Winrow Christopher J, Tanis Keith Q, Rigby Alison M, Taylor Rhonda R, Serikawa Kyle, McWhorter Mollie, Tokiwa George Y, Marton Matthew J, Stone David J, Koblan Kenneth S, Renger John J
Depression and Circadian Disorders Department, Merck Research Laboratories, West Point, PA 19486, USA.
Brain Res. 2009 May 19;1271:1-17. doi: 10.1016/j.brainres.2009.02.083. Epub 2009 Mar 17.
Powerful new approaches to study molecular variation in distinct neuronal populations have recently been developed enabling a more precise investigation of the control of neural circuits involved in complex behaviors such as wake and sleep. We applied laser capture microdissection (LCM) to isolate precise brain nuclei from rat CNS at opposing circadian time points associated with wake and sleep. Discrete anatomical and temporal analysis was performed to examine the extent of variation in the transcriptional control associated with both identifiable anatomical nuclei and with light/dark cycle. Precise isolation of specific brain nuclei regulating sleep and arousal, including the LC, SCN, TMN, VTA, and VLPO, demonstrated robust changes in gene expression. Many of these differences were not observed in previous studies where whole brain lysates or gross dissections were used to probe for changes in gene expression. The robust and differential profiles of genomic data obtained from the approaches used herein underscore the requirement for careful anatomical refinement in CNS gene expression studies designed to understand genomic control within behaviorally-linked, but functionally isolated brain nuclei.
最近开发了强大的新方法来研究不同神经元群体中的分子变异，从而能够更精确地研究参与诸如清醒和睡眠等复杂行为的神经回路的控制。我们应用激光捕获显微切割技术（LCM），在与清醒和睡眠相关的相反昼夜节律时间点，从大鼠中枢神经系统中分离出精确的脑核。进行了离散的解剖学和时间分析，以检查与可识别的解剖学核以及光/暗周期相关的转录控制的变化程度。对调节睡眠和觉醒的特定脑核（包括蓝斑、视交叉上核、结节乳头体核、腹侧被盖区和腹外侧视前区）的精确分离，显示出基因表达的强烈变化。在以前使用全脑裂解物或大体解剖来探测基因表达变化的研究中，并未观察到许多这些差异。本文所用方法获得的基因组数据的强大且差异显著的图谱，强调了在旨在了解行为相关但功能孤立的脑核内基因组控制的中枢神经系统基因表达研究中，进行仔细解剖细化的必要性。
