Skip to main content
SpringerLink
Log in

Inverse correlation between the formation of mitochondria-derived vacuoles and Lewy-body-like hyaline inclusions in G93A superoxide-dismutase-transgenic mice

  • Original Paper
  • Published:
Acta Neuropathologica Aims and scope Submit manuscript

Abstract

In G93A mice, the most popular model of amyotrophic lateral sclerosis (ALS), neuronal Lewy-body-like hyaline inclusions (LBHIs) and mitochondria-derived vacuoles are observed in addition to motor neuron loss. Although LBHIs are thought to be toxic, the significance of the mitochondria-derived vacuoles has not been fully investigated. In this study, the relationship between the formation of these vacuoles and LBHIs was clarified statistically in the lumbar segment from two phyletic lines of G93A mice (G1L, G1H), using immunohistochemical methods. Furthermore, the distributions of vacuoles and LBHIs were examined in the pons including the facial nucleus, where pathological changes occur in ALS patients and G93A mice. Numerous vacuoles 2–3 μm in diameter were detected in the neuropil of the lumbar segment from G1L mice euthanatized approximately 3.5 months prior to the onset of the disease. Most of the vacuoles disappeared, but some became larger as the disease progressed. The number of vacuoles with a diameter exceeding 5 μm began to decrease after disease onset, while that of intra-neuritic LBHIs increased rapidly. There was a strong inverse correlation between the numbers of vacuoles and LBHIs in symptomatic mice (P<0.01; G1L, r=−0.91; G1H, r=−0.93). In the facial nucleus of G1L mice, where the number of motor neurons was significantly reduced, only a few LBHIs were detected along with prominent vacuole formation. In contrast, significantly more LBHIs with little vacuole formation were evident around the facial nucleus in G1L mice. Furthermore, the SOD1 immunoreactivity in vacuoles initially increased and then decreased after disease onset. Taken together, the present findings suggest that the mitochondria-derived vacuoles might prevent the formation of LBHIs by sequestering mutated SOD1 from the cytoplasm.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3.
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Barneoud P, Lolivier J, Sanger DJ, Scatton B, Moser P (1997) Quantitative motor assessment in FALS mice: a longitudinal study. Neuroreport 8:2861–2865

    Article  PubMed  CAS  Google Scholar 

  2. Bruening W, Roy J, Giasson B, Figlewicz DA, Mushynski WE, Durham HD (1999) Up-regulation of protein chaperones preserves viability of cells expressing toxic Cu/Zn-superoxide dismutase mutants associated with amyotrophic lateral sclerosis. J Neurochem 72:693–699

    Article  PubMed  CAS  Google Scholar 

  3. Brujin LI, Becher MW, Lee MK, Anderson KL, Jenkins NA, Copeland NG, Sisodia SS, Rothstein JD, Borchelt DR, Price DL, Cleveland DW (1997) ALS-linked SOD1 mutant G85R mediates damage to astrocytes and promotes rapidly progressive disease with SOD1-containing inclusions. Neuron 18:327–338

    Article  Google Scholar 

  4. Brujin LI, Houseweart MK, Kato S, Anderson KL, Anderson SD, Ohama E, Reaume AG, Scott RW, Cleveland DW (1998) Aggregation and motor neuron toxicity of an ALS-linked SOD1 mutant independent from wild-type SOD1. Science 281:1851–1854

    Article  Google Scholar 

  5. Cleveland DW, Liu J (2000) Oxidation versus aggregation—how do SOD1 mutants cause ALS? Nat Med 6(12):1320–1321

    Article  PubMed  CAS  Google Scholar 

  6. Dal Canto MC, Gurney ME (1994) Development of central nervous system pathology in a murine transgenic model of human amyotrophic lateral sclerosis. Am J Pathol 145:1271–1279

    PubMed  CAS  Google Scholar 

  7. Dal Canto MC, Gurney ME (1997) A low expressor line of transgenic mice carrying a mutant human Cu, Zn superoxide dismutase (SOD1) gene develops pathological changes that most closely resemble those in human amyotrophic lateral sclerosis. Acta Neuropathol 93:537–550

    Article  PubMed  CAS  Google Scholar 

  8. Deng H-X, Hentati A, tainer JA, Iqbal Z, Cayabyab A, Hung WY, Getsoff ED, Hu P, Herzfeldt B, Roos RP, Warner C, Deng G, Soriano E, Smyth C, Parge HE, Ahmed A, Roses AD, Hallwell RA, Pericak-Vance MA, Siddique T (1993) Amyotrophic lateral sclerosis and structural defects in Cu/Zn superoxide dismutase. Science 261:1047–1051

