Genetic Analysis of Olfaction

描述（由申请人提供）：昆虫化学感受系统与脊椎动物化学感受系统有许多相似之处，但在数值上更简单。果蝇中可用的有效的分子遗传工具结合简单的化学感受行为测定和电生理分析，使我们能够确定化学感受系统的各个组成部分的体内生物学功能。最终，这些结果将提供深入了解化学感觉信息处理的基本原则，可能与脊椎动物共享。我们提出以下具体目标： 具体目标1。阐明感受器周围的事件和分子组成的敏感性，男性信息素，11-顺式醋酸乙烯酯。在上一个资助周期中，我们发现气味结合蛋白LUSH是化学感受神经元的一小部分中信息素敏感性所必需的。值得注意的是，LUSH似乎是嗅觉神经元的直接激活剂。我们将确定LUSH是否直接结合到11-cis vaccenyl acetate，并使用靶向RNAi方法和遗传方法鉴定由LUSH激活的神经元受体。我们将确切地建立信息素，LUSH和嗅觉神经元受体如何共同发挥作用来介导信息素信号。 具体目标2。将果蝇触角的功能性外周嗅觉图与解剖图相关联。我们将使用反向遗传学方法将特定气味受体基因产物的表达与嗅觉神经元的功能类别相关联。这将使我们能够合并的解剖外周嗅觉地图与功能地图，并建立在体内的气味特异性的每个气味受体中表达的天线。这些信息最终将有助于理解大脑如何将气味混合物转化为有意义的气味图像。我们将使用一个转基因RNAi方法首先描述了我的实验室的目标表达的个人气味受体和识别缺陷的功能类的转基因果蝇使用单感器电生理记录。 具体目标3。使用反向遗传方法来阐明在触角中表达的其他OBP蛋白的作用。我们已经确定了35个基因编码OBP蛋白在果蝇基因组中。16种气味结合蛋白在触角中表达。所有这些蛋白质都介导信息素反应，还是有些蛋白质参与了一般气味的检测？我们将使用反向遗传学来消除这些蛋白质在触角中的表达，并确定嗅觉神经元的功能类别或交配或聚集行为是否存在缺陷。

DESCRIPTION (provided by applicant): The insect chemosensory system has many parallels with vertebrate chemosensory systems, but is numerically simpler. The potent molecular genetic tools available in Drosophila combined with simple chemosensory behavior assays and electrophysiological analysis allow us to determine the in vivo biological function of individual components of the chemosensory system. Ultimately these results will provide insights into underlying principles of chemosensory information processing that are likely to be shared with vertebrates. We propose the following specific aims: Specific Aim 1. Elucidate the perireceptor events and molecular components underlying sensitivity to the male pheromone, 11-cis vaccenvl acetate. In the last funding cycle we discovered the odorant binding protein LUSH is required for pheromone sensitivity in a small subset of chemosensory neurons. Remarkably, LUSH appears to function as a direct activator of olfactory neurons. We will determine whether LUSH binds directly to 11-cis vaccenyl acetate, and identify the neuronal receptor that is activated by LUSH using a targeted RNAi approach and a genetic approach. We will establish exactly how the pheromone, LUSH and the olfactory neuron receptor function together to mediate pheromone signaling. Specific Aim 2. Correlate the functional peripheral olfactory map to the anatomic map in the Drosophila antenna. We will correlate expression of specific odorant receptor gene products with functional classes of olfactory neurons using a reverse genetic approach. This will allow us to merge the anatomic peripheral olfactory map with the functional map and establish the in vivo odorant specificity for each odorant receptor expressed in the antenna. This information will ultimately help understand how the brain converts odorant blends into meaningful odor images. We will use a transgenic RNAi approach first described by my laboratory to target expression of individual odorant receptors and identify defective functional classes in the transgenic flies using single-sensillum electrophysiological recordings. Specific Aim 3. Use a Reverse Genetic Approach to elucidate the role of the other OBP proteins expressed in the antenna. We have identified 35 genes encoding OBP proteins in the Drosophila genome. 16 odorant binding proteins are expressed in the antenna. Are all of these proteins mediating pheromone responses or are some involved in detection of general odorants? We will use reverse genetics to eliminate expression of these proteins in the antenna and determine if there are defects in functional classes of olfactory neurons or in mating or aggregation behavior.