Pathologic and Phenotypic Alterations in a Mouse Expressing a Connexin47 Missense Mutation That Causes Pelizaeus-Merzbacher–Like Disease in Humans

English version České info

Gap junction channels are intercellular conduits that allow diffusional exchange of ions, second messengers, and metabolites. Human oligodendrocytes express the gap junction protein connexin47 (Cx47), which is encoded by the GJC2 gene. The autosomal recessive mutation hCx47M283T causes Pelizaeus-Merzbacher–like disease 1 (PMLD1), a progressive leukodystrophy characterized by hypomyelination, retarded motor development, nystagmus, and spasticity. We introduced the human missense mutation into the orthologous position of the mouse Gjc2 gene and inserted the mCx47M282T coding sequence into the mouse genome via homologous recombination in embryonic stem cells. Three-week-old homozygous Cx47M282T mice displayed impaired rotarod performance but unchanged open-field behavior. 10-15-day-old homozygous Cx47M282T and Cx47 null mice revealed a more than 80% reduction in the number of cells participating in glial networks after biocytin injections into oligodendrocytes in sections of corpus callosum. Homozygous expression of mCx47M282T resulted in reduced MBP expression and astrogliosis in the cerebellum of ten-day-old mice which could also be detected in Cx47 null mice of the same age. Three-month-old homozygous Cx47M282T mice exhibited neither altered open-field behavior nor impaired rotarod performance anymore. Adult mCx47M282T expressing mice did not show substantial myelin alterations, but homozygous Cx47M282T mice, additionally deprived of connexin32, which is also expressed in oligodendrocytes, died within six weeks after birth and displayed severe myelin defects accompanied by astrogliosis and activated microglia. These results strongly suggest that PMLD1 is caused by the loss of Cx47 channel function that results in impaired panglial coupling in white matter tissue.

Vyšlo v časopise: Pathologic and Phenotypic Alterations in a Mouse Expressing a Connexin47 Missense Mutation That Causes Pelizaeus-Merzbacher–Like Disease in Humans. PLoS Genet 7(7): e32767. doi:10.1371/journal.pgen.1002146
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002146

Souhrn

Zdroje

1. UhlenbergBSchuelkeMRüschendorfFRufNKaindlAM 2004 Mutations in the gene encoding gap junction protein alpha 12 (connexin 46.6) cause Pelizaeus-Merzbacher-like disease. Am J Hum Genet 75 251 260

2. BugianiMAl ShahwanSLamanteaEBizziABakhshE 2006 GJA12 mutations in children with recessive hypomyelinating leukoencephalopathy. Neurology 67 273 279

3. WolfNICundallMRutlandPRosserESurteesR 2007 Frameshift mutation in GJA12 leading to nystagmus, spastic ataxia and CNS dys-/demyelination. Neurogenetics 8 39 44

4. SalviatiLTrevissonEBaldoinMCToldoISartoriS 2007 A novel deletion in the GJA12 gene causes Pelizaeus-Merzbacher-like disease. Neurogenetics 8 57 60

5. HennekeMCombesPDiekmannSBertiniEBrockmannK 2008 GJA12 mutations are a rare cause of Pelizaeus-Merzbacher-like disease. Neurology 70 748 754

6. WangJWangHWangYChenTWuX 2010 Two novel gap junction protein alpha 12 gene mutations in two Chinese patients with Pelizaeus-Merzbacher-like disease. Brain Dev 32 236 243

7. HennekeMGegnerSHahnAPlecko-StartinigBWeschkeB 2010 Clinical neurophysiology in GJA12-related hypomyelination vs Pelizaeus-Merzbacher disease. Neurology 74 1785 1789

8. DiekmannSHennekeMBurckhardtBCGärtnerJ 2010 Pelizaeus-Merzbacher-like disease is caused not only by a loss of connexin47 function but also by a hemichannel dysfunction. Eur J Hum Genet 18 985 992

9. Orthmann-MurphyJLSalsanoEAbramsCKBizziAUzielG 2009 Hereditary spastic paraplegia is a novel phenotype for GJA12/GJC2 mutations. Brain 132 426 438

