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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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References

  • Amado-Azevedo J, Valent ET, Van Nieuw Amerongen GP (2014) Regulation of the endothelial barrier function: a filum granum of cellular forces, Rho-GTPase signaling and microenvironment. Cell Tissue Res 355:557–576

    Article  CAS  PubMed  Google Scholar 

  • Belvitch P, Dudek SM (2012) Role of FAK in S1P-regulated endothelial permeability. Microvasc Res 83:22–30

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • De P, Peng Q, Traktuev DO, Li W, Yoder MC, March KL, Durden DL (2009) Expression of RAC2 in endothelial cells is required for the postnatal neovascular response. Exp Cell Res 315:248–263

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Deramaudt TB, Dujardin D, Hamadi A, Noulet F, Kolli K, De Mey J, Takeda K, Ronde P (2011) FAK phosphorylation at Tyr-925 regulates cross-talk between focal adhesion turnover and cell protrusion. Mol Biol Cell 22:964–975

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Drenckhahn D, Wagner J (1986) Stress fibers in the splenic sinus endothelium in situ: molecular structure, relationship to the extracellular matrix, and contractility. J Cell Biol 102:1738–1747

    Article  CAS  PubMed  Google Scholar 

  • Eliceiri BP, Puente XS, Hood JD, Stupack DG, Schlaepfer DD, Huang XZ, Sheppard D, Cheresh DA (2002) Src-mediated coupling of focal adhesion kinase to integrin alpha(v)beta5 in vascular endothelial growth factor signaling. J Cell Biol 157:149–160

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Goldmann WH (2014) Vinculin-p130Cas interaction is critical for focal adhesion dynamics and mechano-transduction. Cell Biol Int 38:283–286

    Article  CAS  PubMed  Google Scholar 

  • Hirata H, Tatsumi H, Hayakawa K, Sokabe M (2015) Non-channel mechanosensors working at focal adhesion-stress fiber complex. Pflugers Arch 467:141–155

    Article  CAS  PubMed  Google Scholar 

  • Janostiak R, Pataki AC, Brabek J, Rosel D (2014) Mechanosensors in integrin signaling: the emerging role of p130Cas. Eur J Cell Biol 93:445–454

    Article  CAS  PubMed  Google Scholar 

  • Lie PP, Mruk DD, Mok KW, Su L, Lee WM, Cheng CY (2012) Focal adhesion kinase-Tyr407 and -Tyr397 exhibit antagonistic effects on blood-testis barrier dynamics in the rat. Proc Natl Acad Sci U S A 109:12562–12567

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mierke CT (2009) The role of vinculin in the regulation of the mechanical properties of cells. Cell Biochem Biophys 53:115–126

    Article  CAS  PubMed  Google Scholar 

  • Mitra SK, Hanson DA, Schlaepfer DD (2005) Focal adhesion kinase: in command and control of cell motility. Nat Rev Mol Cell Biol 6:56–68

    Article  CAS  PubMed  Google Scholar 

  • Oldenburg J, de Rooij J (2014) Mechanical control of the endothelial barrier. Cell Tissue Res 355:545–555

    Article  CAS  PubMed  Google Scholar 

  • Pai SY, Kim C, Williams DA (2010) Rac GTPases in human diseases. Dis Markers 29:177–187

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Quadri SK (2012) Cross talk between focal adhesion kinase and cadherins: role in regulating endothelial barrier function. Microvasc Res 83:3–11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Roberts GC, Critchley DR (2009) Structural and biophysical properties of the integrin-associated cytoskeletal protein talin. Biophys Rev 1:61–69

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schaller MD (2010) Cellular functions of FAK kinases: insight into molecular mechanisms and novel functions. J Cell Sci 123:1007–1013

    Article  CAS  PubMed  Google Scholar 

  • Shikata Y, Birukov KG, Birukova AA, Verin A, Garcia JG (2003) Involvement of site-specific FAK phosphorylation in sphingosine-1 phosphate- and thrombin-induced focal adhesion remodeling: role of Src and GIT. FASEB J 17:2240–2249

