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Differentiated localizations of phosphorylated focal adhesion kinase in endothelial cells of rat splenic sinus

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Abstract

The splenic sinus endothelium adhering via adherens junctions and tight junctions regulates the passage of blood cells through the splenic cord. Focal adhesion kinase (FAK) regulates the focal adhesion complex in the basal part of endothelial cells and is an integrated component of cell–cell adhesion, depending on its phosphorylation status. The objectives of this study are to assess the localization of FAK phosphorylated at tyrosine residues and the related proteins of integrin β5, talin, paxillin, p130Cas, vinculin, RhoA, Rac1, Rac2, Cdc42 and VE-cadherin, in the sinus endothelial cells of rat spleen and to examine the roles of FAK in regulating endothelial adhesion and the passage of blood cells. Immunofluorescence microscopy of tissue cryosections revealed that FAK was localized in the entire circumference of sinus endothelial cells and FAK phosphorylated at Try397 residue (pFAKy397) and pFAKy576 were precisely localized in the adherens junctions of the endothelial cells, whereas pFAKy925 was localized in the basal part of the endothelial cells. Paxillin and vinculin were prominently localized in the basal part of the endothelial cells. Integrin β5, talin and p130Cas were colocalized with FAK in the entire circumference of sinus endothelial cells. RhoA, Rac2 and Cdc42 were localized in the entire circumference of sinus endothelial cells close to FAK, stress fibers and cortical actin filaments. Immunogold electron microscopy revealed that pFAKy397 and pFAKy576 were colocalized with VE-cadherin, RhoA, Rac2 and Cdc42 in the adherens junctions of the endothelial cells. Possible functional roles of FAK in splenic sinus endothelial cells are also discussed.

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Correspondence to Kiyoko Uehara.

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Uehara, K., Uehara, A. Differentiated localizations of phosphorylated focal adhesion kinase in endothelial cells of rat splenic sinus. Cell Tissue Res 364, 611–622 (2016). https://doi.org/10.1007/s00441-015-2350-1

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Keywords

  • Focal adhesion kinase (FAK)
  • Phosphorylated FAK
  • Adherens junction
  • Endothelial cells
  • Spleen