Lift-off polymer array approach to axon guidance

在小鼠嗅觉系统中，1000万个嗅觉感觉神经元(OSNs)从上皮延伸到 嗅球。在球状体内，它们只与1800个肾小球靶点中的一个靶点和突触。这一目标 从OSNs到肾小球是一个有趣的模型系统，可以在其中研究路径发现和目标识别。 OSNs用什么分子机制来识别它们在球茎中合适的肾小球？在这 申请生物工程研究资助计划，这是一项多学科、跨领域的合作努力 神经生物学和物理学将使用新的技术来识别所使用的指导和目标线索 由OSN提供。我们假设，针对不同肾小球的OSN必须表达不同的指导线索 使他们能够区分不同的目标。为了识别这些线索，我们专注于高度开发 灵敏的分子工具。 首先，我们开发了一种能够扩增少量RNA的技术。第二，我们开发了一种 一种新颖的微阵列打印方法，大大提高了微阵列技术的灵敏度。组合在一起 一种高灵敏度的放大技术，再加上微阵列数据采集的显著改进，将 为我们提供了一个强大的屏幕来识别嗅觉系统中的指导线索。 在这项应用中，我们将进一步表征和改进这些基于聚合物的阵列的制造。我们会 将它们与传统阵列进行比较，然后使用它们来搜索由 OSN。

In the mouse olfactory system, 10 million olfactory sensory neurons (OSNs) extend from the epithelium into the olfactory bulb. Within the bulb, they will target and synapse with just one of ~1800 glomerular targets. This targeting of OSNs to glomeruli represents an intriguing model system in which to study pathfinding and target recognition. What are the molecular mechanisms used by OSNs to identify their appropriate glomerulus in the bulb? In this application to the Bioengineering Research Grant program, a multidisciplinary, collaborative effort that spans neurobiology and physics is proposed that will employ new techniques to identify guidance and targeting cues used by OSNs. We hypothesize that OSNs which target different glomeruli must express different guidance cues that enable them to distinguish among various targets. To identify these cues, we have focused on developing highly sensitive molecular tools. First, we have developed a technique capable of amplifying small amounts of RNA. Second, we have developed a novel microarray printing method that greatly improves the sensitivity of the microarray technique. The combination of a highly sensitive amplification technique coupled with a dramatic improvement in microarray data acquisition will provide us with a powerful screen to identify guidance cues in the olfactory system. In this application, we will further characterize and refine the fabrication of these polymer-based arrays. We will compare them against conventional arrays, and then use them to search for axon guidance cues expressed by OSNs.