Target Specificity in the Drosophila Olfactory System

描述(申请人提供)：果蝇的嗅觉系统为探索神经元在发育过程中形成联系的机制提供了一个独特的机会。表达相同气味受体的嗅觉感受器神经元(ON)连接到触角叶中的相同神经元，而表达不同气味受体的神经元连接到不同的神经元。这里提出的研究旨在了解这种特异性的分子基础。遗传学研究表明，不同的神经元使用不同的细胞表面蛋白簇来控制特异性，包括Dscam、N-钙粘蛋白、LAR受体酪氨酸磷酸酶和新的钙粘蛋白Flamingo。特别令人感兴趣的是对DSCAM的要求。DSCAM可以选择性地剪接，并有许多不同的形式(约38,000)，这可能有助于识别特异性。在这项提案中，将测试不同形式的DSCAM在控制嗅觉系统中神经元连接的特异性方面的作用。提出了三组实验，以批判性地评估替代形式的DSCAM在ORN靶向中的重要性。首先，已经分离出一组删除了许多不同形式的Dscam的删除。这些突变对一组三种不同的Orn的影响将在这些突变体中进行评估。其他缺乏其他形式的突变体将被分离，并将探索它们对ORN靶向的影响。其次，利用同源重组将含有单一胞外结构域的Dscam引入内源Dscam基因座。这将把多样性从大约38,000种减少到2种。将对三组嗅觉神经元的表型进行评估。最后，单一形式的Dscam将在野生型和突变型背景下的异源启动子下表达。将评估由这些形式诱导的主要表型。不诱导显性表型的形式将被评估为挽救完全丧失功能的Dscam突变的能力。更好地了解神经元连接是如何在发育过程中形成的，将有助于深入了解在各种神经和精神疾病中可能被破坏的机制。

DESCRIPTION (provided by applicant): The olfactory system in Drosophila melanogaster provides a unique opportunity to explore the mechanisms by which neurons form connections during development. Olfactory receptor neurons (ORNs) that express the same odorant receptors connect to the same neurons in the antennal lobe, while neurons that express different odorant receptors connect to different neurons. The studies proposed here are directed towards understanding the molecular basis of this specificity. Genetic studies have revealed that different neurons use different constellations of cell surface proteins to control specificity including Dscam, N-cadherin, Lar receptor tyrosine phosphatase and the novel cadherin, Flamingo. Of particular interest is the requirement for Dscam. Dscam is alternatively spliced and comes in many different forms (approximately 38,000) that may contribute to recognition specificity. In this proposal, the role of different forms of Dscam in controlling the specificity of neuronal connections in the olfactory system will be tested. Three sets of experiments are proposed to critically assess the importance of alternative forms of Dscam in ORN targeting. First, a set of deletions has been isolated that remove many different forms of Dscam. The effect of these mutations on a set of three different ORNs will be assessed in these mutants. Additional mutants lacking other forms will be isolated and their affects on ORN targeting will be explored. Second, homologous recombination will be used to introduce a single extracellular domain containing form of Dscam into the endogenous Dscam locus. This will reduce diversity from approximately 38,000 to 2 forms. The phenotypes in three sets of olfactory neurons will be assessed. And finally, single forms of Dscam will be expressed under heterologous promoters in both wild type and mutant backgrounds. Dominant phenotypes induced by these forms will be assessed. Forms that do not induce dominant phenotypes will be assessed for the ability to rescue the complete loss of function Dscam mutations. A better understanding of how neuronal connections are formed during development will provide insight into the mechanisms that may be disrupted in various neurological and psychiatric diseases.