Development of the calyx of Held

描述（由申请人提供）：参与突触后靶点输入之间竞争的因素，获胜输入的生长和稳定，以及神经支配在突触伴侣协调成熟中的作用尚未得到很好的描述。在某种程度上，这种情况源于大多数神经系统中这些事件的长期性质。我们已经发现，神经支配的细胞在听觉脑干的大神经末梢称为杯的赫尔德是一个很好的模型系统来研究这些事件。小的终端竞争，然后一个单一的获胜输入增长到48小时内，包裹其目标的细胞体，出生后（P）天之间2-4。我们将利用这个有限的时间框架，研究使用微阵列技术的基因表达的起始和偏移。我们建议在这里分析数据，从密集的时间采样的基因表达周围P2-4在每天的时间间隔使用外显子微阵列。从基因的个体表达谱在时间上与P2-4的突触发生事件相关，我们将选择25个候选者，通过使用定量实时PCR，免疫组织化学和原位杂交进行验证。更复杂的分析，如基因簇或基于剪接的替代方法，将产生一个额外的基因列表，从中选择另外25个候选基因进行验证。这些活动将需要征聘人员填补两个新职位。通过与约翰霍普金斯大学的蒋谦博士的新合作，将聘请一名博士后研究员协助钱博士进行微阵列分析。在西弗吉尼亚大学，将聘请一名研究技术人员，以促进选定的候选基因的高通量验证。这些新创造的工作岗位，沿着更有效地进行qPCR实验所需的设备，将加快科学研究的克里思。本申请是根据拨款通知编号：NOT-09-058和通知标题：NIH宣布恢复法基金的竞争力修订申请的可用性提交的。 公共卫生相关性：大脑中的信息处理障碍表征了一系列神经病理学，这些神经病理学可以追溯到早期发育期间神经回路的不正确形成。拟议的研究旨在发现神经回路形成的一般原则。因此，这项工作的见解可能会支持听力缺陷的治疗策略，如听觉处理障碍和阅读障碍。

DESCRIPTION (provided by applicant): The factors involved in competition among inputs for a postsynaptic target, growth and stabilization of winning inputs, and the role of innervation in coordinated maturation of synaptic partners are not well delineated. In part this situation stems from the protracted nature of these events in most neural systems. We have revealed that innervation of cells in the auditory brainstem by large nerve terminals called calyces of Held is a good model system to study these events. Small terminals compete, then a single winning input grows to envelop the cell body of its target within 48 hours, between postnatal (P) days 2-4. We will take advantage of this constrained time frame to study onset and offset of gene expression using microarray technology. We propose here to analyze data from dense temporal sampling of gene expression surrounding P2-4 in daily intervals using exon microarrays. From genes whose individual expression profile is temporally linked to synaptogenesis events at P2-4, we will select 25 candidates for validation by using quantitative real-time PCR, immunohistochemistry and in situ hybridization. More complex analyses, such as gene cluster or alternative splicing-based methods, will yield an additional list of genes from which another 25 candidates will be selected for validation. These activities will require recruitment to fill two new positions. Through a new collaboration with Jiang Qian, PhD, at Johns Hopkins University, a postdoctoral fellow will be hired to assist Dr. Qian with microarray analyses. At West Virginia University, a research technician will be hired to facilitate high throughput validation of selected candidate genes. These newly created jobs, along with requested equipment to more efficiently perform qPCR experiments, will accelerate the tempo of scientific research. This application is submitted in accordance with grant Notice Number: NOT-09-058 and Notice Title: NIH Announces the Availability of Recovery Act Funds for Competittivce Revision Applications. PUBLIC HEALTH RELEVANCE: Disorders of information processing in the brain characterize a range of neurologic pathologies that can be traced to improper formation of neural circuits during early development. The proposed studies are designed to discover general principles that underlie neural circuit formation. Therefore, insights from this work may underpin therapeutic strategies for deficits in hearing, such as auditory processing disorder and dyslexia.