Journal of Molecular Neuroscience

, Volume 41, Issue 1, pp 172–182 | Cite as

SAGE Analysis of Genes Differentially Expressed in Presymptomatic TgSOD1G93A Transgenic Mice Identified Cellular Processes Involved in Early Stage of ALS Pathology

  • Michel Guipponi
  • Qiao-Xin Li
  • Lavinia Hyde
  • Tim Beissbarth
  • Gordon K. Smyth
  • Colin L. Masters
  • Hamish S. Scott
Article

Abstract

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative condition in which motor neurons of the spinal cord and motor cortex degenerate, resulting in progressive paralysis. Transgenic mice expressing human mutant Cu/Zn superoxide dismutase-1 (SOD1) present a pathology that is very similar to that seen in human ALS patients. Using serial analysis of gene expression, we investigated the effects of mutant human SOD1 protein on global gene expression in the spinal cord and lower brain stem of presymptomatic TgSOD1G93A transgenic mice. One hundred twenty transcripts were found to be significantly dysregulated in the presence of mutant SOD1 protein, 79 being down-regulated and 41 up-regulated. Quantitative RT-PCR was used to confirm the differential expression of nine of these genes. Immunohistochemistry analysis on spinal cord sections revealed that dysregulation of these mutant SOD1-induced molecular pathways are concomitant to the appearance of discrete signs of neuropathology including neuronal loss, elevated gliosis, and ubiquitin-positive deposits. Altogether, our data showed that early signs of neuropathology in the SOD1 mutant mice are accompanied by altered expression of genes involved in various biological processes including apoptosis, oxidative stress, ATP biosynthesis, myelination, and axonal transport.

Keywords

Amyotrophic lateral sclerosis SOD1G93A Serial analysis of gene expression (SAGE) Transgenic mice Spinal cord 

