GENE EXPRESSION AND AUDITORY MEMORY

这个项目的目标是确定与听觉记忆有关的基因 和感知的能力。以前的工作表明， 同种形成了一个长期持久的记忆，歌曲在鸣禽的 听觉前脑NCM区与长期歌曲记忆的建立 需要mRNA和蛋白质合成在几个明确的时间 时期关于其背后复杂的分子机制知之甚少 歌曲记忆在这里，我建议使用消减基因筛选方法， 通过在NCM期间听到同种歌曲来确定NCM中调节的基因。 第一时间窗，这是歌曲呈现开始后的1-3小时。 我的初步研究已经建立了同种的歌曲刺激 这是一个可以有效诱导NCM中基因表达的范例。两个平行 将从鸟类的NCM中构建消减cDNA文库 刺激无论是同种歌曲或白色噪音，其中向上和 下调的基因通过三个循环的消减扩增来富集。 杂交方法将用对照筛选文库并诱导 probes.与两种探针差异杂交的候选基因将 通过Southern印迹分析进行鉴定、评估和测序。最后， 这些基因在NCM中的表达及其受同种鸣唱的调节 将通过定量原位杂交证实。鉴定 这些基因是重要的第一步，将导致一个新的领域， 鸣禽领域的分子研究，并最终导致 了解歌曲学习的复杂分子机制， 鸣禽的记忆和感知。

The goal of this project is to identify genes involved in auditory memory and perception in songbirds. Previous work has shown that presentation of conspecific forms a long-lasting memory of that song in a songbird's auditory forebrain area NCM and the establishment of long-term song memory requires mRNA and protein synthesis during several well defined time periods. Little is known about the complex molecular mechanisms underlying song memory. Here, I propose to use a subtractive gene screening method to identify genes regulated in NCM by hearing conspecific song during the first time window, this is 1-3 hours after the onset of song presentation. My preliminary studies have established a conspecific song stimulation paradigm that can effectively induce gene expression in NCM. Two parallel subtractive cDNA libraries will be constructed from the NCM of birds stimulated either with conspecific song or white noise, in which up and down regulated genes are enriched by three cycles of subtractive hybridization. The libraries will be screened with control and induced probes. Candidate genes differentially hybridized with the two probes will be identified, evaluated by Southern Blot analysis and sequenced. Finally, expression of these genes in NCM and their regulation by conspecific song will be confirmed by quantitative in situ hybridization. Identification of these genes is an important first step that will lead to a new area of molecular studies in songbirds field and lead to the eventual understanding of complex molecular mechanisms underlying song learning, memory and perception in songbirds.