Abstract
We aimed to examine paracellular barrier function in cultured mouse lung microvascular endothelial cells (LMECs). The transcellular resistance of LMEC monolayers yielded an electrical resistance of approximately 19 Ω × cm2 at days 6 - 7 in culture when the cells reached confluence, and paracellular permeable clearance of sodium fluorescein was the lowest on day 6 in culture, suggesting the formation of tight junctions (TJs) in cultured LMECs. Moreover, the expression of TJ-associated proteins, occludin, claudin-1 and -4 and zonula occludents 1 (ZO-1) was detected in LMECs at day 6 in culture. However, mRNAs of occludin, claudin-1 and -4 and ZO-1 were already expressed on day 1 after culture, and large variations were absent in the mRNA levels of occludin, claudin-4 and ZO-1 between days 1 and 7 in culture, when the level of each mRNA on day 1 in culture was used as a basal level. However, the claudin-1 mRNA level gradually increased up to approximately 7-fold on day 7 in culture over the basal level. These results indicate that the drastic increase in the mRNA expression level of claudin-1 leads to the strong formation of TJs.
Highlights
The lung is a heavily vascularized organ and the gas-blood barrier in the lung consists of single layers ofHow to cite this paper: Ueda, Y., Shinmyouzu, Y., Nakayama, H., Tanino, T., Sakurai, E. and Sakurai, E. (2016) Claudin-1 Leads to Strong Formation of Tight Junction in Cultured Mouse Lung Microvascular Endothelial Cells
Expression of Occludin, Claudin and zonula occludents 1 (ZO-1) Proteins in Mouse lung microvascular endothelial cells (LMECs) The occludin, claudin-1, -4 and ZO-1 bands detected on day 6 in culture were 59, 30, 22 and 139 kDa, respectively, in mouse LMECs (Figure 2). 3.3. mRNA Expression of Occludin, Claudin and ZO-1 Genes The temporal mRNA expression of occludin, claudin and ZO-1 genes in mouse LMECs is shown (Figure 3)
The mRNA of occludin, claudin-1 and -4 and ZO-1 were already expressed on day 1 after culture, and the large variation was absent in the mRNA levels of occludin, claudin-4 and ZO-1 between days 1 and 7 of culture, when the level of each mRNA on day 1 of culture was used as a basal level
Summary
The lung is a heavily vascularized organ and the gas-blood barrier in the lung consists of single layers ofHow to cite this paper: Ueda, Y., Shinmyouzu, Y., Nakayama, H., Tanino, T., Sakurai, E. and Sakurai, E. (2016) Claudin-1 Leads to Strong Formation of Tight Junction in Cultured Mouse Lung Microvascular Endothelial Cells. (2016) Claudin-1 Leads to Strong Formation of Tight Junction in Cultured Mouse Lung Microvascular Endothelial Cells. A regulated paracellular barrier to the movement of water, solutes and immune cells between both epithelial and endothelial cells is created by tight junctions (TJs) [5]. In these cells, TJs are directly involved in barrier and fence functions by sealing them to generate the primary barrier against the diffusion of solutes via the paracellular pathway and by acting as a boundary between the apical and basolateral plasma membrane domains to create and maintain cell polarity [5] [6]. TJ may be closely associated with the actin cytoskeleton in terms of both structure and function [8] [9]
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