Advertisement

Acta Neuropathologica

, Volume 114, Issue 5, pp 459–469 | Cite as

Increased caveolin-1 expression precedes decreased expression of occludin and claudin-5 during blood–brain barrier breakdown

  • Sukriti NagEmail author
  • Roopa Venugopalan
  • Duncan J. Stewart
Original Paper

Abstract

The significance of caveolin-1, a major constituent of caveolae, and the tight junction proteins occludin and claudin-5 in early blood–brain barrier (BBB) breakdown was assessed by sequential demonstration of the expression of these proteins over a period of 12 h to 6 days post-lesion in the rat cortical cold injury model. Pial and intracerebral vessels of control rats showed punctuate endothelial immunoreactivity for caveolin-1 and caveolin-2, while claudin-5 and occludin were localized as longitudinal strands in endothelium. During the early phase of BBB breakdown following injury at 12 h and on day 2, western blot analyses detected a significant increase in caveolin-1 expression at the lesion site while immunohistochemistry showed that the caveolin-1 increase was localized to the endothelium of lesion vessels. Decreased expression of occludin occurred at the lesion site only on days 2 and 4 post-lesion while claudin-5 expression was decreased only on day 2. Dual labeling for fibronectin, a marker of BBB breakdown, and caveolin-1 or the tight junction proteins demonstrated that only lesion vessels with BBB breakdown showed a marked increase of caveolin-1, loss of occludin and reduced localization of claudin-5. The issue whether these alterations precede or follow BBB breakdown is uncertain; however, increased expression of caveolin-1 preceded the decreased expression of occludin and claudin-5. Thus caveolae and caveolin-1 have an important role in early BBB breakdown and could be potential therapeutic targets in the control of early brain edema.

Keywords

Blood–brain barrier Caveolae Caveolin-1 Claudin-5 Occludin Cold-injury Tight junctions 

Notes

Acknowledgments

The authors thank Dr. James Eubanks for helpful suggestions throughout this work and for reviewing this manuscript. Supported by Heart and Stroke Foundation of Ontario, Grant #5347

