Abstract
In human glioblastoma, the blood–brain barrier (BBB) is disturbed. According to our concept, the glio-vascular relationships and thus the control of the BBB are essentially dependent on the polarity of astroglial cells. This polarity is characterized by the uneven distribution of the water channel protein aquaporin-4 (AQP4), dystroglycan and other molecules. Recently, we were able to show that the extracellular matrix component agrin is important for the construction and localization of the so-called orthogonal arrays of particles (OAPs), which consist in AQP4. Here, combining freeze-fracture electron microscopy, immunohistochemistry and Western blotting, we describe alterations of expression and distribution of AQP4, dystroglycan, agrin and the matrix metalloproteinases (MMP) 2, 3 and 9 in human primary glioblastomas (eight primary tumours, six recurrent tumours). Increase of MMP3- and MMP2/9 immunoreactivities went along with loss of agrin and dystroglycan respectively. On the protein level, AQP4 expression was increased in glioblastoma compared to control tissue. This was not accompanied by an increase of OAPs, suggesting that AQP4 can also occur without forming OAPs. The results underline our concept of the loss of glioma cell polarity as one of the factors responsible for the disturbance of the neurovascular unit and as an explanation for the formation of edemas in the glioblastoma.
Similar content being viewed by others
References
Agrawal S, Anderson P, Durbeej M, Van Rooijen N, Ivars F, Opdenakker G, Sorokin LM (2006) Dystroglycan is selectively cleaved at the parenchymal basement membrane at sites of leukocyte extravasation in experimental autoimmune encephalomyelitis. J Exp Med 203:1007–1019
Barber AJ, Lieth E (1997) Agrin accumulates in the brain microvascular basal lamina during development of the blood–brain barrier. Dev Dyn 208:62–74
Berzin TM, Zipser BD, Rafii MS, Kuo-Leblanc V, Yancopoulos GD, Glass DJ, Fallon JR, Stopa EG (2000) Agrin and microvascular damage in Alzheimer’s disease. Neurobiol Aging 21:349–355
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Calogero A, Pavoni E, Gramaglia T, D’Amati G, Ragona G, Brancacchioi A, Petrucci TC (2006) Altered expression of α-dystroglycan subunit in human gliomas. Cancer Biol Ther 5:441–448
Candelario-Jalil E, Yang Y, Rosenberg GA (2009) Diverse roles of matrix metalloproteinases and tissue inhibitors of metalloproteinases in neuroinflammation and cerebral ischemia. Neuroscience 158:983–994
Chen F, Ohashi N, Li W, Eckman C, Nguyen JH (2009) Disruptions of occludin and claudin-5 in brain endothelial cells in vitro and in brains of mice with acute liver failure. Hepatology 50:1914–1923
Fallier-Becker P, Sperveslage J, Wolburg H, Noell S (2011) The impact of agrin on the formation of orthogonal arrays of particles in cultured astrocytes from wild-type and agrin-null mice. Brain Res 1367:2–12
Furman CS, Gorelick-Feldman DA, Davidson KG, Yasumura T, Neely JD, Agre P, Rash JE (2003) Aquaporin-4 square array assembly: opposing actions of M1 and M23 isoforms. Proc Natl Acad Sci USA 100:13609–13614
Gidday JM, Gasche YG, Copin J-C, Shah AR, Perez RS, Shapiro SD, Chan PH, Park TS (2005) Leukocyte-derived matrix metalloproteinase-9 mediates blood–brain barrier breakdown and is proinflammatory after transient focal cerebral ischemia. Am J Physiol Heart Circ Physiol 289:H558–H568
Jung JS, Bhat RV, Preston GM, Guggino WB, Baraban JM, Agre P (1994) Molecular characterization of an aquaporin cDNA from brain: candidate osmoreceptor and regulator of water balance. Proc Natl Acad Sci USA 91:13052–13056