References

  1. Bendotti C, Atzori C, Piva R, Tortarolo M, Strong MJ, DeBiasi S, Migheli A (2004) Activated p38MAPK is a novel component of the intracellular inclusions found in human amyotrophic lateral sclerosis and mutant SOD1 transgenic mice. J Neuropathol Exp Neurol 63:113–119PubMedGoogle Scholar
  2. Benjamini Y, Drai D, Elmer G, Kafkafi N, Golani I (2001) Controlling the false discovery rate in behavior genetics research. Behav Brain Res 125:279–284CrossRefPubMedGoogle Scholar
  3. Boillee S, Vande Velde C, Cleveland D (2006) ALS: a disease of motor neurons and their nonneuronal neighbors. Neuron 52:39–59CrossRefPubMedGoogle Scholar
  4. Boyd-Kimball D, Castegna A, Sultana R, Poon HF, Petroze R, Lynn BC, Klein JB, Butterfield DA (2005) Proteomic identification of proteins oxidized by Abeta(1–42) in synaptosomes: implications for Alzheimer's disease. Brain Res 1044:206–215CrossRefPubMedGoogle Scholar
  5. Bruijn LI, Miller TM, Cleveland DW (2004) Unraveling the mechanisms involved in motor neuron degeneration in ALS. Annu Rev Neurosci 27:723–749CrossRefPubMedGoogle Scholar
  6. Casoni F, Basso M, Massignan T, Gianazza E, Cheroni C, Salmona M, Bendotti C, Bonetto V (2005) Protein nitration in a mouse model of familial amyotrophic lateral sclerosis: possible multifunctional role in the pathogenesis. J Biol Chem 280:16295–16304CrossRefPubMedGoogle Scholar
  7. Danial NN, Gramm CF, Scorrano L et al (2003) BAD and glucokinase reside in a mitochondrial complex that integrates glycolysis and apoptosis. Nature 424:952–956CrossRefPubMedGoogle Scholar
  8. Di Giorgio FP, Carrasco MA, Siao MC, Maniatis T, Eggan K (2007) Non-cell autonomous effect of glia on motor neurons in an embryonic stem cell-based ALS model. Nat Neurosci 10:608–614CrossRefPubMedGoogle Scholar
  9. Ferraiuolo L, Heath PR, Holden H, Kasher P, Kirby J, Shaw PJ (2007) Microarray analysis of the cellular pathways involved in the adaptation to and progression of motor neuron injury in the SOD1 G93A mouse model of familial ALS. J Neurosci 27:9201–9219CrossRefPubMedGoogle Scholar
  10. Gitcho MA, Baloh RH, Chakraverty S et al (2008) TDP-43 A315T mutation in familial motor neuron disease. Ann Neurol 63:535–538CrossRefPubMedGoogle Scholar
  11. Gonin-Giraud S, Mathieu AL, Diocou S, Tomkowiak M, Delorme G, Marvel J (2002) Decreased glycolytic metabolism contributes to but is not the inducer of apoptosis following IL-3-starvation. Cell Death Differ 9:1147–1157CrossRefPubMedGoogle Scholar
  12. Gurney ME, Pu H, Chiu AY et al (1994) Motor neuron degeneration in mice that express a human Cu, Zn superoxide dismutase mutation. Science 264:1772–1775CrossRefPubMedGoogle Scholar
  13. Hatfield MD, Reis AM, Obeso D, Cook JR, Thompson DM, Rao M, Friedberg EC, Queimado L (2006) Identification of MMS19 domains with distinct functions in NER and transcription. DNA Repair (Amst) 5:914–924CrossRefGoogle Scholar
  14. Hiraiwa M, Campana WM, Mizisin AP, Mohiuddin L, O'Brien JS (1999) Prosaposin: a myelinotrophic protein that promotes expression of myelin constituents and is secreted after nerve injury. Glia 26:353–360CrossRefPubMedGoogle Scholar
  15. Klivenyi P, Ferrante RJ, Matthews RT et al (1999) Neuroprotective effects of creatine in a transgenic animal model of amyotrophic lateral sclerosis. Nat Med 5:347–350CrossRefPubMedGoogle Scholar
  16. Kwiatkowski TJ Jr, Bosco DA, Leclerc AL et al (2009) Mutations in the FUS/TLS gene on chromosome 16 cause familial amyotrophic lateral sclerosis. Science 323:1205–1208CrossRefPubMedGoogle Scholar
  17. Lee YJ, Keng PC (2005) Studying the effects of actin cytoskeletal destabilization on cell cycle by cofilin overexpression. Mol Biotechnol 31:1–10CrossRefPubMedGoogle Scholar
  18. Li QX, Mok SS, Laughton KM, McLean CA, Volitakis I, Cherny RA, Cheung NS, White AR, Masters CL (2006) Overexpression of Abeta is associated with acceleration of onset of motor impairment and superoxide dismutase 1 aggregation in an amyotrophic lateral sclerosis mouse model. Aging Cell 5:153–165CrossRefPubMedGoogle Scholar
  19. Liu J, Lillo C, Jonsson PA et al (2004) Toxicity of familial ALS-linked SOD1 mutants from selective recruitment to spinal mitochondria. Neuron 43:5–17CrossRefPubMedGoogle Scholar
  20. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  21. Lukas TJ, Luo WW, Mao H, Cole N, Siddique T (2006) Informatics-assisted protein profiling in a transgenic mouse model of amyotrophic lateral sclerosis. Mol Cell Proteomics 5:1233–1244CrossRefPubMedGoogle Scholar
  22. Manfredi G, Xu Z (2005) Mitochondrial dysfunction and its role in motor neuron degeneration in ALS. Mitochondrion 5:77–87CrossRefPubMedGoogle Scholar
  23. Maraganore DM, Lesnick TG, Elbaz A et al (2004) UCHL1 is a Parkinson's disease susceptibility gene. Ann Neurol 55:512–521CrossRefPubMedGoogle Scholar