10. FerrellREBatyCJKimakMAKarlssonJMLawrenceEC 2010 GJC2 missense mutations cause human lymphedema. Am J Hum Genet 86 943 948

11. Orthmann-MurphyJLFreidinMFischerESchererSSAbramsCK 2007 Two distinct heterotypic channels mediate gap junction coupling between astrocyte and oligodendrocyte connexins. J Neurosci 27 13949 13957

12. WilleckeKEibergerJDegenJEckardtDRomualdiA 2002 Structural and functional diversity of connexin genes in the mouse and human genome. Biol Chem 383 725 737

13. OdermattBWellershausKWallraffASeifertGDegenJ 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 4559

14. NagyJIIonescuALynnBDRashJE 2003 Coupling of astrocyte connexins Cx26, Cx30, Cx43 to oligodendrocyte Cx29, Cx32, Cx47: Implications from normal and connexin32 knockout mice. Glia 44 205 218

15. GiaumeCTheisM 2010 Pharmacological and genetic approaches to study connexin-mediated channels in glial cells of the central nervous system. Brain Res Rev 63 160 176

16. AhnMLeeJGustafssonAEnriquezALancasterE 2008 Cx29 and Cx32, two connexins expressed by myelinating glia, do not interact and are functionally distinct. J Neurosci Res 86 992 1006

17. NagyJILiXRempelJStelmackGPatelD 2001 Connexin26 in adult rodent central nervous system: demonstration at astrocytic gap junctions and colocalization with connexin30 and connexin43. J Comp Neurol 441 302 323

18. TheisMJauchRZhuoLSpeidelDWallraffA 2003 Accelerated hippocampal spreading depression and enhanced locomotory activity in mice with astrocyte-directed inactivation of connexin43. J Neurosci 23 766 776

19. FilippovMAHormuzdiSGFuchsECMonyerH 2003 A reporter allele for investigating connexin 26 gene expression in the mouse brain. Eur J Neurosci 18 3183 3192

20. MagnottiLMGoodenoughDAPaulDL 2011 Functional heterotypic interactions between astrocyte and oligodendrocyte connexins. Glia 59 26 34

21. AltevogtBMPaulDL 2004 Four classes of intercellular channels between glial cells in the CNS. J Neurosci 24 4313 4323

22. KamasawaNSikAMoritaMYasumuraTDavidsonKGV 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 86

23. MaglioneMTressOHaasBKarramKTrotterJ 2010 Oligodendrocytes in mouse corpus callosum are coupled via gap junction channels formed by connexin47 and connexin32. Glia 58 1104 1117

24. SargiannidouIVavlitouNAristodemouSHadjisavvasAKyriacouK 2009 Connexin32 mutations cause loss of function in Schwann cells and oligodendrocytes leading to PNS and CNS myelination defects. J Neurosci 29 4736 4749

25. Orthmann-MurphyJLEnriquezADAbramsCKSchererSS 2007 Loss-of-function GJA12/Connexin47 mutations cause Pelizaeus-Merzbacher-like disease. Mol Cell Neurosci 34 629 641

26. MaginTMMcWhirJMeltonDW 1992 A new mouse embryonic stem cell line with good germ line contribution and gene targeting frequency. Nucleic Acids Res 20 3795 3796

27. RodríguezCIBuchholzFGallowayJSequerraRKasperJ 2000 High-efficiency deleter mice show that FLPe is an alternative to Cre-loxP. Nat Genet 25 139 140

28. KleopaKAOrthmannJLEnriquezAPaulDLSchererSS 2004 Unique distributions of the gap junction proteins connexin29, connexin32, and connexin47 in oligodendrocytes. Glia 47 346 357

29. BaumannNPham-DinhD 2001 Biology of oligodendrocyte and myelin in the mammalian central nervous system. Physiol Rev 81 871 927

30. DobrowolskiRSassePSchrickelJWWatkinsMKimJ 2008 The conditional connexin43G138R mouse mutant represents a new model of hereditary oculodentodigital dysplasia in humans. Hum Mol Genet 17 539 554

