Weisz Harris A, Boone Deborah R, Sell Stacy L, Hellmich Helen L
Department of Anesthesiology, University of Texas Medical Branch.
Department of Anesthesiology, University of Texas Medical Branch;
J Vis Exp. 2017 Sep 11(127):56134. doi: 10.3791/56134.
The ability to isolate specific brain regions of interest can be impeded in tissue disassociation techniques that do not preserve their spatial distribution. Such techniques also potentially skew gene expression analysis because the process itself can alter expression patterns in individual cells. Here we describe a laser capture microdissection (LCM) method to selectively collect specific brain regions affected by traumatic brain injury (TBI) by using a modified Nissl (cresyl violet) staining protocol and the guidance of a rat brain atlas. LCM provides access to brain regions in their native positions and the ability to use anatomical landmarks for identification of each specific region. To this end, LCM has been used previously to examine brain region specific gene expression in TBI. This protocol allows examination of TBI-induced alterations in gene and microRNA expression in distinct brain areas within the same animal. The principles of this protocol can be amended and applied to a wide range of studies examining genomic expression in other disease and/or animal models.
在不保留特定脑区空间分布的组织解离技术中，分离感兴趣的特定脑区的能力可能会受到阻碍。此类技术还可能使基因表达分析产生偏差，因为该过程本身会改变单个细胞中的表达模式。在此，我们描述一种激光捕获显微切割（LCM）方法，通过使用改良的尼氏（甲酚紫）染色方案并在大鼠脑图谱的指导下，选择性收集受创伤性脑损伤（TBI）影响的特定脑区。LCM能够获取处于自然位置的脑区，并利用解剖学标志来识别每个特定区域。为此，LCM先前已被用于研究TBI中脑区特异性基因表达。本方案允许在同一动物的不同脑区中检测TBI诱导的基因和微小RNA表达变化。该方案的原理可进行修改，并应用于广泛的研究，以检测其他疾病和/或动物模型中的基因组表达。
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