    Article  PubMed  CAS  Google Scholar 

  9. Durham HD, Roy J, Dong L, Figlewicz DA (1997) Aggregation of mutant Cu/Zn superoxide dismutase proteins in a culture model of ALS. J Neuropathol Exp Neurol 56(5):523–530

    Article  PubMed  CAS  Google Scholar 

  10. Fischer LR, Culver DG, Tennant P, Davis AA, Wang M, Castellano-Sanchez A, Khan J, Polak MA, Glass JD (2004) Amyotrophic lateral sclerosis is a distal axonopathy: evidence in mice and man. Exp Neurol 185:232–240

    Article  PubMed  Google Scholar 

  11. Guegan C, Vila M, Rosoklija G, Hays AP, Przedborski S (2001) Recruitment of the mitochondrial-dependent apoptotic pathway in amyotrophic lateral sclerosis. J Neurosci 21:6569–6576

    PubMed  CAS  Google Scholar 

  12. Gurney ME, Pu H, Chiu AY, Dal Canto MC, Polchow CY, Alexander DD, Caliendo J, Hentati A, Kwon YW, Deng HX, Chen W, Zhai P, Sufit RL, Siddique T (1994) Motor neuron degeneration in mice that express a human Cu, Zn superoxide dismutase mutation. Science 264:1772–1775

    Article  PubMed  CAS  Google Scholar 

  13. Haenggeli C, Kato AC (2002) Differential vulnerability of cranial motoneurons in mouse models with motor neuron degeneration. Neurosci Lett 335:39–43

    Article  PubMed  CAS  Google Scholar 

  14. Higgins CMJ, Jung C, Xu Z (2003) ALS-associated mutant SOD1G93A causes mitochondrial vacuolation by expansion of the intermembrane space and by involvement of SOD1 aggregation and peroxisomes. BMC Neurosci 4:16

    Article  PubMed  Google Scholar 

  15. Hirano A, Kurland LT, Sayre GP (1967) Familial amyotrophic lateral sclerosis. A subgroup characterized by posterior and spinocerebellar tract involvement and hyaline inclusions in the anterior horn cells. Arch Neurol 16:232–243

    PubMed  CAS  Google Scholar 

  16. Howland DS, Jiu J, She Y, Goad B, Maragakis NJ, Kim B, Erickson J, Kulik J, DeVito L, Psaltis G, DeGennaro LJ, Cleveland DW, Rothstein JD (2002) Focal loss of the glutamate transporter EAAT2 in a transgenic rat model of SOD1 mutant-mediated amyotrophic lateral sclerosis (ALS). Proc Natl Acad Sci USA 99:1604–1609

    Article  PubMed  CAS  Google Scholar 

  17. Inoue K, Fujimura H, Ogawa Y, Satoh T, Shimada K, Sakoda S (2002) Familial amyotrophic lateral sclerosis with a point mutation (G37R) of the superoxide dismutase 1 gene: a clinicopathological study. Amyotroph Lateral Scler Other Motor Neuron Disord 3:244–247

    PubMed  Google Scholar 

  18. Jaarsma D, Haasdijk ED, Grashorn JAC, Hawkins R, Dujin WV, Verspaget HW, London J, Holstege JC (2000) Human Cu/Zn superoxide dismutase (SOD1) overexpression in mice causes mitochondrial vacuolization, axonal degeneration, and premature motoneuron death and accelerates motoneuron disease in mice expressing a familial amyotrophic lateral sclerosis mutant SOD1. Neurobiol Dis 7:623–643

    Article  PubMed  CAS  Google Scholar 

  19. Jaarsma D, Rognoni F, Duijn WV, Verspaget HW, Haasdijk ED, Holstege JC (2001) CuZn superoxide dismutase (SOD1) accumulates in vacuolated mitochondria in transgenic mice expressing amyotrophic lateral sclerosis-linked SOD1 mutations. Acta Neuropathol 102:293–305

    PubMed  CAS  Google Scholar 

  20. Johnston JA, Ward CL, Kopito RR (1998) Aggresomes: a cellular response to misfolded proteins. J Cell Biol 143:1883–1898

