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01-06-2012 | Original Paper

Gap junction pathology in multiple sclerosis lesions and normal-appearing white matter

Authors: Kyriaki Markoullis, Irene Sargiannidou, Natasa Schiza, Andreas Hadjisavvas, Federico Roncaroli, Richard Reynolds, Kleopas A. Kleopa

Published in: Acta Neuropathologica | Issue 6/2012

Abstract

Oligodendrocyte gap junctions (GJs) are vital for central nervous system myelination, but their involvement in multiple sclerosis (MS) pathology remains unknown. The aim of this study was to examine alterations of oligodendrocyte and related astrocyte GJs in MS lesions and normal-appearing white matter (NAWM). Post-mortem brain samples from 9 MS and 11 age-matched non-MS control patients were studied. Tissue sections that included both chronic active and inactive lesions were characterized neuropathologically with Luxol Fast Blue staining and immunostaining for myelin oligodendrocyte glycoprotein (MOG) and the microglial marker Iba1. We analyzed the expression of Cx32 and Cx47 in oligodendrocytes and of Cx43, the major astrocytic partner in oligodendrocyte–astrocyte (O/A) GJs by quantitative immunoblot and real-time PCR. Formation of GJ plaques was quantified by immunohistochemistry. Compared to control brains, both Cx32 and Cx47 GJ plaques and protein levels were reduced in and around MS lesions, while Cx43 was increased as part of astrogliosis. In the NAWM, Cx32 was significantly reduced along myelinated fibers whereas Cx47 showed increased expression mainly in oligodendrocyte precursor cells (OPCs). However, OPCs showed only limited connectivity to astrocytes. Cx43 showed modestly increased levels in MS NAWM compared to controls, while GJ plaque counts were unchanged. Our findings indicate that oligodendrocyte GJs are affected not only in chronic MS lesions but also in NAWM, where disruption of Cx32 GJs in myelinated fibers may impair myelin structure and function. Moreover, limited O/A GJ connectivity of recruited OPCs in the setting of persistent inflammation and astrogliosis may prevent differentiation and remyelination.

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Ahn M, Lee J, Gustafsson A, Enriquez A, Lancaster E, Sul J, Haydon P, Paul D, Huang Y, Abrams C, Scherer S (2008) Cx29 and Cx32, two connexins expressed by myelinating glia, do not interact and are functionally distinct. J Neurosci Res 86:992–1006PubMedCrossRef

Altevogt BM, Paul DL (2004) Four classes of intercellular channels between glial cells in the CNS. J Neurosci 24:4313–4323PubMedCrossRef

Belliveau DJ, Kidder GM, Vaus CCG (1991) Expression of gap junction genes during postnatal neural development. Dev Genet 12:308–317PubMedCrossRef

Brand-Schieber E, Werner P, Iacobas DA, Iacobas S, Beelitz M, Lowery SL, Spray DC, Scemes E (2005) Connexin43, the major gap junction protein of astrocytes, is down-regulated in inflamed white matter in an animal model of multiple sclerosis. J Neurosci Res 80:798–808PubMedCrossRef

Brück W, Porada P, Poser S, Rieckmann P, Hanefeld F, Kretzschmar HA, Lassmann H (1995) Monocyte/macrophage differentiation in early multiple sclerosis lesions. Ann Neurol 38:788–796PubMedCrossRef

Chang A, Tourtellotte WW, Rudick R, Trapp BD (2002) Premyelinating oligodendrocytes in chronic lesions of multiple sclerosis. N Engl J Med 346:165–173PubMedCrossRef

Ciccarelli O, Werring DJ, Wheeler-Kingshott CA, Barker GJ, Parker GJ, Thompson AJ, Miller DH (2001) Investigation of MS normal-appearing brain using diffusion tensor MRI with clinical correlations. Neurology 56:926–933PubMedCrossRef

Compston A, Coles A (2008) Multiple sclerosis. Lancet 372:1502–1517PubMedCrossRef

Filippi M, Tortorella C, Bozzali M (1999) Normal-appearing white matter changes in multiple sclerosis: the contribution of magnetic resonance techniques. Mult Scler 5:273–282PubMed

10.

Frischer JM, Bramow S, Dal-Bianco A, Lucchinetti CF, Rauschka H, Schmidbauer M, Laursen H, Sorensen PS, Lassmann H (2009) The relation between inflammation and neurodegeneration in multiple sclerosis brains. Brain 132:1175–1189PubMedCrossRef

11.

Frohman EM, Racke MK, Raine CS (2006) Multiple sclerosis—the plaque and its pathogenesis. N Engl J Med 354:942–955PubMedCrossRef

12.

Giaume C, Koulakoff A, Roux L, Holcman D, Rouach N (2010) Astroglial networks: a step further in neuroglial and gliovascular interactions. Nat Rev Neurosci 11:87–99PubMedCrossRef

13.

