Receptor activator of nuclear factor-kappa B is enriched in CD9-positive extracellular vesicles released by osteoclasts.
Receptor activator of nuclear factor-kappa B is enriched in CD9-positive extracellular vesicles released by osteoclasts.
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DOI：
10.20517/evcna.2023.38
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Holliday, Lexie Shannon
中科院分区：
文献类型：
--
作者：
Ruan, Shaobo;Rody Jr, Wellington J.;Patel, Shivani S.;Hammadi, Lina I.;Martin, Macey L.;de Faria, Lorraine P.;Daaboul, George;Anderson, Leif S.;He, Mei;Holliday, Lexie Shannon
关键词：
核因子κ B受体激活剂（Receptor activator of nuclear factor-kappa B， RANK）-containing extracellular vesicles， EVs与成骨细胞上的RANK-配体（RANK - ligand， RANKL）结合，从而同时抑制骨吸收和促进骨形成。正因为如此，它们是有吸引力的候选治疗骨合成代谢剂。以前，通过免疫金电镜，每36个破骨细胞中就有1个检测到RANK。在这里，我们试图更详细地描述含有RANK的破骨细胞的ev亚群。免疫荧光法检测破骨细胞中四跨蛋白CD9和CD81的定位。透射电镜观察ev。单粒子干涉反射成像传感器（SP-IRIS）和免疫亲和分离检测RANK是否在特定类型的ev中富集。免疫荧光显示CD9主要位于破骨细胞的质膜上或附近。相比之下，CD81在破骨细胞的胞浆泡网络中定位较深。通过干涉测量，来自破骨细胞的CD9和CD81阳性ev都很小（直径56-83 nm），与电子显微镜一致。CD9和CD81 EV群体大多是不同的，只有22%的EV包含这两种标记。SP-IRIS在2%-4%的含cd9的成熟破骨细胞中检测到RANK，但在cd81阳性的成熟破骨细胞中未检测到RANK。从破骨细胞条件培养基中免疫磁分离含cd9的ev可去除大部分RANK。用CD81分离出微量RANK。在cd9阳性ev的一个子集中，RANK富集。目前的研究首次报道了电动汽车亚群中RANK的选择性定位。
Receptor activator of nuclear factor-kappa B (RANK)-containing extracellular vesicles (EVs) bind RANK-Ligand (RANKL) on osteoblasts, and thereby simultaneously inhibit bone resorption and promote bone formation. Because of this, they are attractive candidates for therapeutic bone anabolic agents. Previously, RANK was detected in 1 in every 36 EVs from osteoclasts by immunogold electron microscopy. Here, we have sought to characterize the subpopulation of EVs from osteoclasts that contains RANK in more detail. The tetraspanins CD9 and CD81 were localized in osteoclasts by immunofluorescence. EVs were visualized by transmission electron microscopy. A Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS) and immunoaffinity isolations examined whether RANK is enriched in specific types of EVs. Immunofluorescence showed CD9 was mostly on or near the plasma membrane of osteoclasts. In contrast, CD81 was localized deeper in the osteoclast’s cytosolic vesicular network. By interferometry, both CD9 and CD81 positive EVs from osteoclasts were small (56-83 nm in diameter), consistent with electron microscopy. The CD9 and CD81 EV populations were mostly distinct, and only 22% of the EVs contained both markers. RANK was detected by SP-IRIS in 2%-4% of the CD9-containing EVs, but not in CD81-positive EVs, from mature osteoclasts. Immunomagnetic isolation of CD9-containing EVs from conditioned media of osteoclasts removed most of the RANK. A trace amount of RANK was isolated with CD81. RANK was enriched in a subset of the CD9-positive EVs. The current study provides the first report of selective localization of RANK in subsets of EVs.
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DOI：
10.20517/evcna.2020.02
发表时间：
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期刊：
Extracellular vesicles and circulating nucleic acids
影响因子：
--
作者：
Holliday LS;Patel SS;Rody WJ Jr
通讯作者：
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影响因子：
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DOI：
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发表时间：
2007-12-01
期刊：
Allergology international : official journal of the Japanese Society of Allergology
影响因子：
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作者：
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