References

  1. 1.
    Ahn M, Kim H, Kim JT, Lee J, Hyun JW, Park JW, Shin T (2006) Gamma-ray irradiation stimulates the expression of caveolin-1 and GFAP in rat spinal cord: a study of immunoblot and immunohistochemistry. J Vet Sci 7(4):309–314PubMedGoogle Scholar
  2. 2.
    Anderson RG, Kamen BA, Rothberg KG, Lacey SW (1992) Potocytosis: sequestration and transport of small molecules by caveolae. Science 255:410–411PubMedCrossRefGoogle Scholar
  3. 3.
    Ballabh P, Hu F, Kumarasiri M, Braun A, Nedergaard M (2005) Development of tight junction molecules in blood vessels of germinal matrix, cerebral cortex, and white matter. Pediatr Res 58:791–798PubMedCrossRefGoogle Scholar
  4. 4.
    Bolton SJ, Anthony DC, Perry VH (1998) Loss of the tight junction proteins occludin and zonula occludens-1 from cerebral vascular endothelium during neutrophil-induced blood–brain barrier breakdown in vivo. Neuroscience 86:1245–1257PubMedCrossRefGoogle Scholar
  5. 5.
    Boyd NL, Park H, Yi H, Boo YC, Sorescu GP, Sykes M, Jo H (2003) Chronic shear induces caveolae formation and alters ERK and Akt responses in endothelial cells. Am J Physiol Heart Circ Physiol 285:H1113–H1122PubMedGoogle Scholar
  6. 6.
    Dallasta LM, Pisarov LA, Esplen JE, Werley JV, Moses AV, Nelson JA, Achim CL (1999) Blood–brain barrier tight junction disruption in human immunodeficiency virus-1 encephalitis. Am J Pathol 155:1915–1927PubMedGoogle Scholar
  7. 7.
    Furuse M, Sasaki H, Fujimoto K, Tsukita S (1998) A single gene product, claudin-1 or -2, reconstitutes tight junction strands and recruits occludin in fibroblasts. J Cell Biol 143:391–401PubMedCrossRefGoogle Scholar
  8. 8.
    Gow A, Southwood CM, Li JS, Pariali M, Riordan GP, Brodie SE, Danias J, Bronstein JM, Kachar B, Lazzarini RA (1999) CNS myelin and sertoli cell tight junction strands are absent in Osp/claudin-11 null mice. Cell 99:649–659PubMedCrossRefGoogle Scholar
  9. 9.
    Hirase T, Staddon JM, Saitou M, Ando-Akatsuka Y, Itoh M, Furuse M, Fujimoto K, Tsukita S, Rubin LL (1997) Occludin as a possible determinant of tight junction permeability in endothelial cells. J Cell Sci 110:1603–1613PubMedGoogle Scholar
  10. 10.
    Huber JD, Egleton RD, Davis TP (2001) Molecular physiology and pathophysiology of tight junctions in the blood–brain barrier. Trends Neurosci 24:719–725PubMedCrossRefGoogle Scholar
  11. 11.
    Ikezu T, Ueda H, Trapp BD, Nishiyama K, Sha JF, Volonte D, Galbiati F, Byrd AL, Bassell G, Serizawa H, Lane WS, Lisanti MP, Okamoto T (1998) Affinity-purification and characterization of caveolins from the brain: differential expression of caveolin-1, -2, and -3 in brain endothelial and astroglial cell types. Brain Res 804:177–192PubMedCrossRefGoogle Scholar
  12. 12.
    Kubota K, Furuse M, Sasaki H, Sonoda N, Fujita K, Nagafuchi A, Tsukita S (1999) Ca(2+)-independent cell-adhesion activity of claudins, a family of integral membrane proteins localized at tight junctions. Curr Biol 9:1035–1038PubMedCrossRefGoogle Scholar
  13. 13.
    Liebner S, Fischmann A, Rascher G, Duffner F, Grote EH, Kalbacher H, Wolburg H (2000) Claudin-1 and claudin-5 expression and tight junction morphology are altered in blood vessels of human glioblastoma multiforme. Acta Neuropathol 100:323–331PubMedCrossRefGoogle Scholar
  14. 14.
    Liebner S, Kniesel U, Kalbacher H, Wolburg H (2000) Correlation of tight junction morphology with the expression of tight junction proteins in blood–brain barrier endothelial cells. Eur J Cell Biol 79:707–717PubMedCrossRefGoogle Scholar
  15. 15.
    Lipardi C, Mora R, Colomer V, Paladino S, Nitsch L, Rodriguez-Boulan E, Zurzolo C (1998) Caveolin transfection results in caveolae formation but not apical sorting of glycosylphosphatidylinositol (GPI)-anchored proteins in epithelial cells. J Cell Biol 140:617–626PubMedCrossRefGoogle Scholar
  16. 16.
