Abstract
Mutations in superoxide dismutase-1 (SOD1) cause a form of the fatal paralytic disorder amyotrophic lateral sclerosis (ALS), presumably by a combination of cell-autonomous and non–cell-autonomous processes. Here, we show that expression of mutated human SOD1 in primary mouse spinal motor neurons does not provoke motor neuron degeneration. Conversely, rodent astrocytes expressing mutated SOD1 kill spinal primary and embryonic mouse stem cell–derived motor neurons. This is triggered by soluble toxic factor(s) through a Bax-dependent mechanism. However, mutant astrocytes do not cause the death of spinal GABAergic or dorsal root ganglion neurons or of embryonic stem cell–derived interneurons. In contrast to astrocytes, fibroblasts, microglia, cortical neurons and myocytes expressing mutated SOD1 do not cause overt neurotoxicity. These findings indicate that astrocytes may play a role in the specific degeneration of spinal motor neurons in ALS. Identification of the astrocyte-derived soluble factor(s) may have far-reaching implications for ALS from both a pathogenic and therapeutic standpoint.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Rosen, D.R. et al. Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis. Nature 362, 59–62 (1993).
Deng, H.-X. et al. Amyotrophic lateral sclerosis and structural defects in Cu,Zn superoxide dismutase. Science 261, 1047–1051 (1993).
Gurney, M.E. et al. Motor neuron degeneration in mice that express a human Cu, Zn superoxide dismutase mutation. Science 264, 1772–1775 (1994).
Wong, P.C. et al. An adverse property of a familial ALS-linked SOD1 mutation causes motor neuron disease characterized by vacuolar degeneration of mitochondria. Neuron 14, 1105–1116 (1995).
Bruijn, L.I. et al. ALS-linked SOD1 mutant G85R mediated damage to astrocytes and promotes rapidly progressive disease with SOD1-containing inclusions. Neuron 18, 327–338 (1997).
Clement, A.M. et al. Wild-type nonneuronal cells extend survival of SOD1 mutant motor neurons in ALS mice. Science 302, 113–117 (2003).
Beers, D.R. et al. Wild-type microglia extend survival in PU.1 knockout mice with familial amyotrophic lateral sclerosis. Proc. Natl. Acad. Sci. USA 103, 16021–16026 (2006).
Boillee, S. et al. Onset and progression in inherited ALS determined by motor neurons and microglia. Science 312, 1389–1392 (2006).
Kostic, V., Jackson-Lewis, V., De Bilbao, F., Dubois-Dauphin, M. & Przedborski, S. Bcl-2: prolonging life in a transgenic mouse model of familial amyotrophic lateral sclerosis. Science 277, 559–562 (1997).
Custer, S.K. et al. Bergmann glia expression of polyglutamine-expanded ataxin-7 produces neurodegeneration by impairing glutamate transport. Nat. Neurosci. 9, 1302–1311 (2006).
Das, S. & Potter, H. Expression of the Alzheimer amyloid-promoting factor antichymotrypsin is induced in human astrocytes by IL-1. Neuron 14, 447–456 (1995).
Raoul, C. et al. Motoneuron death triggered by a specific pathway downstream of Fas. Potentiation by ALS-linked SOD1 mutations. Neuron 35, 1067–1083 (2002).
Wichterle, H., Lieberam, I., Porter, J.A. & Jessell, T.M. Directed differentiation of embryonic stem cells into motor neurons. Cell 110, 385–397 (2002).
Miles, G.B. et al. Functional properties of motoneurons derived from mouse embryonic stem cells. J. Neurosci. 24, 7848–7858 (2004).
Raoul, C. et al. Lentiviral-mediated silencing of SOD1 through RNA interference retards disease onset and progression in a mouse model of ALS. Nat. Med. 11, 423–428 (2005).
Nagai, M. et al. Rats expressing human cytosolic copper-zinc superoxide dismutase transgenes with amyotrophic lateral sclerosis: associated mutations develop motor neuron disease. J. Neurosci. 21, 9246–9254 (2001).
