技术分离，并用于探针微阵列，以确定在这组特定神经元中乙醇暴露时发生的基因表达变化。 观察到的基因表达与乙醇暴露的变化将用于预测哪些基因在功能上参与乙醇的行为反应，这些基因的突变体将被测定，以进一步确定正常乙醇敏感性和耐受性的分子遗传要求。这些研究将进一步加深我们对酒精敏感性和耐受性的遗传基础的理解，这是酒精中毒易感性的重要参数。

DESCRIPTION (provided by applicant): The long-term goals of this research project are to elucidate the molecular and cellular mechanisms for neuroadaptations to ethanol. Previous studies have identified two new Drosophila mutants with a heightened sensitivity and impaired tolerance to ethanol. Preliminary spatial expression studies of one mutant, named homer, have suggested that the normal gene's function is sufficient in GABAergic neurons that innervate the ellipsoid body for normal behavioral responses to ethanol. These studies will be confirmed and extended. In addition, the directed expression of RNAi constructs and dominant negatives will be used to define where Corner expression is required in the adult brain for wild type behavioral responses to ethanol. Newly developed techniques that provide for experimenter control over transgene expression in time and space will be used to ascertain whether homer is required during development or whether it is required physiologically in the adult fly. Similar time and space expression studies using a normal transgene of the second mutant, named yps, will be performed to ascertain whether this gene's function is required in the same set of neurons and at the same time as homer for normal ethanol responses. RNA from neurons that require homer and/or yps will be isolated using newly developed affinity techniques and used to probe microarrays to define the changes in gene expression that occur with ethanol exposure in this specific set of neurons. The observed changes in gene expression with ethanol exposure will be used to predict which genes are functionally involved in behavioral responses to ethanol, and mutants in these genes will be assayed to further define the molecular genetic requirements for normal ethanol sensitivity and tolerance. These studies will further our understanding of the genetic basis for ethanol sensitivity and tolerance, important parameters for susceptibility to alcoholism.