Iron accumulation is widely thought to be detrimental for cellular survival. It is well known that the iron-catalyzed Fenton reaction can covert mitochondrial-related H
2O
2 to the highly reactive hydroxyl radical (·OH) that can damage intracellular molecules such as DNA, proteins and lipids. Accordingly, brain regions that have a high burden of iron accumulation are likely to be more susceptible to oxidative stress. The situation is further aggravated in DA neurons as iron can promote the oxidation of dopamine and facilitate the formation of dopamine quinone as well as the neurotoxic 6-OHDA (Hare and Double
2016). Iron can also enhance the aggregation of α-synuclein, which is particularly toxic to DA neurons (Levin et al.
2011). Given these, one could therefore readily appreciate the impact of iron accumulation on DA neuronal survival. However, it remains unclear whether iron accumulation represents a cause or consequence of neurodegeneration in PD (Daugherty and Raz
2015). Notably, several studies have revealed the accumulation of Fe
3+ (rather than the more reactive Fe
2+) in the SN of PD brains compared to controls (Dusek et al.
2015). Specifically, Mössbauer spectroscopy can distinguish between Fe
2+, which generates free radicals via the Fenton reaction, and Fe
3+ which is usually found stored in ferritin. Using this technique, Galazka-Friedman conducted a study in PD individuals and age-matched controls but found no evidence of Fe
2+ in the SN, suggesting that iron is stored as the more inert Fe
3+ form in ferritin (Galazka-Friedman et al.
2012). Similarly, our results showed increased Fe
3+ in the SN of DMT1 mice compared to WT controls, as detected using Perls staining. Finally, a study that involved MPTP-treated monkeys showed a reduction in TH-positive cells within a week of injection, but increase in iron levels was only observed after 4.5 months (He et al.
2003). This again suggests that iron accumulation may not be a direct cause of neuronal death, and could explain why accumulation of iron is generally observed but not always present in cases of neurodegeneration (Dashtipour et al.
2015). This may also explain why DMT1-expressing mice do not develop any overt signs of disease-associated phenotype despite exhibiting robust accumulation of iron, especially in the SN when fed with iron-supplemented diet. The lack of neurotoxicity in the transgenic mice could also be due in part to the upregulation of Parkin, which was observed in DMT1 mice treated with iron-supplemented diet. Parkin is a multifunctional ubiquitin ligase that is widely regarded to be a broad spectrum neuroprotectant capable of protecting the cells against a plethora of toxic insults (Zhang et al.
2015). Interestingly, a recent study by Roth and colleagues demonstrated that Parkin regulates metal transport via promoting proteasomal degradation of DMT1B (Roth et al.
2010). Consistent with this, Parkin overexpression affords considerable protection to cells treated with manganese (Higashi et al.
2004) and iron (this study). In light of these findings, it was intuitive for us to investigate whether the ablation of Parkin expression would promote a PD phenotype in DMT1-expressing mice. As described above, the double-mutant mice also fail to exhibit robust signs of Parkinsonism, even when treated with iron-supplemented diet until they reached 18 months of age. This reinforces the suggestion that iron accumulation may not be the culprit in PD. Supporting this, we have recently shown that low concentrations of iron can mitigate manganese induced cytotoxicity rather than having deleterious effects (Tai et al.
2016). Unlike iron, chronic manganese exposure is well documented to be the cause of motoric disturbances known as manganism (Chen et al.
2015). Curiously, even when treated with manganese-enriched diet, DMT1/Parkin KO double-mutant mice appear largely normal, although they tend to display lower motor performance on a rotarod in the initial months following treatment. Thus, despite lending itself as a “gene (i.e., Parkin KO)—environmental (i.e., DMT1-mediated accumulation of metal ions)” model of PD, the double-mutant mice are relatively resistant to neurodegeneration. This is somewhat reminiscent of the scenario with the triple Parkin/PINK1/DJ-1 knockout mice, which exhibit no evidence of Parkinsonism (Kitada et al.
2009). Notwithstanding this, DMT1/Parkin KO double-mutant mice are more vulnerable to 6-OHDA-induced neurotoxicity. This appears to be brought about by the combined effects of DMT1 overexpression and Parkin gene ablation as mice expressing DMT1 alone do not exhibit significantly different motoric phenotype from WT mice when treated with 6-OHDA. Similarly, Parkin-deficient mice alone are not more sensitive to the toxin (Perez et al.
2005).