HMGB1 Translocation in Neurons after Ischemic Insult: Subcellular Localization in Mitochondria and Peroxisomes
HMGB1 Translocation in Neurons after Ischemic Insult: Subcellular Localization in Mitochondria and Peroxisomes
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DOI：
10.3390/cells9030643
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发表时间：
2020-03
期刊：
影响因子：
6
通讯作者：
Dengli Wang;Keyue Liu;Yusuke Fukuyasu;K. Teshigawara;Li Fu;H. Wake;A. Ohtsuka;M. Nishibori
中科院分区：
文献类型：
--
作者：
Dengli Wang;Keyue Liu;Yusuke Fukuyasu;K. Teshigawara;Li Fu;H. Wake;A. Ohtsuka;M. Nishibori
高迁移率族蛋白-1（HMGB 1）是一种非组蛋白染色质DNA结合蛋白，在缺血、出血和创伤损伤时从神经元释放到细胞外间隙。然而，HMGB 1的时间依赖性易位和HMGB 1通过神经元释放过程的亚细胞定位的细节仍不清楚。在本研究中，我们研究了HMGB 1的亚细胞定位过程中易位的HMGB 1在胞质隔室使用大脑中动脉闭塞和再灌注模型大鼠。免疫荧光双标显微镜显示HMGB 1与MTCO 1共定位（线粒体编码的细胞色素c氧化酶I），线粒体的标志物，和过氧化氢酶，过氧化物酶体的标志物，但不与Rab 5/Rab 7（RAS相关GTP结合蛋白），LC 3 A/B（微管相关蛋白1轻链3）（KDEL氨基酸序列）和LAMP 1（溶酶体相关膜蛋白1），它们分别是核内体、吞噬体、内质网和溶酶体标志物。免疫电子显微镜证实，免疫金颗粒HMGB 1的线粒体和过氧化物酶体内。此外，发现HMGB 1与参与线粒体分裂的Drp 1（动力蛋白相关蛋白1）共定位。这些结果揭示了HMGB 1在缺血条件下释放过程中的特异性亚细胞定位。
High mobility group box-1 (HMGB1), a nonhistone chromatin DNA-binding protein, is released from neurons into the extracellular space under ischemic, hemorrhagic, and traumatic insults. However, the details of the time-dependent translocation of HMGB1 and the subcellular localization of HMGB1 through the release process in neurons remain unclear. In the present study, we examined the subcellular localization of HMGB1 during translocation of HMGB1 in the cytosolic compartment using a middle cerebral artery occlusion and reperfusion model in rats. Double immunofluorescence microscopy revealed that HMGB1 immunoreactivities were colocalized with MTCO1(mitochondrially encoded cytochrome c oxidase I), a marker of mitochondria, and catalase, a marker of peroxisomes, but not with Rab5/Rab7 (RAS-related GTP-binding protein), LC3A/B (microtubule-associated protein 1 light chain 3), KDEL (KDEL amino acid sequence), and LAMP1 (Lysosomal Associated Membrane Protein 1), which are endosome, phagosome, endoplasmic reticulum, and lysosome markers, respectively. Immunoelectron microscopy confirmed that immune-gold particles for HMGB1 were present inside the mitochondria and peroxisomes. Moreover, HMGB1 was found to be colocalized with Drp1 (Dynamin-related protein 1), which is involved in mitochondrial fission. These results revealed the specific subcellular localization of HMGB1 during its release process under ischemic conditions.