    Article  PubMed  CAS  Google Scholar 

  21. Johnston JA, Dalton MJ, Gurney ME, Kopito RR (2000) Formation of high molecular weight complexes of mutant Cu,Zn-superoxide dismutase in a mouse model for familial amyotrophic lateral sclerosis. Proc Natl Acad Sci USA 97:12571–12576

    Article  PubMed  CAS  Google Scholar 

  22. Kato S, Takikawa M, Nakashima K, Hirano A, Cleveland DW, Kusaka H, Shibata N, Kato M, Nakano I, Ohama E (2000) New consensus research on neuropathological aspects of familial amyotrophic lateral sclerosis with superoxide dismutase 1 (SOD1) gene mutations: inclusions containing SOD1 in neurons and astrocytes. Amyotroph Lateral Scler Other Motor Neuron Disord 1:163–184

    Article  PubMed  CAS  Google Scholar 

  23. Kato S, Horiuchi S, Liu J, Cleveland DW, Shibara N, Nakashima K, Nagai R, Hirano A, Takikawa M, Kato M, Nakano I, Ohama E (2000) Advanced glycation endproduct-modified superoxide dismutase-1 (SOD1)-positive inclusions are common to familial amyotrophic lateral sclerosis patients with SOD1 gene mutations and transgenic mice expressing human SOD1 with a G85R mutation. Acta Neuropathol 100:490–505

    Article  PubMed  CAS  Google Scholar 

  24. Kato S, Sumi-Akamaru H, Fujimura H, Sakoda S, Kato M, Hirano A, Takikawa M, Ohama E (2001) Copper chaperone for superoxide dismutase co-aggregates with superoxide dismutase 1 (SOD1) in neuronal Lewy body-like hyaline inclusions: an immunohistochemical study on familial amyotrophic lateral sclerosis with SOD1 gene mutation. Acta Neuropathol 102:233–238

    PubMed  CAS  Google Scholar 

  25. Kato S, Saeki Y, Aoki M, Nagai M, Ishizaki A, Itoyama Y, Kato M, Asayama K, Awaya A, Hirano A, Ohama E (2004) Histological evidence of redox system breakdown caused by superoxide dismutase 1 (SOD1) aggregation is common to SOD1-mutated motor neurons in humans and animal models. Acta Neuropathol 107:149–158

    Article  PubMed  CAS  Google Scholar 

  26. Klivenyi P, Ferrante RJ, Mathews RT, Bogdanov MB, Klein AM, Andreassen OA, Mueller G, Wermer M, Kaddurah-Daouk R, Flint Beal M (1999) Neuroprotective effects of creatine in a transgenic animal model of amyotrophic lateral sclerosis. Nat Med 5:347–351

    Article  PubMed  CAS  Google Scholar 

  27. Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD (1997) The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science 275:1132–1136

    Article  PubMed  CAS  Google Scholar 

  28. Kong J, Xu Z (1998) Massive mitochondrial degeneration in motor neurons triggers the onset of amyotrophic lateral sclerosis in mice expressing a mutant SOD1. J Neurosci 18(9):3241–3250

    PubMed  CAS  Google Scholar 

  29. Liu X, Kim CN, Yang J, Jemmerson R, Wang X (1996) Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c. Cell 86:147–157

    Article  PubMed  CAS  Google Scholar 

  30. McHanwell S, Biscoe TJ (1981) The sizes of motoneurons supplying hindlimb muscles in the mouse. Proc R Soc Lond 213:201–216

    Article  CAS  Google Scholar 

  31. Mohajeri MH, Figlewicz DA, Bohn MC (1998) Selective loss of α motoneurons innervating the medial gastrocnemius muscle in a mouse model of amyotrophic lateral sclerosis. Exp Neurol 150:329–336

    Article  PubMed  CAS  Google Scholar 

  32. Nagai M, Aoki M, Miyoshi I, Kato M, Pasinelli P, Kasai N, Brown RH Jr, Itoyama Y (2001) Rats expressing human cytosolic copper-zinc superoxide dismutase transgenes with amyotrophic lateral sclerosis: associated mutations develop motor neuron disease. J Neurosci 21(23):9246–9254

    PubMed  CAS  Google Scholar 

  33. Nagano S, Ogawa Y, Yanagihara T, Sakoda S (1999) Benefit of a combined treatment with trientine and ascorbate in familial amyotrophic lateral scledosis model mice. Neurosci lett 265:159–162

