Neuroadaptations to Ethanol in Drosphila

描述(申请人提供)：本研究项目的长期目标是阐明神经适应酒精的分子和细胞机制。之前的研究已经发现了两个新的果蝇突变体，它们对乙醇的敏感度提高，耐受性减弱。一种名为HOMER的突变体的初步空间表达研究表明，正常基因的功能在支配椭球体的GABA能神经元中足以对乙醇做出正常的行为反应。这些研究将得到证实和推广。此外，RNAi结构和显性负值的定向表达将被用来定义成人大脑中对乙醇的野生型行为反应需要Corner的表达。新开发的技术允许实验者在时间和空间上控制转基因表达，将用于确定在发育过程中是否需要荷马，或者成年苍蝇是否需要生理上的荷马。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发挥作用，并同时作为正常乙醇反应的荷马。需要HOMER和/或YP的神经元的RNA将使用新开发的亲和技术分离出来，并用于探针微阵列，以确定酒精暴露后这组特定神经元中基因表达的变化。 观察到的酒精暴露后基因表达的变化将被用来预测哪些基因在功能上参与了对酒精的行为反应，并将对这些基因的突变进行分析，以进一步确定正常酒精敏感性和耐受性的分子遗传学要求。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.