Plasticity of Vocal Control

描述（由申请人提供）：本研究的长期目标是了解交流行为的神经内分泌基础。鸟类鸣叫系统是一个有价值的模型，因为鸣叫是习得的，而且它的神经回路是两性二态的，对激素敏感，在成年动物中是可塑的。鸣叫控制系统的季节性可塑性为脊椎动物大脑的可塑性提供了一个特别有趣的模型，本研究项目的一个持续目标是了解这些季节性变化的机制和功能后果。该申请的直接目标是接受分子生物学方法的培训，这将使候选人能够将他对歌曲系统可塑性的研究扩展到分子水平的分析。Brenowitz将在华盛顿大学Robert Steiner的实验室学习基因克隆、定位和靶向、单标签和双标签原位杂交、PCR和非病毒cRNA转染方法。候选人将接受西雅图Fred Hutchison癌症研究中心的Paul Neiman的培训，学习使用eDNA微阵列分析基因表达模式，以及使用生物信息学方法进行数据分析。Brenowitz将在华盛顿大学表达阵列中心学习微阵列分析课程。威斯康星大学的斯科特·爱德华兹将对布雷诺维茨进行DNA测序方法的培训。候选人将在西雅图的系统生物学研究所做“学徒”，学习蛋白质组学和生物信息学的最新方法。华盛顿大学拥有强大的研究项目，特别是在鸟鸣学、内分泌学、神经科学、分子生物学、动物行为和听力研究方面的优势。候选人在各部门的任命。心理学、动物学和布洛德特听力研究中心提供共享设施，使他的研究项目受益。本研究将探讨歌曲系统中季节性可塑性的机制和功能后果。目标1和目标2将测试季节性生长的社会增强是由听觉线索介导的，并涉及增加的神经元招募。目的3将测试传入神经支配是维持季节性生长的歌核所必需的假设。目的4将验证歌曲回路的季节性生长是由睾酮的雌激素代谢物介导的假设。目的5将使用操作性条件反射来检验假设，即歌曲核的季节性可塑性导致歌曲感知的季节性变化。Aim 6将使用原位杂交技术来验证脑源性神经营养因子基因表达在歌唱回路季节性生长时上调的假设。Aim 7将使用减法抑制杂交来识别在季节性生长的细胞核中富集的基因，与不生长的细胞核相比。Aim 8将使用定制的eDNA微阵列来分析与端脑歌唱控制回路季节性生长相关的基因表达的全局模式。这些研究结果将增加我们对类固醇激素和社会刺激对神经系统的影响的理解，以及成人大脑可塑性与学习之间的关系。

DESCRIPTION (provided by applicant): The long-term goal of this research is to understand the neuroendocrine basis of behavior used in communication. The birdsong system is a valuable model because song is learned, and its neural circuits are sexually dimorphic, hormone sensitive, and plastic in adults. Seasonal plasticity of the song control system provides an especially interesting model of plasticity in vertebrate brains, and a continuing goal of this research program is to understand the mechanisms and functional consequences of these seasonal changes. The immediate goal of this application is to receive training in methods of molecular biology that will enable the candidate to extend his studies of plasticity of the song system to a molecular level of analysis. Brenowitz will learn gene cloning, mapping and targeting, single and double-label in situ hybridization, PCR, and nonviral cRNA transfection methods in the laboratory of Robert Steiner at the University of Washington (UW). The candidate will be trained by Paul Neiman of the Fred Hutchison Cancer Research Center in Seattle in the use of eDNA microarrays to analyze patterns of gene expression, and in bioinformatic methods for data analysis. Brenowitz will take a course on microarray analysis at the UW Center for Expression Arrays. Scott Edwards of UW will train Brenowitz in DNA sequencing methods. The candidate will "apprentice" at the Institute for Systems Biology in Seattle to learn current methods of proteomics and bioinformatics. UW has a strong research program, with particular strengths in birdsong, endocrinology, neuroscience, molecular biology, animal behavior, and hearing research. The candidate's appointments in the Depts. of Psychology, Zoology, and the Bloedet Hearing Research Center provide access to shared facilities that benefit his research program. The proposed research will address mechanisms and functional consequences of seasonal plasticity in the song system. Aims 1 & 2, will test the hypotheses that social enhancement of seasonal growth is mediated by auditory cues and involves increased neuronal recruitment. Aim 3 will test the hypothesis that afferent innervation is necessary to maintain seasonally grown song nuclei. Aim 4 will test the hypothesis that seasonal growth of song circuits is mediated by estrogenic metabolites of testosterone. Aim 5 will use operant conditioning to test the hypothesis that seasonal plasticity of the song nuclei causes seasonal changes in song perception. Aim 6 will use in situ hybridization to test the hypothesis that expression of the gene for brain derived neurotrophic factor is upregulated when the song circuits are seasonally growing. Aim 7 will use subtractive suppressive hybridization to identify genes that are enriched in a song nucleus that grows seasonally compared with a nucleus that does not grow. Aim 8 will use a customized eDNA microarray to analyze global patterns of gene expression that are associated with seasonal growth of the telencephalic song control circuits. The results of the proposed studies will increase our understanding of the influences of steroid hormones and social stimuli on the nervous system, and the relationship between plasticity in the adult brain and learning.