Lampert K, Machein U, Machein MR, Conca W, Peter HH, Volk B (1998) Expression of matrix metalloproteinases and their tissue inhibitors in human brain tumours. Am J Pathol 153:429–437
Lee J, Lund-Smith C, Borboa A, Gonzalez AM, Baird A, Eliceiri BP (2009) Glioma-induced remodeling of the neurovascular unit. Brain Res 1288:125–134
Lohmann C, Krischke M, Wegener J, Galla H-J (2004) Tyrosine phosphatase inhibition induces loss of blood–brain barrier integrity by matrix metalloproteinase-dependent and –independent pathways. Brain Res 995:184–196
Mathiisen TM, Lehre KP, Danbolt NC, Ottersen OP (2010) The perivascular astroglial sheath provides a complete covering of the brain microvessels: an electron microscopic 3D reconstruction. Glia 58:1094–1103
McMahan UJ (1990) The agrin hypothesis. Cold Spring Harb Symp Quant Biol 55:407–418
Muir EM, Adcock KH, Morgenstern DA, Clayton R, von Stillfried N, Rhodes K, Ellis C, Fawcett JW, Rogers JH (2002) Matrix metalloproteinases and their inhibitors are produced by overlapping populations of activated astrocytes. Mol Brain Res 100:103–117
Neely JD, Christensen BM, Nielsen S, Agre P (1999) Heterotetrameric composition of aquaporin-4 water channels. Biochemistry 38:11156–11163
Neuhaus J, Schmid E-M, Wolburg H (1990) Stability of orthogonal arrays of particles in murine skeletal muscle and astrocytes after circulatory arrest, and human gliomas. Neurosci Letters 109:163–168
Nico B, Mangieri D, Tamma R, Longo V, Annese T, Crivellato E, Pollo B, Maderna E, Ribatti D, Salmaggi A (2009) Aquaporin-4 contributes to the resolution of peritumoural brain oedema in human glioblastoma multiforme after combined chemotherapy and radiotherapy. Europ J Cancer 45:3315–3325
Noell S, Fallier-Becker P, Beyer C, Kröger S, Mack AF, Wolburg H (2007) Effects of agrin on the expression and distribution of the water channel protein aquaporin-4 and volume regulation in cultured astrocytes. Europ J Neurosci 26:2109–2118
Noell S, Fallier-Becker P, Deutsch U, Mack AF, Wolburg H (2009) Agrin defines polarized distribution of orthogonal arrays of particles in astrocytes. Cell Tissue Res 337:185–195
Noell S, Wolburg-Buchholz K, Mack AF, Beedle AM, Satz JS, Campbell KP, Wolburg H, Fallier-Becker P (2011) Evidence for a role of dystroglycan regulating the membrane architecture of astroglial endfeet. Eur J Neurosci 33:2179–2186
Rascher G, Fischmann A, Kröger S, Duffner F, Grote E-H, Wolburg H (2002) Extracellular matrix and the blood–brain barrier in glioblastoma multiforme: spatial segregation of tenascin and agrin. Acta Neuropathol 104:85–91
Rohlmann A, Gocht A, Wolburg H (1992) Reactive astrocytes in myelin-deficient rat optic nerve reveal an altered distribution of orthogonal arrays of particles (OAP). Glia 5:259–268
Rosell A, Cuadrado E, Ortega-Aznar A, Hernández-Guillamon M, Lo EH, Montaner J (2008) MMP9-positive neutrophil infiltration is associated to blood–brain barrier breakdown and basal lamina type IV collagen degradation during hemorrhagic transformation after human ischemic stroke. Stroke 39:1121–1126
Rosenberg GA (2005) Matrix metalloproteinases and proteolytic opening of the blood–brain barrier in neuroinflammation. In: De Vries E, Prat A (eds) The blood–brain barrier and its microenvironment. Basic physiology to neurological disease. Taylor and Francis, New York, pp 335–358
Rosenberg GA (2009) Matrix metalloproteinases and their multiple roles in neurodegenerative diseases. Lancet Neurol 8:205–216
Solé S, Petegnief V, Gorina R, Chamorro Á, Planas AM (2004) Activation of matrix metalloproteinase-3 and agrin cleavage in cerebral ischemia/reperfusion. J Neuropathol Exp Neurol 63:338–349
Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, Curschmann J, Janzer RC, Ludwin SK, Gorlia T, Allgeier A, Lacombe D, Cairncross JG, Eisenhauer E, Mirimanoff RO, European Organisation for Research and Treatment of Cancer Brain Tumour and Radiotherapy Groups, National Cancer Institute of Canada Clinical Trials Group (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 352:987–996
Stupp R, Hegi ME, Mason WP, van den Bent MJ, Taphoorn MJ, Janzer RC, Ludwin SK, Allgeier A, Fisher B, Belanger K, Hau P, Brandes AA, Gijtenbeek J, Marosi C, Vecht CJ, Mokhtari K, Wesseling P, Villa S, Eisenhauer E, Gorlia T, Weller M, Lacombe D, Cairncross JG, Mirimanoff RO, European Organisation for Research and Treatment of Cancer Brain Tumour and Radiation Oncology Groups, National Cancer Institute of Canada Clinical Trials Group (2009) Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol 10:459–466
Verma S, Kumar M, Gurjav U, Lum S, Nerurkar VR (2010) Reversal of West Nile virus-induced blood–brain barrier disruption and tight junction proteins degradation by matrix metalloproteinases inhibitor. Virology 397:130–138
Warth A, Kröger S, Wolburg H (2004) Redistribution of aquaporin-4 in human glioblastoma correlates with loss of agrin immunoreactivity from brain capillary basal laminae. Acta Neuropathol 107:311–318
Westermark J, Kähäri V-M (1999) Regulation of matrix metalloproteinase expression in tumour invasion. FASEB J 13:781–792
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 105:586–592
Wolburg H, Noell S, Mack A, Wolburg-Buchholz K, Fallier-Becker P (2009a) Brain endothelial cells and the glio-vascular complex. Cell Tissue Res 335:75–96
Wolburg H, Noell S, Wolburg-Buchholz K, Mack AF, Fallier-Becker P (2009b) Agrin, aquaporin-4, and astrocyte polarity as an important feature of the blood–brain barrier. Neuroscientist 15:180–193
Wolburg H, Wolburg-Buchholz K, Fallier-Becker P, Noell S, Mack A (2011) Structure and functions of aquaporin-4-based orthogonal arrays of particles. Int Rev Cell Mol Biol 287:1–41
Wolburg-Buchholz K, Mack AF, Steiner E, Pfeiffer F, Engelhardt B, Wolburg H (2009) Loss of astrocyte polarity marks blood–brain barrier impairment during experimental autoimmune encephalomyelitis. Acta Neuropathologica 118:219–233
Yang Y, Estrada EY, Thompson JF, Liu W, Rosenberg GA (2007) Matrix metalloproteinase-mediated disruption of tight junction proteins in cerebral vessels is reversed by synthetic matrix metalloproteinase inhibitor in focal ischemia in rat. J Cerebr Blood Flow Metab 27:698–709
Zamecnik J (2006) The extracellular space and matrix of gliomas. Acta Neuropathol 110:435–442
Zimmermann DR, Dours-Zimmermann MT (2008) Extracellular matrix of the central nervous system: from neglect to challenge. Histochem Cell Biol 130:635–653
Acknowledgements
Yeliz Donat-Krasnici, Anne Adam, Ria Knittel and Gabi Frommer-Kästle are thanked for their skilful technical assistance. Financial support by the Deutsche Krebshilfe—Mildred Scheel-Stiftung to HW and SN (grants 107686 and 109219) is gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Additional information
This work is dedicated to Prof. Dr. Wolfgang Schlote, the former academic mentor of H.W., on the occasion of his 80th birthday.
Rights and permissions
About this article
Cite this article
Noell, S., Wolburg-Buchholz, K., Mack, A.F. et al. Dynamics of expression patterns of AQP4, dystroglycan, agrin and matrix metalloproteinases in human glioblastoma. Cell Tissue Res 347, 429–441 (2012). https://doi.org/10.1007/s00441-011-1321-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00441-011-1321-4