Hansson E, Muyderman H, Leonova J, Allansson L, Sinclair J, Blomstrand F, Thorlin T, Nilsson M, Ronnback L (2000) Astroglia and glutamate in physiology and pathology: aspects on glutamate transport, glutamate-induced cell swelling and gap-junction communication. Neurochem Int 37:317–329PubMedCrossRef

14.

Howell OW, Rundle JL, Garg A, Komada M, Brophy PJ, Reynolds R (2010) Activated microglia mediate axoglial disruption that contributes to axonal injury in multiple sclerosis. J Neuropathol Exp Neurol 69:1017–1033PubMedCrossRef

15.

Kamasawa N, Sik A, Morita M, Yasumura T, Davidson K, Nagy J, Rash J (2005) Connexin-47 and connexin-32 in gap junctions of oligodendrocyte somata, myelin sheaths, paranodal loops and Schmidt-Lanterman incisures: implications for ionic homeostasis and potassium siphoning. Neuroscience 136:65–86PubMedCrossRef

16.

Kielian T (2008) Glial connexins and gap junctions in CNS inflammation and disease. J Neurochem 106(3):1000–1016PubMedCrossRef

17.

Kleopa KA, Yum SW, Scherer SS (2002) Cellular mechanisms of connexin32 mutations associated with CNS manifestations. J Neurosci Res 68:522–534PubMedCrossRef

18.

Kleopa KA, Orthmann JL, Enriquez A, Paul DL, Scherer SS (2004) Unique distribution of gap junction proteins connexin29, connexin32, and connexin47 in oligodendrocytes. Glia 47:346–357PubMedCrossRef

19.

Kunze A, Congreso MR, Hartmann C, Wallraff-Beck A, Huttmann K, Bedner P, Requardt R, Seifert G, Redecker C, Willecke K, Hoffmann A, Pfeifer A, Theis M, Steinhauser C (2009) Connexin expression by radial glia-like cells is required for neurogenesis in the adult dentate gyrus. Proc Natl Acad Sci USA 106:11336–11341PubMedCrossRef

20.

Kutzelnigg A, Lucchinetti CF, Stadelmann C, Bruck W, Rauschka H, Bergmann M, Schmidbauer M, Parisi JE, Lassmann H (2005) Cortical demyelination and diffuse white matter injury in multiple sclerosis. Brain 128:2705–2712PubMedCrossRef

21.

Lassmann H, Raine CS, Antel J, Prineas JW (1998) Immunopathology of multiple sclerosis: report on an international meeting held at the Institute of Neurology of the University of Vienna. J Neuroimmunol 86:213–217PubMedCrossRef

22.

Lucchinetti CF, Bruck W, Rodriguez M, Lassmann H (1996) Distinct patterns of multiple sclerosis pathology indicates heterogeneity on pathogenesis. Brain Pathol 6:259–274PubMedCrossRef

23.

Lutz SE, Zhao Y, Gulinello M, Lee SC, Raine CS, Brosnan CF (2009) Deletion of astrocyte connexins 43 and 30 leads to a dysmyelinating phenotype and hippocampal CA1 vacuolation. J Neurosci 29:7743–7752PubMedCrossRef

24.

Maglione M, Tress O, Haas B, Karram K, Trotter J, Willecke K, Kettenmann H (2010) Oligodendrocytes in mouse corpus callosum are coupled via gap junction channels formed by connexin47 and connexin32. Glia 58:1104–1117PubMedCrossRef

25.

Magnotti LM, Goodenough DA, Paul DL (2011) Deletion of oligodendrocyte Cx32 and astrocyte Cx43 causes white matter vacuolation, astrocyte loss and early mortality. Glia 59:1064–1074PubMedCrossRef

26.

Melanson-Drapeau L, Beyko S, Dave S, Hebb A, Franks D, Sellitto C, Paul D, Bennett S (2003) Oligodendrocyte progenitor enrichment in the connexin32 null-mutant mouse. J Neurosci 23:1759–1768PubMed

27.

Menichella DM, Goodenough DA, Sirkowski E, Scherer SS, Paul DL (2003) Connexins are critical for normal myelination in the CNS. J Neurosci 23:5963–5973PubMed

28.

Menichella DM, Majdan M, Awatramani R, Goodenough DA, Sirkowski E, Scherer SS, Paul DL (2006) Genetic and physiological evidence that oligodendrocyte gap junctions contribute to spatial buffering of potassium released during neuronal activity. J Neurosci 26(48):10984–10991PubMedCrossRef

29.

Nagy JI, Patel D, Ochalski PAY, Stelmack GL (1999) Connexin30 in rodent, cat and human brain: Selective expression in gray matter astrocytes, co-localization with connexin43 at gap junctions and late developmental appearance. Neuroscience 88(2):447–468PubMedCrossRef

30.

