Concurrent Glycogen and Lactate Imaging with FTIR Spectroscopy To Spatially Localize Metabolic Parameters of the Glial Response Following Brain Ischemia
Concurrent Glycogen and Lactate Imaging with FTIR Spectroscopy To Spatially Localize Metabolic Parameters of the Glial Response Following Brain Ischemia
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DOI：
10.1021/acs.analchem.6b02588
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发表时间：
2016-11-15
影响因子：
7.4
通讯作者：
Kelly, Michael E.
中科院分区：
文献类型：
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作者：
Hackett, Mark J.;Sylvain, Nicole J.;Kelly, Michael E.
成像能量代谢产物作为脑缺血后神经胶质细胞和神经元之间能量传递的标志是一个持续的挑战。传统的显微镜结合组织化学显示脑缺血后糖原在胶质细胞内积聚，表明代谢性改变。虽然半定量组织化学糖原分析是可能的，但该方法受到组织化学常见的典型混杂因素的影响，如试剂渗透和结合的变化。此外，糖原的组织化学检测不能揭示糖原代谢命运的信息(即乳酸的产生)。因此，验证一种直接半定量方法在同一组织切片中同时成像脑糖原和乳酸将有益于这一研究领域。在这项研究中，我们首次应用傅里叶变换红外(FTIR)光谱技术在体外组织切片中同时直接成像脑糖原和乳酸。用抗胶质纤维酸性蛋白(GFAP)免疫组织化学方法对连续切片进行分析，以比较FTIR显示的糖原和乳酸分布与GFAP免疫组织化学显示的胶质分布。FTIR光谱成像揭示的糖原分布与相邻组织切片上糖原的组织化学检测作了进一步的比较。这一方法随后被应用于研究中风小鼠模型脑缺血后相对于神经胶质细胞和神经元群体的代谢时空障碍。
Imaging energy metabolites as Markers of the energy shuttle between glia and neurons following ischemia is an ongoing challenge. Traditional microscopies in combination with histochemistry reveal glycogen accumulation within glia following ischemia, indicating an altered metabolic profile. Although semiquantitative histochemical glycogen analysis is possible, the method suffers from typical confounding factors common to histochemistry, such as variation in reagent penetration and binding. In addition, histochemical detection of glycogen does not reveal information on the metabolic fate of glycogen (i.e., lactate production). Therefore, validation of a direct semiquantitative method to simultaneously image both brain glycogen and lactate in the same tissue section would benefit this research field. In this study, we demonstrate the first application of Fourier transform infrared (FTIR) spectroscopy for simultaneous direct spectroscopic imaging of brain glycogen and lactate, in situ within ex vivo tissue sections. Serial tissue sections were analyzed with anti-glial fibrillary acidic protein (GFAP) immunohistochemistry to provide a comparison between the glycogen and lactate distribution revealed by FTIR and the glial distribution revealed by GFAP immunohistochemistry. The distribution of glycogen revealed by FTIR spectroscopic imaging has been further compared with histochemical detection of glycogen on the adjacent tissue sections. This approach was then applied to study spatiotemporal disturbances in metabolism, relative to glia and neuronal populations, following cerebral ischemia in a murine model of stroke.