IMAGING AXON GROWTH CONE/GLIA INTERACTIONS IN THE DEVELOPING OLFACTORY SYSTEM

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 嗅觉系统轴突生长锥/胶质细胞相互作用的成像研究 我们专注于在我们的实验室发展轴突和神经胶质细胞之间的相互作用，了解神经胶质细胞在轴突导航到他们的目标在中枢神经系统的作用。 我们想表征轴突生长锥的行为过程中的轴突分类在一个区域的嗅觉通路，是由神经胶质细胞和嗅觉受体轴突排序从其他雅阁与他们的目标区域在嗅叶。我们开发了一种器官型制剂，包括蛾天蛾的整个主要嗅觉通路，并保持3D环境，通过该环境，受体轴突通常延伸，使我们能够分析轴突生长锥的行为，因为它们在神经胶质细胞附近延伸或接触。 这些数据作为实验的基础，在这些实验中，当胶质细胞数量减少或分子环境改变时，我们可以跟踪生长锥的行为。 由于我们目前使用标准共聚焦显微镜对生长中的轴突进行延时成像的特点是光漂白，光毒性和有限的深度分辨率的挑战，我们需要专家帮助用多光子系统对我们的制备进行成像，以获取初步数据并优化我们使用这种系统的协议。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. IMAGING AXON GROWTH CONE/GLIA INTERACTIONS IN THE DEVELOPING OLFACTORY SYSTEM We focus in our laboratory on developmental interactions between growing axons and glial cells with the goal of understanding the role of glia in axon navigation toward their targets in the CNS. We wanted to characterize axonal growth cone behavior during the process of axon sorting in a region of the olfactory pathway that is populated by glial cells and in which olfactory receptor axons sort from one other in accord with their target region in the olfactory lobe. We developed an organotypic preparation that comprises the entire primary olfactory pathway of the moth Manduca sexta and maintains the 3D environment through which the receptor axons normally extend to allow us to analyze the behavior of axonal growth cones as they extend near or in contact with the glia. These data serve as the foundation for experiments in which we can follow growth cone behavior when glial number is reduced or when the molecular environment in altered. Because our current time-lapse imaging of growing axons using standard confocal microscopy is marked by challenges of photo-bleaching, photo-toxicity and limited depth resolution, we needed expert assistance in imaging our preparation with a multi-photon system, both to acquire preliminary data and to optimize our protocols for working with such a system.