    Article  CAS  PubMed  Google Scholar 

  • Steiniger B, Stachniss V, Schwarzbach H, Barth PJ (2007) Phenotypic differences between red pulp capillary and sinusoidal endothelia help localizing the open splenic circulation in humans. Histochem Cell Biol 128:391–398

    Article  CAS  PubMed  Google Scholar 

  • Su G, Hodnett M, Wu N, Atakilit A, Kosinski C, Godzich M, Huang XZ, Kim JK, Frank JA, Matthay MA, Sheppard D, Pittet JF (2007) Integrin alphavbeta5 regulates lung vascular permeability and pulmonary endothelial barrier function. Am J Respir Cell Mol Biol 36:377–386

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Taguchi K, Ishiuchi T, Takeichi M (2011) Mechanosensitive EPLIN-dependent remodeling of adherens junctions regulates epithelial reshaping. J Cell Biol 194:643–656

    Article  PubMed  PubMed Central  Google Scholar 

  • Tomakidi P, Schulz S, Proksch S, Weber W, Steinberg T (2014) Focal adhesion kinase (FAK) perspectives in mechanobiology: implications for cell behaviour. Cell Tissue Res 357:515–526

    Article  CAS  PubMed  Google Scholar 

  • Uehara K (2006) Distribution of adherens junction mediated by VE-cadherin complex in rat spleen sinus endothelial cells. Cell Tissue Res 323:417–424

    Article  CAS  PubMed  Google Scholar 

  • Uehara K, Miyoshi M (1997) Junctions between the sinus endothelial cells of rat spleen. Cell Tissue Res 287:187–192

    Article  CAS  PubMed  Google Scholar 

  • Uehara K, Miyoshi M (1999) Stress fiber networks in sinus endothelial cells in the rat spleen. Anat Rec 254:22–27

    Article  CAS  PubMed  Google Scholar 

  • Uehara K, Uehara A (2008) Localization of claudin-5 and ZO-1 in rat spleen sinus endothelial cells. Histochem Cell Biol 129:95–103

    Article  CAS  PubMed  Google Scholar 

  • Uehara K, Uehara A (2011) P2Y1, P2Y6, and P2Y12 receptors in rat splenic sinus endothelial cells: an immunohistochemical and ultrastructural study. Histochem Cell Biol 136:557–567

    Article  CAS  PubMed  Google Scholar 

  • Uehara K, Uehara A (2014) Integrin alphavbeta5 in endothelial cells of rat splenic sinus: an immunohistochemical and ultrastructural study. Cell Tissue Res 356:183–193

    Article  CAS  PubMed  Google Scholar 

  • van Nieuw Amerongen GP, Natarajan K, Yin G, Hoefen RJ, Osawa M, Haendeler J, Ridley AJ, Fujiwara K, van Hinsbergh VW, Berk BC (2004) GIT1 mediates thrombin signaling in endothelial cells: role in turnover of RhoA-type focal adhesions. Circ Res 94:1041–1049

    Article  PubMed  Google Scholar 

  • Wan HT, Mruk DD, Li SY, Mok KW, Lee WM, Wong CK, Cheng CY (2013) p-FAK-Tyr(397) regulates spermatid adhesion in the rat testis via its effects on F-actin organization at the ectoplasmic specialization. Am J Physiol Endocrinol Metab 305:E687–E699

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xiao X, Mruk DD, Wong CK, Cheng CY (2014) Germ cell transport across the seminiferous epithelium during spermatogenesis. Physiology (Bethesda) 29:286–298

    CAS  Google Scholar 

  • Yonemura S, Wada Y, Watanabe T, Nagafuchi A, Shibata M (2010) alpha-Catenin as a tension transducer that induces adherens junction development. Nat Cell Biol 12:533–542

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Cell Biology, Fukuoka University School of Medicine, Jonan-ku, Fukuoka, 814-0180, Japan

    Kiyoko Uehara

  2. Department of Physiology, Fukuoka University School of Medicine, Jonan-ku, Fukuoka, 814-0180, Japan

    Akira Uehara

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  1. Kiyoko Uehara
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  2. Akira Uehara
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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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  • DOI: https://doi.org/10.1007/s00441-015-2350-1

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