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（OSN）从上皮细胞延伸到嗅觉神经元。 嗅球在球内，它们将靶向约1800个肾小球靶点中的一个并与之突触。该靶向 OSN的肾小球代表了一个有趣的模型系统，在其中研究寻路和目标识别。 OSN使用什么样的分子机制来识别球中合适的肾小球？在这 申请生物工程研究资助计划，这是一个跨学科的合作项目， 神经生物学和物理学的建议，将采用新的技术，以确定指导和目标线索使用 的OSN。我们假设靶向不同肾小球的OSN必须表达不同的引导信号， 使他们能够区分不同的目标。为了识别这些线索，我们专注于开发高度 敏感的分子工具 首先，我们开发了一种能够扩增少量RNA的技术。第二，我们制定了 一种新的微阵列打印方法，大大提高了微阵列技术的灵敏度。相结合 高灵敏度的扩增技术与微阵列数据采集的显著改进相结合， 为我们提供了一个强大的屏幕来识别嗅觉系统中的引导线索。 在本申请中，我们将进一步表征和完善这些聚合物基阵列的制造。我们将 将它们与传统阵列进行比较，然后用它们来搜索轴突引导线索， 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.