TREX1 is expressed by microglia in normal human brain and increases in regions affected by ischemia.
TREX1 is expressed by microglia in normal human brain and increases in regions affected by ischemia.
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DOI：
10.1111/bpa.12626
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发表时间：
2018-11
期刊：
影响因子：
--
通讯作者：
Atkinson JP
中科院分区：
文献类型：
--
作者：
Kothari PH;Kolar GR;Jen JC;Hajj-Ali R;Bertram P;Schmidt RE;Atkinson JP
三素修复核酸外切酶1（TREX1）基因的突变与神经系统疾病相关，包括视网膜血管病变伴脑白质脑病（RVCL）。然而，TREX1在人脑中的内源性表达尚未研究。我们制备了一种针对TREX1的兔多克隆抗体（pAb），以通过来自正常对照和携带RVCL移码突变的受试者的细胞裂解物的蛋白质印迹（WB）来表征TREX1。进行双重染色以确定人脑组织中表达TREX1的细胞类型。通过对来自正常对照和RVCL和缺血性卒中患者的福尔马林固定、石蜡包埋样本进行免疫组织化学分析，进一步评估人脑中的TREX1分布。在验证我们的抗TREX1兔pAb的特异性后，使用WB分析来检测细胞裂解物中TREX1的内源性野生型和移码突变体。在来自RVCL患者和正常对照的人脑组织中的双重染色将TREX 1定位于小胶质细胞和巨噬细胞的子集。大脑皮质免疫组织化学染色的定量显示，TREX 1+小胶质细胞主要在正常对照的灰质中（在灰质和白色物质中分别有22.7 ± 5.1%和5.5 ± 1.9%的Iba 1+小胶质细胞），并且通常与微血管相关。相比之下，在RVCL受试者中，TREX 1+小胶质细胞主要位于正常外观的大脑皮质的白色物质中（Iba 1+小胶质细胞的11.8 ± 3.1%和38.9 ± 5.8%分别位于灰色和白色物质中）。RVCL和脑卒中患者中枢神经系统缺血性病变中TREX1+小胶质细胞数量增加。 TREX1由正常人脑中的小胶质细胞亚群表达，通常与微血管非常接近，并且在缺血性病变的情况下增加。这些发现表明TREX1+小胶质细胞在血管稳态和对缺血性损伤的反应中的作用。
Mutations in the three‐prime repair exonuclease 1 (TREX1) gene have been associated with neurological diseases, including Retinal Vasculopathy with Cerebral Leukoencephalopathy (RVCL). However, the endogenous expression of TREX1 in human brain has not been studied. We produced a rabbit polyclonal antibody (pAb) to TREX1 to characterize TREX1 by Western blotting (WB) of cell lysates from normal controls and subjects carrying an RVCL frame‐shift mutation. Dual staining was performed to determine cell types expressing TREX1 in human brain tissue. TREX1 distribution in human brain was further evaluated by immunohistochemical analyses of formalin‐fixed, paraffin‐embedded samples from normal controls and patients with RVCL and ischemic stroke. After validating the specificity of our anti‐TREX1 rabbit pAb, WB analysis was utilized to detect the endogenous wild‐type and frame‐shift mutant of TREX1 in cell lysates. Dual staining in human brain tissues from patients with RVCL and normal controls localized TREX1 to a subset of microglia and macrophages. Quantification of immunohistochemical staining of the cerebral cortex revealed that TREX1+ microglia were primarily in the gray matter of normal controls (22.7 ± 5.1% and 5.5 ± 1.9% of Iba1+ microglia in gray and white matter, respectively) and commonly in association with the microvasculature. In contrast, in subjects with RVCL, the TREX1+ microglia were predominantly located in the white matter of normal appearing cerebral cortex (11.8 ± 3.1% and 38.9 ± 5.8% of Iba1+ microglia in gray and white matter, respectively). The number of TREX1+ microglia was increased in ischemic brain lesions in central nervous system of RVCL and stroke patients. TREX1 is expressed by a subset of microglia in normal human brain, often in close proximity to the microvasculature, and increases in the setting of ischemic lesions. These findings suggest a role for TREX1+ microglia in vessel homeostasis and response to ischemic injury.
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