    Article  PubMed  CAS  Google Scholar 

  34. Nagano S, Satoh M, Sumi H, Fujimura H, Tohyama C, Yanagihara T, Sakoda S (2001) Reduction of metallothioneins promotes the disease expression of familial amyotrophic lateral sclerosis mice in a dose-dependent manner. Eur J Neurosci 13:1363–1370

    Article  PubMed  CAS  Google Scholar 

  35. Nimchinsky EA, Young WG, Yeung G, Shah RA, Gordon JW, Bloom FE, Morrison JH, Hof PR (2000) Differential vulnerability of oculomotor, facial, and hypoglossal nuclei in G86R superoxide dismutase transgenic mice. J Comp Neurol 416:112–125

    Article  PubMed  CAS  Google Scholar 

  36. Olanow CW, Perl DP, DeMartino GN, McNaught KSP (2004) Lewy-body formation is an aggresome-related process: a hypothesis. Lancet Neurol 3:496–503

    Article  PubMed  Google Scholar 

  37. Paxinos G, Franklin KBJ (2001) The mouse brain in stereotaxic coordinates second edition. Academic Press, Figs 79–85

  38. Reed JC (1997) Cytochrome c: can’t live with it-can’t live without it. Cell 91:559–562

    Article  PubMed  CAS  Google Scholar 

  39. Ripps ME, Huntley GW, Hoff PR, Morrison JH, Gordon JW (1995) Transgenic mice expressing an altered murine superoxide dismutase gene provide an animal model of amyotrophic lateral sclerosis. Proc Natl Acad Sci USA 92:689–693

    Article  PubMed  CAS  Google Scholar 

  40. Rosen DR, Siddique T, Patterson D, Figlewicz DA, Sapp P, Hentati A, Donaldson D, Goto J, O’Regan JP, Deng H-X, Rahmani Z, Krizus A, Mckenna-Yasck D, Cayabyab A, Gaston SM, Berger R, Tanzi RE, Halperin JJ, Herzfeldt B, Van den Bergh R, Hung W-Y, Bird T, Deng G, Mulder DW, Smyth C, Laing NG, Soriano E, Pricak-Vance MA, Haines J, Rouleau GA, Gusella JS, Horritz HR, Brown RH (1993) Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis. Nature 362:59–62

    Article  PubMed  CAS  Google Scholar 

  41. Sasaki S, Ohsawa Y, Yamane K, Sakuma H, Shibata H, Nakano R, Kikugawa K, Mizutani T, Tsuji S, Iwata M (1998) Familial amyotrophic lateral sclerosis with widespread vacuolation and hyaline inclusions. Neurology 51:871–873

    PubMed  CAS  Google Scholar 

  42. Sasaki S, Warita H, Abe K, Iwata M (2004) Slow component of axonal transport is impaired in the proximal axon of transgenic mice with G93A mutant SOD1 gene. Acta Neuropathol 107:452–460

    Article  PubMed  Google Scholar 

  43. Sasaki S, Warita H, Murakami T, Abe K, Iwata M (2004) Ultrastructural study of mitochondria in the spinal cord of transgenic mice with a G93A mutant SOD1 gene. Acta Neuropathol 107:461–474

    Article  PubMed  Google Scholar 

  44. Shibata N, Hirano A, Kobayashi M, Siddique T, Deng HX, Hung WY, Kato T, Asayama K (1996) Intense superoxide dismutase-1 immunoreactivity in intracytoplasmic hyaline inclusions of familial amyotrophic lateral sclerosis with posterior column involvement. J Neuropathol Exp Neurol 55:481–490

    Article  PubMed  CAS  Google Scholar 

  45. Shimizu S, Narita M, Tsujimoto Y (1999) Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 399:486–487

    Google Scholar 

  46. Stathopulos PB, Rumfeldt JAO, Scholzs GA, Irani RA, Frey HE, Hallewell RA, Lepock JR, Meiering EM (2003) Cu/Zn superoxide dismutase mutants associated with amyotrophic lateral sclerosis show enhanced formation of aggregates in vitro. Proc Natl Acad Sci USA 100(12):7021–2026

    Article  PubMed  CAS  Google Scholar 

  47. Steiber A, Gonatas JO, Gonatas NK (2000) Aggregation of ubiquitin and a mutant ALS-linked SOD1 protein correlate with disease progression and fragmentation of the Golgi apparatus. J Neurol Sci 173:53–62

    Article  Google Scholar 

  48. Stephens B, Navarrete R, Guiloff RJ (2001) Ubiquitin immunoreactivity in presumed spinal interneurons in motor neurone disease. Neuropathol Appl Neurobiol 27:352–361

