DEVELOPMENT OF OLFACTORY CONNECTIONS

描述（由申请人提供）：该项目的主要目的是确定嗅觉系统中二尖瓣细胞树突模式的潜在机制。精确的轴突和树突模式的形成对于在大脑中建立功能性神经回路至关重要。在嗅觉系统中，嗅觉感觉神经元轴突对目标的导向机制是近年来研究的热点。然而，作为嗅神经突触后伙伴的僧帽细胞树突的分化和靶向的分子机制尚缺乏研究。我们提出了以下关于控制僧帽状/簇状细胞树突发生的假设：1）僧帽状/簇状细胞树突的形态发生需要与嗅神经的相互作用。将使用体内和体外方法测试嗅神经对二尖瓣细胞神经突延伸的影响。二尖瓣细胞树突的发育变化的特点。将使用3D胶原基质研究二尖瓣/簇状细胞树突的方向。成熟嗅觉轴突在二尖瓣细胞树突发育中的作用将使用遗传条件性消融进行检查。2)嗅神经提供可扩散的分子线索以促进树突延伸和二尖瓣/簇状神经元的分支。这一假设将直接测试使用解离嗅球培养。将使用嗅上皮条件培养基检查用于促进二尖瓣细胞树突状生长的扩散因子的存在。候选分子，脑信号蛋白，神经营养蛋白和骨形态发生蛋白将检查其树突生长促进活性在解离的嗅球神经元培养物。嗅觉上皮衍生的活性将通过凝胶过滤和使用FPLC的离子交换色谱来表征。将通过肽测序获得OE衍生活性的分子鉴别。我们研究的长期目标是了解对神经系统发育至关重要的精确细胞和分子相互作用，并深入了解发育性神经障碍和出生缺陷的原因。

DESCRIPTION (provided by applicant): The main objective of the project is to determine the underlying mechanisms that pattern the mitral cell dendrites in the olfactory system. Formation of precise axonal and dendritic pattern is crucial for establishing functional neural circuitry in the brain. In the olfactory system, the guidance mechanisms of the olfactory sensory neuron axons toward their target have been investigated recently. However, studies are still lacking for the molecular mechanisms of the differentiation and the targeting of the mitral cell dendrites, the postsynaptic partner of the olfactory nerve. We propose to investigate the following hypotheses concerning the control of mitral/tufted cell dendritogenesis: 1) Interaction with olfactory nerve is required for the morphogenesis of mitral/tufted cell dendrites. The influence of the olfactory nerve on mitral cell neurite extension will be tested using in vivo and in vitro approaches. The developmental changes of mitral cell dendrites will be characterized. The orientation of the mitral/tufted cell dendrites will be investigated using 3D collagen matrix. The role of the] mature olfactory axons in mitral cell dendritic development will be examined using genetic conditional ablation. 2) Olfactory nerve provides diffusible molecular cues to promote dendritic extension and branching of the mitral/tufted neurons. This hypothesis will be directly tested using dissociated olfactory bulb culture. The presence of the diffusible factors for promoting mitral cell dendritic growth will be examined using olfactory epithelium conditioned medium. Candidate molecules, Semaphorins, neurotrophins and BMPs will be examined for their dendrite growth promoting activity in the dissociated olfactory bulb neuron cultures. Olfactory epithelium derived activity will be characterized by gel filtration and ion exchange chromatography using FPLC. Molecular identity of the OE derived activity will be obtained by peptide sequencing. The long-term goal of our study is to understand the precise cellular and molecular interactions that are critical for the development of the nervous system and to gain insight into the causes of developmental neurological disorder and birth defects.