31. AttalJThéronMCHoudebineLM 1999 The optimal use of IRES (internal ribosome entry site) in expression vectors. Genet Anal 15 161 165

32. SchützMScimemiPMajumderPDe SiatiRDCrispinoG 2010 The human deafness-associated connexin 30 T5M mutation causes mild hearing loss and reduces biochemical coupling among cochlear non-sensory cells in knock-in mice. Hum Mol Genet 19 4759 4773

33. MeusserBHirschCJaroschESommerT 2005 ERAD: the long road to destruction. Nat Cell Biol 7 766 772

34. ParentiRCicirataFZappalàACataniaALa DeliaF 2010 Dynamic expression of Cx47 in mouse brain development and in the cuprizone model of myelin plasticity. Glia 58 1594 1609

35. EliasLABWangDDKriegsteinAR 2007 Gap junction adhesion is necessary for radial migration in the neocortex. Nature 448 901 907

36. DyerCA 2002 The structure and function of myelin: from inert membrane to perfusion pump. Neurochem Res 27 1279 1292

37. TetzlaffW 1982 Tight junction contact events and temporary gap junctions in the sciatic nerve fibres of the chicken during Wallerian degeneration and subsequent regeneration. J Neurocytol 11 839 858

38. WallraffAKöhlingRHeinemannUTheisMWilleckeK 2006 The impact of astrocytic gap junctional coupling on potassium buffering in the hippocampus. J Neurosci 26 5438 5447

39. MenichellaDMMajdanMAwatramaniRGoodenoughDASirkowskiE 2006 Genetic and physiological evidence that oligodendrocyte gap junctions contribute to spatial buffering of potassium released during neuronal activity. J Neurosci 26 10984 10991

40. RashJE 2009 Molecular disruptions of the panglial syncytium block potassium siphoning and axonal saltatory conduction: pertinence to neuromyelitis optica and other demyelinating diseases of the central nervous system. Neuroscience Available at: http://www.ncbi.nlm.nih.gov/pubmed/19850107. Accessed 6 June 2010

41. MatalonRMichalsKSebestaDDeanchingMGashkoffP 1988 Aspartoacylase deficiency and N-acetylaspartic aciduria in patients with Canavan disease. Am J Med Genet 29 463 471

42. MatalonRMichalsKKaulR 1995 Canavan disease: from spongy degeneration to molecular analysis. J Pediatr 127 511 517

43. TrakaMWollmannRLCerdaSRDugasJBarresBA 2008 Nur7 is a nonsense mutation in the mouse aspartoacylase gene that causes spongy degeneration of the CNS. J Neurosci 28 11537 11549

44. SartoriSBurlinaABSalviatiLTrevissonEToldoI 2008 Increased level of N-acetylaspartylglutamate (NAAG) in the CSF of a patient with Pelizaeus-Merzbacher-like disease due to mutation in the GJA12 gene. Eur J Paediatr Neurol 12 348 350

45. MochelFBoildieuNBarritaultJSarretCEymard-PierreE 2010 Elevated CSF N-acetylaspartylglutamate suggests specific molecular diagnostic abnormalities in patients with white matter diseases. Biochim Biophys Acta Available at: http://www.ncbi.nlm.nih.gov/pubmed/20637281. Accessed 29 July 2010

46. BergerUVLuthi-CarterRPassaniLAElkabesSBlackI 1999 Glutamate carboxypeptidase II is expressed by astrocytes in the adult rat nervous system. J Comp Neurol 415 52 64

47. ZollingerMBrauchli-TheotokisJGutteck-AmslerUDoKQStreitP 1994 Release of N-acetylaspartylglutamate from slices of rat cerebellum, striatum, and spinal cord, and the effect of climbing fiber deprivation. J Neurochem 63 1133 1142

48. MoffettJRRossBArunPMadhavaraoCNNamboodiriAMA 2007 N-Acetylaspartate in the CNS: from neurodiagnostics to neurobiology. Prog Neurobiol 81 89 131

