GENE EXPRESSION AND AUDITORY MEMORY

该项目的目标是识别与听觉记忆有关的基因 以及鸣禽的感知能力。以前的工作已经表明， 在鸣鸟的歌声中形成了对那首歌的持久记忆 听觉前脑区NCM与歌曲长时记忆的建立 需要在几个明确的时间内合成mRNA和蛋白质 句号。人们对其复杂的分子机制知之甚少。 歌曲记忆。在这里，我建议使用一种消减基因筛选方法来 通过在NCM过程中听到同种歌声来识别NCM中调控的基因 第一个时间窗口，这是歌曲呈现开始后的1-3小时。 我的初步研究建立了一种同种的歌曲刺激 能有效诱导NCM基因表达的范式。两个平行的 从鸟类的NCM中构建消减cDNA文库 用同种歌曲或白噪音刺激，在这种情况下， 下调的基因通过三个周期的消减而丰富 杂交。将对文库进行对照筛选和诱导 探测器。与两个探针进行差异杂交的候选基因将 经Southern Blot分析鉴定、鉴定和测序。最后， 这些基因在NCM中的表达及同种异体SONG对它们的调节 将通过定量原位杂交进行确认。身份识别 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 了解歌曲学习背后的复杂分子机制， 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.