Brunner Nora, Stein Laura, Cornelius Valeria, Knittel Ria, Fallier-Becker Petra, Amasheh Salah
Department of Veterinary Medicine, Institute of Veterinary Physiology, Freie Universität Berlin, Berlin, Germany.
Institute of Pathology and Neuropathology, University Hospital of Tuebingen, Eberhard Karls University of Tuebingen, Tuebingen, Germany.
Front Physiol. 2020 Jul 21;11:857. doi: 10.3389/fphys.2020.00857. eCollection 2020.
Claudin-5 determines the sealing properties of blood-brain barrier tight junctions and its function is impaired in neurodegenerative and neuroinflammatory disorders. Focusing on the contribution of claudin-5 to the -interaction within the tight junction seal, we used oocytes as an expression system. Cells were clustered and challenged in a novel approach for the analysis of claudin interaction. We evaluated the strengthening effect of claudin-5 to cell-cell-connection in comparison to claudin-3. Application of a hydrostatic pressure impulse on clustered control oocyte pairs revealed a reduction of contact areas. In contrast, combinations with both oocytes expressing claudins maintained an enhanced connection between the cells (cldn5-cldn5, cldn3-cldn3). Strength of interaction was increased by both claudin-3 and claudin-5. This novel approach allowed an analysis of single claudins contributing to tight junction integrity, characterizing homophilic and hetrophilic -interaction of claudins. To test a new screening approach for barrier effectors, exemplarily, this 2-cell model of oocytes was used to analyze the effect of the absorption enhancer sodium caprate on the oocyte pairs.
Claudin-5决定血脑屏障紧密连接的封闭特性，其功能在神经退行性和神经炎症性疾病中受损。聚焦于Claudin-5对紧密连接密封内相互作用的贡献，我们使用卵母细胞作为表达系统。细胞被聚集并采用一种新方法进行挑战，以分析Claudin的相互作用。与Claudin-3相比，我们评估了Claudin-5对细胞间连接的增强作用。对聚集的对照卵母细胞对施加静水压力脉冲显示接触面积减小。相反，两个表达Claudin的卵母细胞组合可维持细胞间增强的连接（cldn5-cldn5、cldn3-cldn3）。Claudin-3和Claudin-5均增强了相互作用强度。这种新方法能够分析单个Claudin对紧密连接完整性的贡献，表征Claudin的同嗜性和异嗜性相互作用。为了测试一种针对屏障效应物的新筛选方法，示例性地，使用这种双细胞卵母细胞模型来分析吸收增强剂癸酸钠对卵母细胞对的影响。
