Induction of Cell Stress in Neurons from Transgenic Mice Expressing Yellow Fluorescent Protein: Implications for Neurodegeneration Research

Comley, Laura H.; Wishart, Thomas M.; Baxter, Becki; Murray, Lyndsay M.; Nimmo, Ailish; Thomson, Derek; Parson, Simon H.; Gillingwater, Thomas H.

doi:10.1371/journal.pone.0017639

Cited by 25 publications

(21 citation statements)

References 23 publications

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“…Similar observations were made with VN-βS and VN-γS (not shown). Fusion to VN approximately doubles the size of αS which may contribute to these alterations, as observed also with intact (full-length) fluorophores and corresponding fusion proteins [56][57][58]. These elevated levels of VN-αS protein were detected at early time points after transfection and sustained high throughout the duration of the experiments.…”

Section: Fusion Of Vn To Synuclein Results In Aberrant Protein Levelssupporting

confidence: 55%

Dimerization propensities of Synucleins are not predictive for Synuclein aggregation

Eckermann

Kügler

Bähr

2015

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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Aggregation and fibril formation of human alpha-Synuclein (αS) are neuropathological hallmarks of Parkinson's disease and other synucleinopathies. The molecular mechanisms of αS aggregation and fibrillogenesis are largely unknown. Several studies suggested a sequence of events from αS dimerization via oligomerization and pre-fibrillar aggregation to αS fibril formation. In contrast to αS, little evidence suggests that γS can form protein aggregates in the brain, and for βS its neurotoxic properties and aggregation propensities are controversially discussed. These apparent differences in aggregation behavior prompted us to investigate the first step in Synuclein aggregation, i.e. the formation of dimers or oligomers, by Bimolecular Fluorescence Complementation in cells. This assay showed some Synuclein-specific limitations, questioning its performance on a single cell level. Nevertheless, we unequivocally demonstrate that all Synucleins can interact with each other in a very similar way. Given the divergent aggregation properties of the three Synucleins this suggests that formation of dimers is not predictive for the aggregation of αS, βS or γS in the aged or diseased brain.

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Section: Fusion Of Vn To Synuclein Results In Aberrant Protein Levelssupporting

confidence: 55%

Dimerization propensities of Synucleins are not predictive for Synuclein aggregation

Eckermann

Kügler

Bähr

2015

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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“…However, a recent study suggested that data from these mice should be interpreted with caution (Comley et al, 2011). The authors showed that YFP expression causes upregulation of stress markers in the spinal cord, and increases neurofilament staining in distal motor axons at the neuromuscular junction in otherwise normal animals.…”

Section: Discussionmentioning

confidence: 99%

Absence of SOD1 leads to oxidative stress in peripheral nerve and causes a progressive distal motor axonopathy

Fischer

Asress

et al. 2012

Experimental Neurology

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Oxidative stress is commonly implicated in the pathogenesis of motor neuron disease. However, the cause and effect relationship between oxidative stress and motor neuron degeneration is poorly defined. We recently identified denervation at the neuromuscular junction in mice lacking the antioxidant enzyme, Cu, Zn-superoxide dismutase (SOD1) (Fischer et al., 2011). These mice show a phenotype of progressive muscle atrophy and weakness in the setting of chronic oxidative stress. Here, we investigated further the extent of motor neuron pathology in this model, and the relationship between motor pathology and oxidative stress. We report preferential denervation of fast-twitch muscles beginning between 1 and 4 months of age, with relative sparing of slow-twitch muscle. Motor axon terminals in affected muscles show widespread sprouting and formation of large axonal swellings. We confirmed, as was previously reported, that spinal motor neurons and motor and sensory nerve roots in these mice are preserved, even out to 18 months of age. We also found preservation of distal sensory fibers in the epidermis, illustrating the specificity of pathology in this model for distal motor axons. Using HPLC measurement of the glutathione redox potential, we quantified oxidative stress in peripheral nerve and muscle at the onset of denervation. SOD1 knockout tibial nerve, but not gastrocnemius muscle, showed significant oxidation of the glutathione pool, suggesting that axonal degeneration is a consequence of impaired redox homeostasis in peripheral nerve. We conclude that the SOD1 knockout mouse is a model of oxidative stress-mediated motor axonopathy. Pathology in this model primarily affects motor axon terminals at the neuromuscular junction, demonstrating the vulnerability of this synapse to oxidative injury.

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“…Moreover, it was also reported that fluorescent proteins expressed in cells per tissues can modify their molecular composition and cellular activities. [22][23][24][25] These results suggest that GFP marker gene is not suitable for human gene therapy. Now, we are analyzing the effect of neonatal gene therapy for MLD using self-complementary AAV 9 vector expressing only ASA for clinical trial.…”

Section: Discussionmentioning

confidence: 99%

Long-term correction of biochemical and neurological abnormalities in MLD mice model by neonatal systemic injection of an AAV serotype 9 vector

Miyake

Asakawa

et al. 2014

Gene Ther

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As both the immune system and the blood-brain barrier (BBB) are likely to be developmentally immature in the perinatal period, neonatal gene transfer may be useful for the treatment of lysosomal storage disease (LSD) with neurological involvements such as metachromatic leukodystrophy (MLD). In this experiment, we examined the feasibility of single-strand adeno-associated viral serotype-9 (ssAAV9)-mediated systemic neonatal gene therapy of MLD mice. ssAAV9 vector expressing human arylsulfatase A (ASA) and green fluorescent protein (GFP) (ssAAV9/ASA) was injected into the jugular vein of newborn MLD mice. High levels of ASA expression were observed in the muscle and heart for at least 15 months. ASA was continuously secreted into plasma without development of antibodies against ASA. Global gene transfer into the brain and spinal cord (SC), across the BBB, and long-term ASA expression in the central nervous system were detected in treated mice. Significant inhibition of the accumulation of sulfatide (Sulf) in the brain and cervical SC was confirmed by Alcian blue staining and biochemical analysis of the Sulf content. In a behavior test, treated mice showed a greater ability to traverse narrow balance beams than untreated mice. These data clearly demonstrate that MLD mice model can be effectively treated through neonatal systemic injection of ssAAV9/ASA.

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Induction of Cell Stress in Neurons from Transgenic Mice Expressing Yellow Fluorescent Protein: Implications for Neurodegeneration Research

Cited by 25 publications

References 23 publications

Dimerization propensities of Synucleins are not predictive for Synuclein aggregation

Dimerization propensities of Synucleins are not predictive for Synuclein aggregation

Absence of SOD1 leads to oxidative stress in peripheral nerve and causes a progressive distal motor axonopathy

Long-term correction of biochemical and neurological abnormalities in MLD mice model by neonatal systemic injection of an AAV serotype 9 vector

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