Lee, K.J., Mendelsohn, M. & Jessell, T.M. Neuronal patterning by BMPs: a requirement for GDF7 in the generation of a discrete class of commissural interneurons in the mouse spinal cord. Genes Dev. 12, 3394–3407 (1998).
Suurmeijer, A.J., van der Wijk, J., van Veldhuisen, D.J., Yang, F. & Cole, G.M. Fractin immunostaining for the detection of apoptotic cells and apoptotic bodies in formalin-fixed and paraffin-embedded tissue. Lab. Invest. 79, 619–620 (1999).
Sawada, M., Hayes, P. & Matsuyama, S. Cytoprotective membrane-permeable peptides designed from the Bax-binding domain of Ku70. Nat. Cell Biol. 5, 352–357 (2003).
Di Giorgio, F.P., Carrasco, M., Siao, M., Maniatis, T. & Eggan, K. Non–cell autonomous effect of glia on motor neurons in an embryonic stem cell–based ALS model. Nat. Neurosci. advance online publication, 15 April 2007 (doi:10.1038/nn1885).
Gong, Y.H., Parsadanian, A.S., Andreeva, A., Snider, W.D. & Elliott, J.L. Restricted expression of G86R Cu/Zn superoxide dismutase in astrocytes results in astrocytosis but does not cause motoneuron degeneration. J. Neurosci. 20, 660–665 (2000).
Wang, J. et al. Coincident thresholds of mutant protein for paralytic disease and protein aggregation caused by restrictively expressed superoxide dismutase cDNA. Neurobiol. Dis. 20, 943–952 (2005).
Kuo, J.J., Siddique, T., Fu, R. & Heckman, C.J. Increased persistent Na+ current and its effect on excitability in motoneurones cultured from mutant SOD1 mice. J. Physiol. (Lond.) 563, 843–854 (2005).
Zona, C., Pieri, M. & Carunchio, I. Voltage-dependent sodium channels in spinal cord motor neurons display rapid recovery from fast inactivation in a mouse model of amyotrophic lateral sclerosis. J. Neurophysiol. 96, 3314–3322 (2006).
Ricart, K. et al. Interactions between β-neuregulin and neurotrophins in motor neuron apoptosis. J. Neurochem. 97, 222–233 (2006).
Gould, T.W. et al. Complete dissociation of motor neuron death from motor dysfunction by Bax deletion in a mouse model of ALS. J. Neurosci. 26, 8774–8786 (2006).
Shin, J.Y. et al. Expression of mutant huntingtin in glial cells contributes to neuronal excitotoxicity. J. Cell Biol. 171, 1001–1012 (2005).
Howland, D.S. et al. 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 (2002).
Scheffler, B. et al. Functional network integration of embryonic stem cell-derived astrocytes in hippocampal slice cultures. Development 130, 5533–5541 (2003).
Lukas, T.J., Luo, W.W., Mao, H., Cole, N. & Siddique, T. Informatics-assisted protein profiling in a transgenic mouse model of amyotrophic lateral sclerosis. Mol. Cell. Proteomics 5, 1233–1244 (2006).
Silva, G.A., Feeney, C., Mills, L.R. & Theriault, E. A novel and rapid method for culturing pure rat spinal cord astrocytes on untreated glass. J. Neurosci. Methods 80, 75–79 (1998).
Chalazonitis, A., Kessler, J.A., Twardzik, D.R. & Morrison, R.S. Transforming growth factor α, but not epidermal growth factor, promotes the survival of sensory neurons in vitro. J. Neurosci. 12, 583–594 (1992).
Chalazonitis, A., Crain, S.M. & Kessler, J.A. Preferential cholinergic projections by embryonic spinal cord neurons within cocultured mouse superior cervical ganglia. Brain Res. 458, 231–248 (1988).
Rideout, H.J., Dietrich, P., Wang, Q., Dauer, W.T. & Stefanis, L. α-Synuclein is required for the fibrillar nature of ubiquitinated inclusions induced by proteasomal inhibition in primary neurons. J. Biol. Chem. 279, 46915–46920 (2004).
Kaji, K. & Matsuo, M. Aging of chick embryo fibroblasts in vitro. III. Polyploid cell accumulation. Exp. Cell Res. 119, 231–236 (1979).
