SPECIFIC BINDING OF A MUTATED FRAGMENT OF CLOSTRIDIUM PERFRINGENS ENTEROTOXIN TO ENDOTHELIAL CLAUDIN-5 AND ITS MODULATION OF CEREBRAL VASCULAR PERMEABILITY
SPECIFIC BINDING OF A MUTATED FRAGMENT OF CLOSTRIDIUM PERFRINGENS ENTEROTOXIN TO ENDOTHELIAL CLAUDIN-5 AND ITS MODULATION OF CEREBRAL VASCULAR PERMEABILITY
复制标题
产气荚膜梭菌肠毒素突变片段与内皮claudin-5的特异性结合及其对脑血管通透性的调节
DOI：
10.1016/j.neuroscience.2016.04.013
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发表时间：
2016-07-07
期刊：
影响因子：
3.3
通讯作者：
Zhang, Jingjing
中科院分区：
文献类型：
--
作者：
Liao, Zhuangbin;Yang, Zhenguo;Zhang, Jingjing
脊椎动物血脑屏障（BBB）对中枢神经系统相关的药物递送造成障碍。Claudin-5（Cldn 5）在血脑屏障中大量表达，在限制血脑屏障通透性方面起重要作用。产气荚膜梭菌肠毒素（cCPE）的C-末端结构域已被验证为与封闭蛋白（Cldns）的子集结合。在本研究中应用Cldn 5结合cCPE（194-319）变体cCPE（Y306 W/S313 H）来研究其调节斑马鱼幼虫血脑屏障通透性的能力。体外实验结果表明，cCPE（Y306 W/S313 H）能够特异性结合小鼠脑血管内皮细胞（弯曲. 3)细胞，并与Cldn 5一起沿着从细胞膜转运到细胞质，这又导致跨内皮电阻（TEER）的降低。相反，这种效果可以通过去除cCPE（Y306 W/S313 H）来逆转。在体内实验中，本研究使用罗丹明B-葡聚糖染料扩散测定法在斑马鱼幼虫BBB中估计cCPE（Y306 W/S313 H）调节Cldn 5的能力。结果表明，cCPE（Y306 W/S313 H）与Cldn 5在斑马鱼脑血管细胞中共定位，并调节BBB通透性，导致染料渗漏。总之，该研究表明cCPE（Y306 W/S313 H）具有通过特异性结合Cldn 5暂时调节BBB通透性的能力-在体外和体内。(C)2016年IBRO。由爱思唯尔有限公司出版。保留所有权利。
The vertebrate blood-brain barrier (BBB) creates an obstacle for central nervous system-related drug delivery. Claudin-5 (Cldn5), expressed in large quantities in BBB, plays a vital role in restricting BBB permeability. The C-terminal domain of Clostridium perfringens enterotoxin (cCPE) has been verified as binding to a subset of claudins (Cldns). The Cldn5-binding cCPE(194-319) variant cCPE(Y306W/S313H) was applied in this study to investigate its ability to modulate the permeability of zebrafish larval BBB. In vitro results showed that cCPE(Y306W/S313H) is able to bind specifically to Cldn5 in murine brain vascular endothelial (bEnd. 3) cells, and is transported along with Cldn5 from the cell membrane to the cytoplasm, which in turn results in a reduction in transendothelial electrical resistance (TEER). Conversely, this effect can be reversed by removal of cCPE(Y306W/S313H). In an in vivo experiment, this study estimates the capability of cCPE(Y306W/S313H) to modulate Cldn5 using a rhodamine B-Dextran dye diffusion assay in zebrafish larval BBB. The results show that cCPE(Y306W/S313H) co-localized with Cldn5 in zebrafish cerebral vascular cells and modulated BBB permeability, resulting in dye leakage. Taken together, this study suggests that cCPE(Y306W/S313H) has the capability - both in vitro and in vivo - to modulate BBB permeability temporarily by specific binding to Cldn5. (C) 2016 IBRO. Published by Elsevier Ltd. All rights reserved.