Platt Virginia, Lee Do Yup, Canaria Christie A, Frankel Ken, Bernstein Susan, McMurray Cynthia T
Lawrence Berkeley National laboratory, Berkeley, CA, USA.
Methods Mol Biol. 2013;1010:213-30. doi: 10.1007/978-1-62703-411-1_14.
Many trinucleotide repeat disorders exhibit region-specific toxicity within tissues, the basis of which cannot be explained by traditional methods. For example, in Huntington's Disease (HD), the toxic disease-causing protein is ubiquitously expressed. However, only the medium spiny neurons in the striatum are initially targeted for death. Many changes are likely to initiate in these cells at an intracellular and microstructural level long before there is a measureable phenotype, but why some regions of the brain are more susceptible to death is unknown. This chapter describes a method to detect functional changes among brain regions and cell types, and link them directly with region-specific physiology. Due to the neurodegeneration that accompanies many triplet repeat disorders, we focus on the brain, although the methods described in this chapter can be translated to other tissue types. We integrate immunohistology and traditional mass spectrometry with a novel mass spectrometry imaging technique, called nanostructure initiated mass spectrometry (NIMS). When used together, these tools offer unique insights into region-specific physiology of the brain, and a basis for understanding the region-specific toxicity associated with triplet repeat disorders.
许多三核苷酸重复序列疾病在组织内表现出区域特异性毒性，其原因无法用传统方法解释。例如，在亨廷顿舞蹈症（HD）中，有毒的致病蛋白在全身各处表达。然而，最初只有纹状体中的中型多棘神经元会成为死亡目标。在出现可测量的表型之前很久，这些细胞在细胞内和微观结构水平上可能就已经发生了许多变化，但大脑的某些区域为何更容易死亡尚不清楚。本章介绍了一种检测脑区和细胞类型之间功能变化，并将它们直接与区域特异性生理学联系起来的方法。由于许多三联体重复序列疾病都伴有神经退行性变，我们将重点放在大脑上，不过本章所述方法也可应用于其他组织类型。我们将免疫组织学和传统质谱技术与一种名为纳米结构引发质谱（NIMS）的新型质谱成像技术相结合。这些工具一起使用时，能为大脑的区域特异性生理学提供独特见解，并为理解与三联体重复序列疾病相关的区域特异性毒性奠定基础。