    Lossinsky AS, Shivers RR (2004) Structural pathways for macromolecular and cellular transport across the blood–brain barrier during inflammatory conditions. Rev Histol Histopathol 19:535–564Google Scholar
  17. 17.
    Mayhan WG (2000) Nitric oxide donor-induced increase in permeability of the blood–brain barrier. Brain Res 866:101–108PubMedCrossRefGoogle Scholar
  18. 18.
    McCarthy KM, Francis SA, McCormack JM, Lai J, Rogers RA, Skare IB, Lynch RD, Schneeberger EE (2000) Inducible expression of claudin-1-myc but not occludin-VSV-G results in aberrant tight junction strand formation in MDCK cells. J Cell Sci 113:3387–3398PubMedGoogle Scholar
  19. 19.
    Monier S, Parton RG, Vogel F, Behlke J, Henske A, Kurzchalia TV (1995) VIP21-caveolin, a membrane protein constituent of the caveolar coat, oligomerizes in vivo and in vitro. Mol Biol Cell 6:911–927PubMedGoogle Scholar
  20. 20.
    Morita K, Sasaki H, Furuse M, Tsukita S (1999) Endothelial claudin: Claudin-5/TMVCF constitutes tight junction strands in endothelial cells. J Cell Biol 147:185–194PubMedCrossRefGoogle Scholar
  21. 21.
    Nag S (1998) Blood–brain barrier permeability measured with histochemistry. In: Pardridge WM (ed) Introduction to the blood–brain barrier. Methodology, biology and pathology. Cambridge University Press, Cambridge, pp 113–121Google Scholar
  22. 22.
    Nag S (2003) Pathophysiology of blood–brain barrier breakdown. Methods Mol Med 89:97–119PubMedGoogle Scholar
  23. 23.
    Nag S (2005) Anatomy and structure of brain blood vessels. In: Kalimo H (ed) Pathology and genetics. Cerebrovascular diseases. ISN Neuropath Press, Basel, pp 14–21Google Scholar
  24. 24.
    Nag S, Eskandarian MR, Davis J, Eubanks JH (2002) Differential expression of vascular endothelial growth factor-A (VEGF-A) and VEGF-B after brain injury. J Neuropathol Exp Neurol 61:778–788PubMedGoogle Scholar
  25. 25.
    Nag S, Papneja T, Venugopalan R, Stewart DJ (2005) Increased angiopoietin2 expression is associated with endothelial apoptosis and blood–brain barrier breakdown. Lab Invest 85:1189–1198PubMedCrossRefGoogle Scholar
  26. 26.
    Nag S, Picard P, Stewart DJ (2001) Expression of nitric oxide synthases and nitrotyrosine during blood–brain barrier breakdown and repair after cold injury. Lab Invest 81:41–49PubMedGoogle Scholar
  27. 27.
    Nag S, Takahashi JL, Kilty DW (1997) Role of vascular endothelial growth factor in blood–brain barrier breakdown and angiogenesis in brain trauma. J Neuropathol Exp Neurol 56:912–921PubMedCrossRefGoogle Scholar
  28. 28.
    Nitta T, Hata M, Gotoh S, Seo Y, Sasaki H, Hashimoto N, Furuse M, Tsukita S (2003) Size-selective loosening of the blood–brain barrier in claudin-5-deficient mice. J Cell Biol 161:653–660PubMedCrossRefGoogle Scholar
  29. 29.
    Nourhaghighi N, Teichert-Kuliszewska K, Davis J, Stewart DJ, Nag S (2003) Altered expression of angiopoietins during blood–brain barrier breakdown and angiogenesis. Lab Invest 83:1211–1222PubMedCrossRefGoogle Scholar
  30. 30.
    Parton RG, Hanzal-Bayer M, Hancock JF (2006) Biogenesis of caveolae: a structural model for caveolin-induced domain formation. J Cell Sci 119:787–796PubMedCrossRefGoogle Scholar
  31. 31.
    Parton RG, Simons K (2007) The multiple faces of caveolae. Nat Rev Mol Cell Biol 8:185–194PubMedCrossRefGoogle Scholar
  32. 32.
    Plumb J, McQuaid S, Mirakhur M, Kirk J (2002) Abnormal endothelial tight junctions in active lesions and normal-appearing white matter in multiple sclerosis. Brain Pathol 12(2):154–169PubMedCrossRefGoogle Scholar
  33. 33.
    Predescu D, Vogel SM, Malik AB (2004) Functional and morphological studies of protein transcytosis in continuous endothelia. Am J Physiol Lung Cell Mol Physiol 287:L895–L901PubMedCrossRefGoogle Scholar
  34. 34.
    Reese TS, Karnovsky MJ (1967) Fine structural localization of a blood–brain barrier to exogenous peroxidase. J Cell Biol 34:207–217PubMedCrossRefGoogle Scholar
