| Studying the Mechanisms of Developmental Vocal Learning and Adult Vocal Performance in Zebra Finches through Lentiviral Injection
|
|
Author:
Date:
2018-09-05
[Abstract] Here we provide a detailed step-by-step protocol for using lentivirus to manipulate miRNA expression in Area X of juvenile zebra finches and for analyzing the consequences on song learning and song performance. This protocol has four parts: 1) making the lentiviral construct to overexpress miRNA miR-9; 2) packaging the lentiviral vector; 3) stereotaxic injection of the lentivirus into Area X of juvenile zebra finches; 4) analysis of song learning and song performance in juvenile and adult zebra finches. These methods complement the methods employed in recent works that showed changing FoxP2 gene expression in Area X with lentivirus or adeno-associated virus leads to impairments in song behavior.
[摘要] 在这里，我们提供了一个详细的逐步协议，用于使用慢病毒操纵幼年斑胸草雀X区的miRNA表达，并分析对歌曲学习和歌曲表现的影响。 该方案有四个部分：1）使慢病毒构建体过表达miRNA miR-9; 2）包装慢病毒载体; 3）将慢病毒立体定位注入少年斑胸草雀X区; 4）青少年和成年斑马雀的歌曲学习和歌曲表演分析。 这些方法补充了近期工作中使用的方法，这些方法显示，在区域X中用慢病毒或腺相关病毒改变 FoxP2 基因表达导致歌曲行为的损害。
【背景】具有良好特征的歌曲行为和基础神经回路的斑胸草雀提供了独特的动物模型来研究声音通信和相关感觉 - 运动学习的神经机制。近年来，一些实验室开始使用病毒载体来操纵斑胸草雀脑中的基因表达并研究其功能后果。这些努力通过对 FoxP2 基因的研究得到了最好的说明，该基因编码叉头盒p2转录因子。 FoxP2蛋白控制着数百个在神经系统发育中起重要作用的下游基因的表达。人类 FoxP2 基因的突变导致言语和语言障碍（Lai et al。，2001）。在鸣禽中，斑马雀X区域的 FoxP2 基因的敲除或过表达，对于声乐学习至关重要的基底神经节核，严重损害了歌曲的行为（Haesler et al。， 2007; Murugan et al。，2013; Heston and ...
|
|
|
| 13C Tracer Studies of Metabolism in Mouse Tumor Xenografts
|
|
Author:
Date:
2015-11-20
[Abstract] Mice are widely used for human tumor xenograft studies of cancer development and drug efficacy and toxicity. Stable isotope tracing coupled with metabolomic analysis is an emerging approach for assaying metabolic network activity. In mouse models there are several routes of tracer introduction, which have particular advantages and disadvantages that depend on the model and the questions addressed. This protocol describes the bolus i.v. route via repeated tail vein injections of solutions of stable isotope enriched tracers including 13C6-glucose and 13C5,15N2-glutamine. Repeated injections give higher enrichments and over longer labeling periods than a single bolus. Multiple injections of glutamine are necessary to achieve ...
[摘要] 小鼠广泛用于癌症发展和药物功效和毒性的人肿瘤异种移植研究。 稳定同位素追踪与代谢物组学分析是一种新兴的方法来测定代谢网络活动。 在小鼠模型中，有几种示踪剂引入途径，其具有取决于模型和所解决的问题的特定优点和缺点。 该协议描述了bolus i.v. 通过重复尾静脉注射稳定的同位素富集的示踪剂的溶液的路径，所述示踪剂包括13 C 13 - 葡萄糖和13 C - 13 C - ，N 15 - ，N 15 - 谷氨酰胺。 与单次推注相比，重复注射给予更高的富集和更长的标记周期。 多次注射谷氨酰胺对于在移植肿瘤中实现充分富集是必需的。
|
|
|