Neuroadaptations to Ethanol in Drosophila

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种新的果蝇突变体，它们对乙醇的敏感性提高，耐受性受损。一种名为homer的突变体的初步空间表达研究表明，正常基因的功能足以控制椭球体的gaba能神经元对乙醇的正常行为反应。这些研究将得到证实和推广。此外，RNAi结构的定向表达和显性阴性将用于确定野生型乙醇行为反应中成人大脑中需要Corner表达的位置。新开发的技术提供了实验者控制转基因在时间和空间上的表达，将用于确定在发育过程中是否需要荷马，或者在成年果蝇中是否需要荷马。将使用第二个突变体的正常转基因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需要在同一组神经元中，并同时用于正常的乙醇反应。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进行分离，并用于探针微阵列，以确定乙醇暴露在这组特定神经元中发生的基因表达变化。

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.