49. LiXIonescuAVLynnBDLuSKamasawaN 2004 Connexin47, connexin29 and connexin32 co-expression in oligodendrocytes and Cx47 association with zonula occludens-1 (ZO-1) in mouse brain. Neuroscience 126 611 630

50. LiXPenesMOdermattBWilleckeKNagyJI 2008 Ablation of Cx47 in transgenic mice leads to the loss of MUPP1, ZONAB and multiple connexins at oligodendrocyte-astrocyte gap junctions. Eur J Neurosci 28 1503 1517

51. SourisseauTGeorgiadisATsaparaAAliRRPestellR 2006 Regulation of PCNA and cyclin D1 expression and epithelial morphogenesis by the ZO-1-regulated transcription factor ZONAB/DbpA. Mol Cell Biol 26 2387 2398

52. BaldaMSMatterK 2000 The tight junction protein ZO-1 and an interacting transcription factor regulate ErbB-2 expression. EMBO J 19 2024 2033

53. LimaWRParreiraKSDevuystOCaplanusiAN'kuliF 2010 ZONAB promotes proliferation and represses differentiation of proximal tubule epithelial cells. J Am Soc Nephrol 21 478 488

54. EbnetKSuzukiAOhnoSVestweberD 2004 Junctional adhesion molecules (JAMs): more molecules with dual functions? J Cell Sci 117 19 29

55. AltevogtBMKleopaKAPostmaFRSchererSSPaulDL 2002 Connexin29 is uniquely distributed within myelinating glial cells of the central and peripheral nervous systems. J Neurosci 22 6458 6470

56. MenichellaDMGoodenoughDASirkowskiESchererSSPaulDL 2003 Connexins are critical for normal myelination in the CNS. J Neurosci 23 5963 5973

57. KlugmannMSchwabMHPühlhoferASchneiderAZimmermannF 1997 Assembly of CNS myelin in the absence of proteolipid protein. Neuron 18 59 70

58. PernetVJolySChristFDimouLSchwabME 2008 Nogo-A and myelin-associated glycoprotein differently regulate oligodendrocyte maturation and myelin formation. J Neurosci 28 7435 7444

59. CollinLUsielloAErbsEMathisCBorrelliE 2004 Motor training compensates for cerebellar dysfunctions caused by oligodendrocyte ablation. Proc Natl Acad Sci U S A 101 325 330

60. CrawfordDKMangiardiMXiaXLópez-ValdésHETiwari-WoodruffSK 2009 Functional recovery of callosal axons following demyelination: a critical window. Neuroscience 164 1407 1421

61. EibergerJKibschullMStrenzkeNSchoberABüssowH 2006 Expression pattern and functional characterization of connexin29 in transgenic mice. Glia 53 601 611

62. BuchholzFAngrandPOStewartAF 1996 A simple assay to determine the functionality of Cre or FLP recombination targets in genomic manipulation constructs. Nucleic Acids Res 24 3118 3119

63. TheisMMaginTMPlumAWilleckeK 2000 General or cell type-specific deletion and replacement of connexin-coding DNA in the mouse. Methods 20 205 218

64. DegenJMeierCVan Der GiessenRSSöhlGPetrasch-ParwezE 2004 Expression pattern of lacZ reporter gene representing connexin36 in transgenic mice. J Comp Neurol 473 511 525

65. HaasSBrockhausJVerkhratskyAKettenmannH 1996 ATP-induced membrane currents in ameboid microglia acutely isolated from mouse brain slices. Neuroscience 75 257 261

66. KressinKKuprijanovaEJabsRSeifertGSteinhäuserC 1995 Developmental regulation of Na+ and K+ conductances in glial cells of mouse hippocampal brain slices. Glia 15 173 187

67. TeubnerBOdermattBGuldenagelMSohlGDegenJ 2001 Functional expression of the new gap junction gene connexin47 transcribed in mouse brain and spinal cord neurons. J Neurosci 21 1117 1126

68. TraubOEckertRLichtenberg-FratéHElfgangCBastideB 1994 Immunochemical and electrophysiological characterization of murine connexin40 and −43 in mouse tissues and transfected human cells. Eur J Cell Biol 64 101 112