Nagy JI, Rash JE (2000) Connexins and gap junctions of astrocytes and oligodendrocytes in the CNS. Brain Res Rev 32(1):29–44PubMedCrossRef

31.

Nagy JI, Ionescu AV, Lynn BD, Rash JE (2003) Coupling of astrocyte connexins Cx26, Cx30, Cx43 to oligodendrocyte Cx29, Cx32, Cx47: implications from normal and connexin32 knockout mice. Glia 44:205–218PubMedCrossRef

32.

Odermatt B, Wellershaus K, Wallraff A, Seifert G, Degen J, Euwens C, Fuss B, Bussow H, Schilling K, Steinhauser C, Willecke K (2003) Connexin 47 (Cx47)-deficient mice with enhanced green fluorescent protein reporter gene reveal predominant oligodendrocytic expression of Cx47 and display vacuolized myelin in the CNS. J Neurosci 23:4549–4559PubMed

33.

Orthmann-Murphy JL, Enriquez AD, Abrams CK, Scherer SS (2007) Loss-of-function connexin47 mutations cause Pelizaeus–Merzbacher-like disease. Mol Cell Neurosci 34:629–641PubMedCrossRef

34.

Orthmann-Murphy JL, Salsano E, Abrams CK, Bizzi A, Uziel G, Freidin MM, Lamantea E, Zeviani M, Scherer SS, Pareyson D (2009) Hereditary spastic paraplegia is a novel phenotype for GJA12/GJC2 mutations. Brain 132:426–438PubMedCrossRef

35.

Paulson HL, Garbern JY, Hoban TF, Krajewski KM, Lewis RA, Fischbeck KH, Grossman RI, Lenkinski R, Kamholz JA, Shy ME (2002) Transient central nervous system white matter abnormality in X-linked Charcot–Marie–Tooth disease. Ann Neurol 52:429–434PubMedCrossRef

36.

Paznekas WA, Boyadjiev SA, Shapiro R, Daniels O, Wollnik B, Keegan CE, Innis JW, Dinulos MB, Christian C, Hannibal MC, Jabs EW (2003) Connexin 43 (GJA1) mutations cause the pleiotropic phenotype of oculodentodigital dysplasia. Am J Hum Genet 72:408–418PubMedCrossRef

37.

Pelletier D, Nelson SJ, Oh J, Antel JP, Kita M, Zamvil SS, Goodkin DE (2003) MRI lesion volume heterogeneity in primary progressive MS in relation with axonal damage and brain atrophy. J Neurol Neurosurg Psychiatry 74:950–952PubMedCrossRef

38.

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 12:154–169PubMedCrossRef

39.

Prineas JW, Kwon EE, Cho ES, Sharer LR, Barnett MH, Oleszak E, Hoffman B, Morgan BP (2001) Immunopathology of secondary-progressive multiple sclerosis. Ann Neurol 50:646–657PubMedCrossRef

40.

Rash JE, Duffy HS, Dudek FE, Bilhartz BL, Whalen LR, Yasumura T (1997) Grid-mapped freeze-fracture analysis of gap junctions in gray and white matter of adult rat central nervous system, with evidence for a ‘‘panglial syncytium’’ that is not coupled to neurons. J Comp Neurol 388(2):265–292PubMedCrossRef

41.

Rash JE, Yasumura T, Dudek FE, Nagy JI (2001) Cell-specific expression of connexins and evidence of restricted gap junctional coupling between glial cells and between neurons. J Neurosci 21(6):1983–2000PubMed

42.

Retamal MA, Froger N, Palacios-Prado N, Ezan P, Sáez PJ, Sáez JC, Giaume C (2007) Cx43 hemichannels and gap junction channels in astrocytes are regulated oppositely by proinflammatory cytokines released from activated microglia. J Neurosci 27:13781–13792PubMedCrossRef

43.

Reynolds R, Dawson M, Polito A, Cenci di Bello I, Papadopoulos D, Pham-Dinh D, Levine J (2002) The response of NG2-expressing oligodendrocyte progenitors to demyelination in MOG–EAE and MS. J Neurocytol 31:523–536PubMedCrossRef

44.

Reynolds R, Roncaroli F, Nicholas R, Radotra B, Gveric D, Howell O (2011) The neuropathological basis of clinical progression in multiple sclerosis. Acta Neuropathol 122:155–170PubMedCrossRef

45.

Roscoe WA, Messersmith E, Meyer-Franke A, Wipke B, Karlik SJ (2007) Connexin 43 gap junction proteins are up-regulated in remyelinating spinal cord. J Neurosci Res 85:945–953PubMedCrossRef

46.

Sargiannidou I, Ahn M, Enriquez AD, Peinado A, Reynolds R, Abrams CK, Scherer SS, Kleopa KA (2008) Human oligodendrocytes express Cx31.3: function and interactions with Cx32 mutants. Neurobiol Dis 30:221–233PubMedCrossRef

47.

