The effects of postmortem delay on mouse and human microglia gene expression.
The effects of postmortem delay on mouse and human microglia gene expression.
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作者：
Heng Y;Dubbelaar ML;Marie SKN;Boddeke EWGM;Eggen BJL
小胶质细胞是中枢神经系统（CNS）的特化巨噬细胞，并且首先对病原体或损伤作出反应。在过去的十年中，小胶质细胞的转录谱显着有助于我们了解他们的功能。在人CNS样本的情况下，手术样本情况下的潜在CNS病理学或由于组织获取和收集所需的时间导致的死后延迟（PMD）是影响基因表达谱的潜在因素。为了确定PMD对小胶质细胞转录组的影响，我们首先分析了小鼠小胶质细胞，其中基因型、死前条件和PMD可以控制。使用荧光激活细胞分选（FACS）从不同PMD（0、4、6、12和24小时）后的小鼠中分离小胶质细胞。存活的小胶质细胞的数量随着PMD的增加而显著减少，但即使在12小时PMD后，也可以获得高质量的RNA。PMD对小鼠小胶质细胞基因表达的影响非常有限，只有50个基因在不同PMD之间差异表达。这些基因与线粒体、核糖体和蛋白质结合功能有关。在我们之前生成的人类小胶质细胞转录组中，50个PMD相关小鼠基因中有31个具有人类同源物，并且它们的相对表达也受到PMD的影响。这项研究提供了一组基因，显示出与PMD相关的相对表达变化，在小鼠和人类小胶质细胞中。虽然检测到的基因表达变化是微妙的，这些基因需要考虑时，分析小胶质细胞转录组产生的样本与变量PMD。死后延迟在有限程度上影响小胶质细胞基因表达。死后延迟对小鼠和人类小胶质细胞转录组的影响是相当的。受影响的基因的表达随着死后延迟的增加而减少。
Microglia are specialized macrophages of the central nervous system (CNS) and first to react to pathogens or injury. Over the last decade, transcriptional profiling of microglia significantly contributed to our understanding of their functions. In the case of human CNS samples, either potential CNS pathology in the case of surgery samples, or a postmortem delay (PMD) due to the time needed for tissue access and collection, are potential factors that affect gene expression profiles. To determine the effect of PMD on the microglia transcriptome, we first analyzed mouse microglia, where genotype, antemortem conditions and PMD can be controlled. Microglia were isolated from mice after different PMDs (0, 4, 6, 12, and 24 hr) using fluorescence‐activated cell sorting (FACS). The number of viable microglia significantly decreased with increasing PMD, but even after a 12 hr PMD, high‐quality RNA could be obtained. PMD had very limited effect on mouse microglia gene expression, only 50 genes were differentially expressed between different PMDs. These genes were related to mitochondrial, ribosomal, and protein binding functions. In human microglia transcriptomes we previously generated, 31 of the 50 PMD‐associated mouse genes had human homologs, and their relative expression was also affected by PMD. This study provides a set of genes that shows relative expression changes in relation to PMD, both in mouse and human microglia. Although the gene expression changes detected are subtle, these genes need to be accounted for when analyzing microglia transcriptomes generated from samples with variable PMDs. Postmortem delay affects microglia gene expression to a limited extent. The effect of postmortem delay on the mouse and human microglia transcriptome is comparable. Affected genes decrease in expression with increasing postmortem delay.
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影响因子：
25
作者：
Galatro, Thais F.;Holtman, Inge R.;Eggen, Bart J. L.
通讯作者：
Eggen, Bart J. L.
影响因子：
2.1
作者：
Heinrich, Marielle;Matt, Katja;Schmidt, Ulrike
通讯作者：
Schmidt, Ulrike
DOI：
10.1007/978-1-4939-6786-5_23
发表时间：
2017-01-01
期刊：
INFLAMMATION: METHODS AND PROTOCOLS
影响因子：
--
作者：
Galatro, Thais F.;Vainchtein, Ilia D.;Eggen, Bart J. L.
通讯作者：
Eggen, Bart J. L.
DOI：
10.1097/nen.0b013e31815c196a
发表时间：
2007-12-01
影响因子：
3.2
作者：
Ervin, John F.;Heinzen, Erin L.;Hulette, Christine M.
通讯作者：
Hulette, Christine M.
影响因子：
2.1
作者：
Koppelkamm, Antje;Vennemann, Benedikt;Vennemann, Marielle
通讯作者：
Vennemann, Marielle