  35. 35.
    Rizzo V, Morton C, DePaola N, Schnitzer JE, Davies PF (2003) Recruitment of endothelial caveolae into mechanotransduction pathways by flow conditioning in vitro. Am J Physiol Heart Circ Physiol 285:H1720–H1729PubMedGoogle Scholar
  36. 36.
    Saitou M, Furuse M, Sasaki H, Schulzke JD, Fromm M, Takano H, Noda T, Tsukita S (2000) Complex phenotype of mice lacking occludin, a component of tight junction strands. Mol Biol Cell 11:4131–4142PubMedGoogle Scholar
  37. 37.
    Scherer PE, Okamoto T, Chun M, Nishimoto I, Lodish HF, Lisanti MP (1996) Identification, sequence, and expression of caveolin-2 defines a caveolin gene family. Proc Natl Acad Sci USA 93:131–135PubMedCrossRefGoogle Scholar
  38. 38.
    Schnitzer JE, Oh P, McIntosh DP (1996) Role of GTP hydrolysis in fission of caveolae directly from plasma membranes. Science 274:239–242PubMedCrossRefGoogle Scholar
  39. 39.
    Schubert W, Frank PG, Razani B, Park DS, Chow CW, Lisanti MP (2001) Caveolae-deficient endothelial cells show defects in the uptake and transport of albumin in vivo. J Biol Chem 276:48619–48622PubMedCrossRefGoogle Scholar
  40. 40.
    Shin T, Kim H, Jin JK, Moon C, Ahn M, Tanuma N, Matsumoto Y (2005) Expression of caveolin-1, -2, and -3 in the spinal cords of Lewis rats with experimental autoimmune encephalomyelitis. J Neuroimmunol 165:11–20PubMedCrossRefGoogle Scholar
  41. 41.
    Song L, Ge S, Pachter JS (2007) Caveolin-1 regulates expression of junction-associated proteins in brain microvascular endothelial cells. Blood 109:1515–1523PubMedCrossRefGoogle Scholar
  42. 42.
    Tsukita S, Furuse M (1999) Occludin and claudins in tight-junction strands: leading or supporting players? Tr Cell Biol 9:268–273CrossRefGoogle Scholar
  43. 43.
    Unterberg A, Wahl M, Baethmann A (1984) Effects of bradykinin on permeability and diameter of pial vessels in vivo. J Cereb Blood Flow Metab 4:574–585PubMedGoogle Scholar
  44. 44.
    Van Itallie CM, Anderson JM (2006) Claudins and epithelial paracellular transport. Annu Rev Physiol 68:403–429PubMedCrossRefGoogle Scholar
  45. 45.
    Virgintino D, Errede M, Robertson D, Capobianco C, Girolamo F, Vimercati A, Bertossi M, Roncali L (2004) Immunolocalization of tight junction proteins in the adult and developing human brain. Histochem Cell Biol 122:51–59PubMedCrossRefGoogle Scholar
  46. 46.
    Virgintino D, Robertson D, Errede M, Benagiano V, Tauer U, Roncali L, Bertossi M (2002) Expression of caveolin-1 in human brain microvessels. Neuroscience 115:145–152PubMedCrossRefGoogle Scholar
  47. 47.
    Witt KA, Mark KS, Hom S, Davis TP (2003) Effects of hypoxia-reoxygenation on rat blood–brain barrier permeability and tight junctional protein expression. Am J Physiol Heart Circ Physiol 285:H2820–H2831PubMedGoogle Scholar
  48. 48.
    Wolburg H, Wolburg-Buchholz K, Kraus J, Rascher-Eggstein G, Liebner S, Hamm S, Duffner F, Grote EH, Risau W, Engelhardt B (2003) Localization of claudin-3 in tight junctions of the blood–brain barrier is selectively lost during experimental autoimmune encephalomyelitis and human glioblastoma multiforme. Acta Neuropathol (Berl) 105:586–592Google Scholar
  49. 49.
    Woodman SE, Ashton AW, Schubert W, Lee H, Williams TM, Medina FA, Wyckoff JB, Combs TP, Lisanti MP (2003) Caveolin-1 knockout mice show an impaired angiogenic response to exogenous stimuli. Am J Pathol 162:2059–2068PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Sukriti Nag
    • 1
    • 3
    Email author
  • Roopa Venugopalan
    • 1
  • Duncan J. Stewart
    • 2
  1. 1.Toronto Western Research Institute, University Health NetworkUniversity of TorontoTorontoCanada
  2. 2.Terrance Donnelly Heart Center, St Michael’s HospitalUniversity of TorontoTorontoCanada
  3. 3.Department of Laboratory Medicine and PathobiologyBanting InstituteTorontoCanada

Personalised recommendations