  24. Munch C, Sedlmeier R, Meyer T et al (2004) Point mutations of the p150 subunit of dynactin (DCTN1) gene in ALS. Neurology 63:724–726PubMedGoogle Scholar
  25. Nagai M, Re DB, Nagata T, Chalazonitis A, Jessell TM, Wichterle H, Przedborski S (2007) Astrocytes expressing ALS-linked mutated SOD1 release factors selectively toxic to motor neurons. Nat Neurosci 10:615–622CrossRefPubMedGoogle Scholar
  26. Newbery HJ, Gillingwater TH, Dharmasaroja P, Peters J, Wharton SB, Thomson D, Ribchester RR, Abbott CM (2005) Progressive loss of motor neuron function in wasted mice: effects of a spontaneous null mutation in the gene for the eEF1 A2 translation factor. J Neuropathol Exp Neurol 64:295–303PubMedGoogle Scholar
  27. Offen D, Barhum Y, Melamed E, Embacher N, Schindler C, Ransmayr G (2009) Spinal cord mRNA profile in patients with ALS: Comparison with transgenic mice expressing the human SOD-1 mutant. J Mol Neurosci 38:85–93CrossRefPubMedGoogle Scholar
  28. Park JH, Hong YH, Kim HJ, Kim SM, Kim MJ, Park KS, Sung JJ, Lee KW (2007) Pyruvate slows disease progression in a G93A SOD1 mutant transgenic mouse model. Neurosci Lett 413:265–269CrossRefPubMedGoogle Scholar
  29. Pasinelli P, Belford ME, Lennon N, Bacskai BJ, Hyman BT, Trotti D, Brown RH Jr (2004) Amyotrophic lateral sclerosis-associated SOD1 mutant proteins bind and aggregate with Bcl-2 in spinal cord mitochondria. Neuron 43:19–30CrossRefPubMedGoogle Scholar
  30. Perluigi M, Fai PH, Hensley K, Pierce WM, Klein JB, Calabrese V, De Marco C, Butterfield DA (2005) Proteomic analysis of 4-hydroxy-2-nonenal-modified proteins in G93A-SOD1 transgenic mice-A model of familial amyotrophic lateral sclerosis. Free Radic Biol Med 38:960–968CrossRefPubMedGoogle Scholar
  31. Perrin FE, Boisset G, Docquier M, Schaad O, Descombes P, Kato AC (2005) No widespread induction of cell death genes occurs in pure motoneurons in an amyotrophic lateral sclerosis mouse model. Hum Mol Genet 14:3309–3320CrossRefPubMedGoogle Scholar
  32. Ran Q, Liang H, Gu M, Qi W, Walter CA, Roberts LJ II, Herman B, Richardson A, Van Remmen H (2004) Transgenic mice overexpressing glutathione peroxidase 4 are protected against oxidative stress-induced apoptosis. J Biol Chem 279:55137–55146CrossRefPubMedGoogle Scholar
  33. Rutherford NJ, Zhang YJ, Baker M et al (2008) Novel mutations in TARDBP (TDP-43) in patients with familial amyotrophic lateral sclerosis. PLoS Genet 4:e1000193CrossRefPubMedGoogle Scholar
  34. Saha S, Sparks AB, Rago C, Akmaev V, Wang CJ, Vogelstein B, Kinzler KW, Velculescu VE (2002) Using the transcriptome to annotate the genome. Nat Biotechnol 20:508–512CrossRefPubMedGoogle Scholar
  35. Sasaki S, Warita H, Abe K, Iwata M (2004) Slow component of axonal transport is impaired in the proximal axon of transgenic mice with a G93A mutant SOD1 gene. Acta Neuropathol 107:452–460CrossRefPubMedGoogle Scholar
  36. Sau D, De Biasi S, Vitellaro-Zuccarello L et al (2007) Mutation of SOD1 in ALS: a gain of a loss of function. Hum Mol Genet 16:1604–1618CrossRefPubMedGoogle Scholar
  37. Sreedharan J, Blair IP, Tripathi VB et al (2008) TDP-43 mutations in familial and sporadic amyotrophic lateral sclerosis. Science 319:1668–1672CrossRefPubMedGoogle Scholar
  38. Vance C, Rogelj B, Hortobagyi T et al (2009) Mutations in FUS, an RNA processing protein, cause familial amyotrophic lateral sclerosis type 6. Science 323:1208–1211CrossRefPubMedGoogle Scholar
  39. Velculescu VE, Zhang L, Vogelstein B, Kinzler KW (1995) Serial analysis of gene expression. Science 270:484–487CrossRefPubMedGoogle Scholar
  40. Wendt S, Dedeoglu A, Speer O, Wallimann T, Beal MF, Andreassen OA (2002) Reduced creatine kinase activity in transgenic amyotrophic lateral sclerosis mice. Free Radic Biol Med 32:920–926CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Michel Guipponi
    • 1
    • 7
  • Qiao-Xin Li
    • 2
    • 3
  • Lavinia Hyde
    • 1
  • Tim Beissbarth
    • 4
  • Gordon K. Smyth
    • 4
  • Colin L. Masters
    • 3
  • Hamish S. Scott
    • 5
    • 6
  1. 1.Division of Molecular MedicineThe Walter and Eliza Hall Institute of Medical ResearchParkvilleAustralia
  2. 2.Department of PathologyUniversity of MelbourneParkvilleAustralia
  3. 3.The Mental Health Research InstituteParkville3052Australia
  4. 4.Division of BioinformaticsThe Walter and Eliza Hall Institute of Medical ResearchParkvilleAustralia
  5. 5.Department of Molecular PathologyThe Centre for Cancer Biology, Institute of Medical and Veterinary Science and The Hanson InstituteAdelaideAustralia
  6. 6.Adelaide Cancer Research Institute, The School of MedicineUniversity of AdelaideAdelaideAustralia
  7. 7.Division of Medical GeneticsUniversity Hospital of GenevaChêne-BourgSwitzerland

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