    Article  PubMed  CAS  Google Scholar 

  49. Sugai F, Yamamoto Y, Miyaguchi K, Zhou Z, Sumi H, Hamasaki T, Goto M, Sakoda S (2004) Benefit of valproic in suppressing disease progression of ALS model mice. Eur J Neurosci 20:3179–3183

    Article  PubMed  Google Scholar 

  50. Tsukihara T, Aoyama H, Yamashita E, Tomizaki T, Yamaguchi H, Shinzawa-Itoh K, Nakashima R, Yaono R, Yoshikawa S (1996) The whole structure of the 13-subunit oxidized cytochrome c oxidase at 2.8A. Science 272:1136–1144

    Article  PubMed  CAS  Google Scholar 

  51. Turner BJ, Lopes EC, Cheema SS (2003) Neuromuscular accumulation of mutant superoxide dismutase 1 aggregates in a transgenic mouse of familial myotrophic lateral sclerosis. Neurosci Lett 350:132–136

    Article  PubMed  CAS  Google Scholar 

  52. Turner BJ, Atkin JD, Farg MA, Zang DW, Rembach A, Lopes EC, Patch JD, Hill AF, Cheema SS (2005) Impaired extracellular secretion of mutant superoxide dismutase 1 associates with neurotoxicity in familial amyotrophic lateral sclerosis. J Neurosci 25:108–117

    Article  PubMed  CAS  Google Scholar 

  53. Urushitani M, Kurisu J, Takahashi R (2002) Proteasomal inhibition by misfolded mutant superoxide dismutase 1 induces selective motor neuron death in familial amyotrophic lateral sclerosis. J Neurochem 83:1030–1042

    Article  PubMed  CAS  Google Scholar 

  54. Wang J, Xu G, Borchelt DR (2002) High molecular weight complexes of mutant superoxide dismutase 1: age-dependent and tissue specific accumulation. Neurobiol Dis 9:139–148

    Article  PubMed  CAS  Google Scholar 

  55. Watanabe M, Dykes-Hoberg M, Culotta VC, Price DL, Wong PC, Rothstein JD (2001) Histological evidence of protein aggregation in mutant SOD1 transgenic mice and in amyotrophic lateral sclerosis neural tissues. Neurobiol Dis 8:933–941

    Article  PubMed  CAS  Google Scholar 

  56. Wong PC, Pardo CA, Borchelt DR, Lee MK, Copeland NG, Jenkins NA, Sisodia SS, Cleveland DW, Price DL (1995) An adverse property of a familial ALS-linked SOD1 mutation causes motor neuron disease characterized by vacuolar degeneration of mitochondria. Neuron 14:1105–1116

    Article  PubMed  CAS  Google Scholar 

  57. Yamashita S, Mita S, Kato S, Okado H, Ohama E, Uchino M (2003) Bcl-2 expression using retrograde transporter of adenoviral vectors inhibits cytochrome c-release and caspase-1 activation in motor neurons of mutant superoxide dismutase 1 (G93A) transgenic mice. Neurosci Lett 350:17–20

    Article  PubMed  CAS  Google Scholar 

  58. Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng TI, Jones DP, Wang X (1997) Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science 275:1129–1132

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

This study was supported in part by a Health and Labour Sciences Research Grant, Research on Measures for Incurable Disease, Ministry on Health, Labour and Welfare of Japan.

Author information

Authors and Affiliations

  1. Department of Neurology D-4, Osaka University Graduate School of Medicine, 2-2, Yamadaoka, Suita, 565-0871, Osaka, Japan

    Hisae Sumi, Seiichi Nagano, Harutoshi Fujimura & Saburo Sakoda

  2. Department of Neuropathology, Institute of Neurological Sciences, Faculty of Medicine, Tottori University, Yonago, Japan

    Shinsuke Kato

Authors
  1. Hisae Sumi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Seiichi Nagano
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Harutoshi Fujimura
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shinsuke Kato
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Saburo Sakoda
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hisae Sumi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sumi, H., Nagano, S., Fujimura, H. et al. Inverse correlation between the formation of mitochondria-derived vacuoles and Lewy-body-like hyaline inclusions in G93A superoxide-dismutase-transgenic mice. Acta Neuropathol 112, 52–63 (2006). https://doi.org/10.1007/s00401-006-0056-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00401-006-0056-x

Keywords

Search

Navigation