Uehara Kiyoko, Uehara Akira
Department of Cell Biology, Fukuoka University School of Medicine, Jonan-ku, Fukuoka, 814-0180, Japan.
Histochem Cell Biol. 2008 Jan;129(1):95-103. doi: 10.1007/s00418-007-0343-1. Epub 2007 Sep 25.
Splenic sinus endothelial cells, which adhere through tight and adherens junctions, regulate the passage of blood cells through the splenic cord. The objective of this study was to assess the localization of tight junctional proteins, claudin-5 and ZO-1 in the sinus endothelial cells of rat spleen and to characterize spatial and functional relationships between tight and adherens junctions. Immunofluorescence microscopy of tissue cryosections demonstrated that claudin-5, ZO-1, and alpha-catenin were distinctly localized in the junctional regions of adjacent endothelial cells. Immunogold electron microscopy demonstrated claudin-5 localized in the tight-junctional fused membranes of adjacent endothelial cells. Immunogold labeling for ZO-1 was localized not only in the tight-junctional-fused membranes of endothelial cells but also in the junctional membrane. alpha-Catenin was intermittently localized along the juxtaposed junctional membranes of adjacent endothelial cells. Double-staining immunogold microscopy for claudin-5 and ZO-1, claudin-5 and VE-cadherin, ZO-1 and VE-cadherin, and ZO-1 and alpha-catenin demonstrated that ZO-1 was closely localized to VE-cadherin and alpha-catenin in their juxtaposed membranes of endothelial cells. Thus, ZO-1 might play an important role in regulating the cell-cell junctions of sinus endothelial cells for blood-cell passage through splenic cords.
脾窦内皮细胞通过紧密连接和黏附连接相互黏附,调节血细胞通过脾索的过程。本研究的目的是评估紧密连接蛋白claudin-5和ZO-1在大鼠脾脏窦内皮细胞中的定位,并表征紧密连接和黏附连接之间的空间和功能关系。组织冰冻切片的免疫荧光显微镜检查表明,claudin-5、ZO-1和α-连环蛋白明显定位于相邻内皮细胞的连接区域。免疫金电子显微镜显示claudin-5定位于相邻内皮细胞的紧密连接融合膜中。ZO-1的免疫金标记不仅定位于内皮细胞的紧密连接融合膜中,也定位于连接膜中。α-连环蛋白沿相邻内皮细胞并列的连接膜间歇性定位。对claudin-5和ZO-1、claudin-5和VE-钙黏蛋白、ZO-1和VE-钙黏蛋白以及ZO-1和α-连环蛋白进行双重染色免疫金显微镜检查表明,ZO-1在其内皮细胞并列膜中与VE-钙黏蛋白和α-连环蛋白紧密定位。因此,ZO-1可能在调节窦内皮细胞的细胞间连接以促进血细胞通过脾索方面发挥重要作用。