Sargiannidou I, Vavlitou N, Aristodemou S, Hadjisavvas A, Kyriacou K, Scherer SS, Kleopa KA (2009) Connexin32 mutations cause loss of function in Schwann cells and oligodendrocytes leading to PNS and CNS myelination defects. J Neurosci 29:4748–4761CrossRef

48.

Sharma R, Fischer MT, Bauer J, Felts PA, Smith KJ, Misu T, Fujihara K, Bradl M, Lassmann H (2010) Inflammation induced by innate immunity in the central nervous system leads to primary astrocyte dysfunction followed by demyelination. Acta Neuropathol 120:223–236PubMedCrossRef

49.

Soffer D, Raine CS (1980) Morphologic analysis of axo-glial membrane specializations in the demyelinated central nervous system. Brain Res 186:301–313PubMedCrossRef

50.

Sutor B, Schmolke C, Teubner B, Schirmer C, Willecke K (2000) Myelination defects and neuronal hyperexcitability in the neocortex of connexin 32-deficient mice. Cereb Cortex 10(7):684–697PubMedCrossRef

51.

Taylor RA, Simon EM, Marks HG, Scherer SS (2003) The CNS phenotype of X-linked Charcot–Marie–Tooth disease: more than a peripheral problem. Neurology 61:1475–1478PubMedCrossRef

52.

Toews JC, Schram V, Weerth SH, Mignery GA, Russell JT (2007) Signaling proteins in the axoglial apparatus of sciatic nerve nodes of Ranvier. Glia 55(2):202–213PubMedCrossRef

53.

Tress O, Maglione M, Zlomuzica A, May D, Dicke N, Degen J, Dere E, Kettenmann H, Hartmann D, Willecke K (2011) Pathologic and phenotypic alterations in a mouse expressing a connexin47 missense mutation that causes Pelizaeus–Merzbacher-like disease in humans. PLoS Genet 7:e1002146 (Epub 1002011 Jul 1002147)

54.

Uhlenberg B, Schuelke M, Ruschendorf F, Ruf N, Kaindl AM, Henneke M, Thiele H, Stoltenburg-Didinger G, Aksu F, Topaloglu H, Nurnberg P, Hubner C, Weschke B, Gartner J (2004) Mutations in the gene encoding gap junction protein alpha 12 (Connexin 46.6) cause Pelizaeus–Merzbacher-like disease. Am J Hum Genet 75:251–260PubMedCrossRef

55.

Vavlitou N, Sargiannidou I, Markoullis K, Kyriacou K, Scherer SS, Kleopa KA (2010) Axonal pathology precedes demyelination in a mouse model of X-linked demyelinating/type I Charcot-Marie Tooth neuropathy. J Neuropathol Exp Neurol 69:945–958PubMedCrossRef

56.

Véga C, Martiel JL, Drouhault D, Burckhart MF, Coles JA (2003) Uptake of locally applied deoxyglucose, glucose and lactate by axons and Schwann cells of rat vagus nerve. J Physiol 546:551–564PubMedCrossRef

57.

Venance L, Cordier J, Monge M, Zalc B, Glowinski J, Giame C (1995) Homotypic and heterotypic coupling mediated by gap junctions during glial cell differentiation in vitro. Eur J Neurosci 7:451–461PubMedCrossRef

58.

Von Blankenfeld G, Ransom BR, Kettenmann H (1993) Development of cell–cell coupling among cells of the oligodendrocyte lineage. Glia 7:322–328CrossRef

59.

Weiner HL (2009) The Challenge of Multiple Sclerosis: How Do We Cure A Chronic Heterogeneous Disease? Ann Neurol 65:239–248PubMedCrossRef

60.

Wolswijk G (1998) Chronic stage multiple sclerosis lesions contain a relatively quiescent population of oligodendrocyte precursor cells. J Neurosci 18(2):601–609PubMed

61.

Zamvil SS, Steinman L (2003) Diverse targets for intervention during inflammatory and neurodegenerative phases of multiple sclerosis. Neuron 38:685–688PubMedCrossRef

Title: Gap junction pathology in multiple sclerosis lesions and normal-appearing white matter
Authors: Kyriaki Markoullis
Irene Sargiannidou
Natasa Schiza
Andreas Hadjisavvas
Federico Roncaroli
Richard Reynolds
Kleopas A. Kleopa
Publication date: 01-06-2012
Publisher: Springer-Verlag
Published in: Acta Neuropathologica / Issue 6/2012
Print ISSN: 0001-6322
Electronic ISSN: 1432-0533
DOI: https://doi.org/10.1007/s00401-012-0978-4

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Gap junction pathology in multiple sclerosis lesions and normal-appearing white matter

Abstract

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