The superiority and feasibility of 2,3,5-triphenyltetrazolium chloride-stained brain tissues for molecular biology experiments based on microglial properties.
The superiority and feasibility of 2,3,5-triphenyltetrazolium chloride-stained brain tissues for molecular biology experiments based on microglial properties.
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基于小胶质细胞特性的2,3,5-三苯基四唑神圣的脑组织的优势和可行性用于分子生物学实验。
DOI：
10.1002/ame2.12312
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发表时间：
2023-04
影响因子：
3.7
通讯作者：
Li, Li
中科院分区：
文献类型：
--
作者：
Wang, Yajuan;Zhang, Qingrong;Zhang, Shuchi;Qi, Jiangtao;Li, Li
TTC（氯化2，3，5-三苯基四唑）染色是在短暂性大脑中动脉闭塞模型中识别和评估脑梗死体积的最常用方法。鉴于小胶质细胞在缺血性卒中后不同区域表现出不同的形态，我们证明了基于小胶质细胞特征使用TTC染色脑组织分析不同区域各种蛋白或基因表达的优越性和必要性。我们比较了改进的TTC染色方法的脑组织（在冰上放置10分钟）与传统取样方法的半影区。我们使用真实的时间（RT）-PCR，Western blot和免疫荧光分析来确定改进染色方法的可行性和必要性。TTC染色脑组织组中无蛋白质和RNA降解。然而，TREM 2在小胶质细胞上的特异性表达在半影区显示出两组之间的显著差异。TTC染色的脑组织可用于分子生物学实验，没有任何限制。此外，TTC染色的脑组织由于其精确定位而显示出更大的优越性。TTC（2，3，5-三苯基四氮唑氯化物）染色脑组织用于基于小胶质细胞特性的分子生物学实验的优越性和可行性。
TTC (2,3,5‐triphenyltetrazolium chloride) staining is the most commonly used method in identifying and assessing cerebral infarct volumes in the transient middle cerebral artery occlusion model. Given that microglia exhibit different morphologies in different regions after ischemic stroke, we demonstrate the superiority and necessity of using TTC‐stained brain tissue to analyze the expression of various proteins or genes in different regions based on microglia character. We compared brain tissue (left for 10 min on ice) from the improved TTC staining method with penumbra from the traditional sampling method. We identified the feasibility and necessity of the improved staining method using real time (RT)‐PCR, Western blot, and immunofluorescence analysis. There was no protein and RNA degradation in the TTC‐stained brain tissue group. However, the TREM2 specifically expressed on the microglia showed a significant difference between two groups in the penumbra region. TTC‐stained brain tissue can be used for molecular biology experiments without any restrictions. In addition, TTC‐stained brain tissue shows greater superiority due to its precise positioning. The superiority and feasibility of TTC (2,3,5‐triphenyltetrazolium chloride) stained brain tissues for molecular biology experiments based on the microglial properties.