Mosharov EV, Larsen KE, Kanter E, Phillips KA, Wilson K, Schmitz Y, Krantz DE, Kobayashi K, Edwards RH, Sulzer D. Interplay between cytosolic dopamine, calcium, and alpha-synuclein causes selective death of substantia nigra neurons. Neuron. 2009 Apr 30;62(2):218-29. PubMed.
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Ann Romney Center for Neurologic Diseases, BWH and HMS
This paper by Mosharov et al. takes a very contemporary look at an old question: Why are the dopaminergic neurons of the substantia nigra pars compacta (SNc) more vulnerable in Parkinson disease than those in the neighboring ventral tegmental area (VTA)? By considering the role of α-synuclein and the unique calcium-dependent pacemaking activity of SNc neurons recently shown by Jim Surmeier’s group, David Sulzer and colleagues propose a novel and provocative model for the molecular basis of the unique susceptibility of the SNc in disease.
By demonstrating not only an enhanced vulnerability of nigral neurons to L-DOPA-induced toxicity, but also providing a pharmacologic basis for their observations, they put forth an interesting hypothesis regarding the selective accumulation of neuromelanin in this brain region, as well.
One caveat that must be openly considered, and is likewise raised in the thoughtful commentary by Jim Surmeier that accompanies the Mosharov report, is that the endogenous levels of cytoplasmic dopamine cannot be measured by currently available technologies. Thus, the authors are only able to follow the kinetics of L-DOPA and dopamine metabolism in cultures treated with micromolar concentrations of L-DOPA, thus raising the cytoplasmic dopamine levels into detectable range. As the authors are first to point out, the levels of L-DOPA provided to their neuronal cultures are about three orders of magnitude above those that occur following therapeutic administration of L-DOPA to Parkinson’s patients and likely seven orders of magnitude above those seen in normal human brain. One wonders whether the differences in aromatic amino acid decarboxylase (AADC) activities suggested by these data at 100 μM L-DOPA would hold true in the presence of only 5 nM L-DOPA. It seems possible that at potentially saturating concentrations of L-DOPA one observes differences that would not exist at the extremely lower physiological or even therapeutic levels of substrate. This potential confound is also consistent with the classical view that tyrosine hydroxylase and not AADC is the rate-limiting step in dopamine synthesis, in vivo. With the argument that differential AADC activities could play a determining role in cytoplasmic dopamine levels, one then places AADC at the bottleneck. Even with this caveat in mind, however, it must be acknowledged that the methods applied here define the state-of-the-art for measuring dopamine and the technical hurdles already surpassed by this group are not trivial.
One last note regarding the authors’ hypothesis that calcium may regulate the activity of AADC; in Figure 6 of the report the authors nicely show the differential response of SNc and VTA neurons to inhibitors of the L-type calcium channels. While not specifically emphasized by the authors, I found it quite interesting that the calcium chelator BAPTA-AM, or use of cadmium, had significant inhibitory effects on L-DOPA metabolism in both culture systems. I would interpret these data as very consistent with their model of a potent, calcium-dependent effect on dopamine synthesis and/or metabolism. Further work will be required to specifically evaluate the influence of calcium, whether it be calcium that specifically passes across L-type calcium channels, or not. These data, along with the incrimination of α-synuclein in the mechanism L-DOPA- or dopamine-dependent toxicity in this culture system, are likely to influence the future experiments of many of us who study the etiology of Parkinson disease.
View all comments by Matthew LaVoieNational Institute on Aging
One of the difficulties with the longstanding idea that dopamine is a contributor to neuronal cell loss in Parkinson disease (PD) and related disorders is that it appears neither to be necessary nor sufficient to cause cell death. Dopamine can’t be necessary for cell death as there are non-dopaminergic neurons that are lost throughout the disease process. And dopamine on its own is unlikely to be sufficient to cause neurodegeneration, as there appear to be dopamine neurons that are relatively spared in PD.
This paper by Mosharov et al. highlights the idea that simple rules are unlikely to explain complex diseases. The authors address the sufficiency argument by showing that dopamine contributes to toxicity only in some contexts. Calcium is also highlighted and is a pretty good candidate for a modifier, especially given the relatively poor calcium buffering capacity of dopaminergic nigral neurons that is probably intrinsically related to the physiological role they have to serve. Again, it seems unlikely that calcium alone is either necessary or sufficient to explain the complex patterns of relative vulnerability in PD, but one could imagine that both neurotransmitter identity added to a specific physiological role of this group of neurons might impact their long-term survival.
The most interesting candidate identified here, albeit briefly in one figure, is α-synuclein. We know from other evidence that α-synuclein can be a causal factor in PD and that the difference between no disease and an aggressive, progressive diffuse Lewy body disorder is determined by α-synuclein expression over a twofold range. The data here that α-synuclein influences nigral neuron survival without shifting intracellular dopamine levels strongly implicate this protein as having a detrimental effect on cells downstream of other toxic stressors. This is consistent with a number of previous observations, including the relative resistance of α-synuclein null mice to different stressors and the proposal that there might be a toxic pairing of α-synuclein and dopamine.
But why α-synuclein would have this property of cascading damage is not at all obvious. Why even would neurons make a protein at relatively high levels if that protein is a perennial bad apple? It is interesting that smaller, simpler organisms such as fruit flies do apparently fine without any synuclein genes. Perhaps α-synuclein has some important benefit to the function of a neuron in the context of either longer-lived species (we know that aging is another contributory factor in PD) or specific physiological needs (which might relate in some way to the calcium-mediated effects seen here, although that link seems immediately tenuous). I think this question might be approached either by understanding what the normal of function of α-synuclein is and by identifying why exactly it is toxic to cells under some conditions. The Mosharov paper is an important start to this process.
View all comments by Mark CooksonEmory University
This is a great paper. The idea of cytosolic dopamine being toxic has been unsubstantiated dogma in the field for years. There have been numerous indirect indications of free dopamine contributing to the vulnerability of the substantia nigra neurons, but we had to take leaps of faith when discussing this. The lingering comment was always "if we only had a way of measuring cytoplasmic dopamine in nigral neurons." Well, the Sulzer laboratory has utilized an elegant combination of techniques to achieve this.
More importantly, they used the technique to help unify several of the hot topics in the field, namely, calcium regulation and α-synuclein expression. Application of this technique in other transgenic models related to Parkinson disease should help advance the field even further.
From a patient standpoint, these findings do suggest that there could be new treatments on the horizon. The better we can manage dopamine inside and outside the dopamine neuron, the better we can manage the disease progression and therapeutic treatment.
View all comments by Gary Miller