Scientists may have answered the neurological chicken-and-egg question in the case of amyotrophic lateral sclerosis (ALS): Which comes first, the protein aggregates or the disease? The large clusters of mutant superoxide dismutase 1 (SOD1) that are a hallmark of some forms of ALS do not cause disease, but appear later on, according to a paper published online this week in PNAS. Scientists in the laboratory of David Borchelt at the University of Florida in Gainesville tracked SOD1 aggregation in three mouse models of ALS to show that the pathology began before the mutant enzyme aggregated. The work does not rule out a role for aggregated SOD1 in later disease progression, Borchelt said. The scenario is reminiscent of that in Alzheimer disease (AD). Research suggests that it is soluble Aβ, not the aggregated forms, that is most toxic in AD (see ARF related news story and ARF news series, Part 1, Part 2, and Part 3).

Inherited mutations in SOD1—of which more than 100 varieties have been reported—are responsible for approximately 20 percent of familial ALS cases, or 2 percent of ALS cases overall. Rodents overexpressing mutant human SOD1 are a common lab model for the disease, and both the rodents and people have aggregated SOD1 in their motor neurons. Scientists suspected the aggregation was a late-stage process, but the current work confirms it, Borchelt said: “What we have done is basically tied the package up.”

Borchelt, first author Celeste Karch, and colleagues followed three mouse lines, expressing mutant SOD1-G93A, G37R, or H46R/H48Q. Karch harvested spinal cords from animals at time points ranging from 30 days to end-stage, when animals are paralyzed, and measured the amount of detergent-insoluble, aggregated SOD1. Pathological features such as Golgi apparatus degradation and astrogliosis preceded the appearance of SOD1 clusters, confirming that the animals were already getting sick when SOD1 started to aggregate. The authors did not investigate other forms of the protein, such as soluble SOD1 clusters, which could still play a role in disease onset. In AD, for example, soluble, oligomeric Aβ attacks synapses (see ARF related news story and Berman et al., 2008) and is toxic to dendritic spines (see ARF related news story and Koffie et al., 2009).

The researchers also used their experiments to examine the link between aberrant disulfide bond formation and aggregation. SOD1 contains four cysteines, and in its mature, dimerized form each monomer contains an intramolecular disulfide bridge. It has been suggested that deviant, intermolecular disulfide bonds between SOD molecules can seed oligomerization (see ARF related news story and Banci et al., 2009). However, human SOD1 mutations that remove one or the other cysteine involved in the intermolecular bonding have been associated with familial ALS, and in previous work, Karch and Borchelt found that cysteine-free mutants could still aggregate (Karch and Borchelt, 2008). In the current study, the authors found that even when they used the reducing agent β-mercaptoethanol, which destroys disulfide bridges, when extracting tissue from G93A mice, the SOD1 aggregates remained. Borchelt suggests that aberrant disulfide bonding is concomitant with aggregation, but not required. “We cannot convince ourselves that there is cause and effect,” he said.

Karch did find that the majority of insoluble SOD1 was reduced, implying that it never formed, or lost, the normal disulfide bond. This was the case both in end-stage mice and in cultured human embryonic kidney cells overexpressing A4V, G37R, and G93A mutant SOD1. The mutant protein may be less likely to form the correct disulfide bridge, the authors suggest, remaining in an immature, aggregation-prone state.

The work rules out a role for SOD1 aggregates in causing disease, but does not clarify what they actually do. The aggregates could merely be markers for diseased cells that cannot maintain protein homeostasis, or they could accelerate progression of the illness when they appear. “I’m guessing they make it worse,” said Joan Valentine of UCLA, who has collaborated with Borchelt and communicated the current paper to PNAS.

Understanding how ALS progresses could be more important that comprehending how it starts, Valentine said, because disease progression is a better clinical target. Armed with the details of progression, “we could stop it in its tracks,” she said.

Borchelt’s next step will be to discover what, if anything, the aggregates do to damage cells. “We won’t know what aggregation does in disease until we are able to stop it and see if it makes a difference,” he said. To that end, Borchelt is searching for compounds that will block aggregation.—Amber Dance


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News Citations

  1. The Many Misdeeds of Aβ—Seizures and Axonal Transport Interference
  2. San Diego: Oligomers Live Up to Bad Reputation, Part 1
  3. San Diego: Oligomers Live Up to Bad Reputation, Part 2
  4. San Diego: Oligomers Live Up to Bad Reputation, Part 3
  5. PIP2 at the Core of Aβ Oligomers’ Synaptic Attack?
  6. Spine Shrinkers: Aβ Oligomers Caught in the Act
  7. No Metal, No Stability: Structure of Apo SOD1

Paper Citations

  1. . Oligomeric amyloid-beta peptide disrupts phosphatidylinositol-4,5-bisphosphate metabolism. Nat Neurosci. 2008 May;11(5):547-54. PubMed.
  2. . Oligomeric amyloid beta associates with postsynaptic densities and correlates with excitatory synapse loss near senile plaques. Proc Natl Acad Sci U S A. 2009 Mar 10;106(10):4012-7. PubMed.
  3. . Structural and dynamic aspects related to oligomerization of apo SOD1 and its mutants. Proc Natl Acad Sci U S A. 2009 Apr 28;106(17):6980-5. PubMed.
  4. . A limited role for disulfide cross-linking in the aggregation of mutant SOD1 linked to familial amyotrophic lateral sclerosis. J Biol Chem. 2008 May 16;283(20):13528-37. PubMed.

Further Reading


  1. . Amyloid beta-induced neuronal hyperexcitability triggers progressive epilepsy. J Neurosci. 2009 Mar 18;29(11):3453-62. PubMed.
  2. . Progressive aggregation despite chaperone associations of a mutant SOD1-YFP in transgenic mice that develop ALS. Proc Natl Acad Sci U S A. 2009 Feb 3;106(5):1392-7. PubMed.
  3. . Disulphide-reduced superoxide dismutase-1 in CNS of transgenic amyotrophic lateral sclerosis models. Brain. 2006 Feb;129(Pt 2):451-64. PubMed.
  4. . Disruption of fast axonal transport is a pathogenic mechanism for intraneuronal amyloid beta. Proc Natl Acad Sci U S A. 2009 Apr 7;106(14):5907-12. PubMed.
  5. . Synaptic transmission block by presynaptic injection of oligomeric amyloid beta. Proc Natl Acad Sci U S A. 2009 Apr 7;106(14):5901-6. PubMed.
  6. . Disulfide cross-linked protein represents a significant fraction of ALS-associated Cu, Zn-superoxide dismutase aggregates in spinal cords of model mice. Proc Natl Acad Sci U S A. 2006 May 2;103(18):7148-53. PubMed.

Primary Papers

  1. . Role of mutant SOD1 disulfide oxidation and aggregation in the pathogenesis of familial ALS. Proc Natl Acad Sci U S A. 2009 May 12;106(19):7774-9. PubMed.