Przedborski, S. et al. Increased superoxide dismutase activity improves survival of cultured postnatal midbrain neurons. J. Neurochem. 67, 1383–1392 (1996).
Pfaff, S.L., Mendelsohn, M., Stewart, C.L., Edlund, T. & Jessell, T.M. Requirement for LIM homeobox gene Isl 1 in motor neuron generation reveals a motor neuron-dependent step in interneuron differentiation. Cell 84, 309–320 (1996).
Ericson, J. et al. Pax 6 controls progenitor cell identity and neuronal fate in response to graded Shh signaling. Cell 90, 169–180 (1997).
Acknowledgements
The authors wish to thank H. Mitsumoto for his support, C. Henderson, W. Dauer, E. Schon and J. Krakauer for their comments and advice, and J. Jeon for assistance in preparing this manuscript. We thank T. Maniatis and K. Eggan for communicating their results before publication and for insightful discussions about the study. This study is supported by Muscular Dystrophy Association/Wings-over-Wall Street, the ALS Association, Project-ALS, US National Institutes of Health NS42269, NS38370, NS11766, AG 21617, ES013177 and DK58056, US Department of Defense Grant DAMD 17-03-1, the Parkinson's Disease Foundation and the Bernard and Anne Spitzer Fund. M.N. is the recipient of the Gardner's fellowship from the Muscular Dystrophy Association. T.M.J. is an investigator of the Howard Hughes Medical Institute. D.B.R. is the recipient of a Philippe Foundation grant for exchange programs between France and the United States.
Author information
Authors and Affiliations
Contributions
M.N., D.B.R. and T.N. conducted all experiments and participated in designing them and writing the manuscript; H.W. and A.C. assisted in the experiments and in writing the manuscript; T.M.J. and S.P. conducted the data analyses and wrote the manuscript; and S.P. supervised the project. H.W. and T.M.J. also provided critical reagents.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Fig. 1
Human SOD1 transgene expression is stable in both primary neuronal and astrocyte cultures over time and is harmless to astrocytes. (PDF 526 kb)
Supplementary Fig. 2
Mutant SOD1 astrocytes kill to the same extent NTg and mutated SOD1–expressing motor neurons. (PDF 453 kb)
Supplementary Fig. 3
Mutated SOD1 expression by astrocytes, motor neurons or both alters to the same extent motor neuron morphometry. (PDF 383 kb)
Supplementary Fig. 4
GABAergic interneuron morphometry is not affected by mutated SOD1-expressing astrocytes. (PDF 308 kb)
Supplementary Fig. 5
Glutamate, Fas ligand and several major cytokines and chemokines are not involved in the toxicity of mutant SOD1 astrocytes to motor neurons. (PDF 645 kb)
Supplementary Fig. 6
Mutated SOD1–expressing astrocytes kill motor neurons through a programmed cell death that can be caspase–independent. (PDF 424 kb)
Rights and permissions
About this article
Cite this article
Nagai, M., Re, D., Nagata, T. et al. Astrocytes expressing ALS-linked mutated SOD1 release factors selectively toxic to motor neurons. Nat Neurosci 10, 615–622 (2007). https://doi.org/10.1038/nn1876
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nn1876
This article is cited by
-
Desloratadine alleviates ALS-like pathology in hSOD1G93A mice via targeting 5HTR2A on activated spinal astrocytes
Acta Pharmacologica Sinica (2024)
-
Neuropathogenesis-on-chips for neurodegenerative diseases
Nature Communications (2024)
-
FUS-ALS hiPSC-derived astrocytes impair human motor units through both gain-of-toxicity and loss-of-support mechanisms
Molecular Neurodegeneration (2023)
-
Stem cell-derived brainstem mouse astrocytes obtain a neurotoxic phenotype in vitro upon neuroinflammation
Journal of Inflammation (2023)
-
Peripheral administration of nanomicelle-encapsulated anti-Aβ oligomer fragment antibody reduces various toxic Aβ species in the brain
Journal of Nanobiotechnology (2023)