Synchrotron radiation-based FTIR spectro-microscopy of the brainstem of the hSOD1 G93A rat model of amyotrophic lateral sclerosis
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Pathological mechanisms in amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disease, are still poorly understood. One subset of familial ALS cases is caused by mutations in the metallo-enzyme copper–zinc superoxide dismutase (SOD1), increasing the susceptibility of the SOD1 protein to form insoluble intracellular aggregates. Here, we employed synchrotron radiation-based Fourier transform infrared spectroscopy and microscopy to investigate brainstem cross-sections from the transgenic hSOD1 G93A rat model of ALS that overexpresses human-mutated SOD1. We compared the biomacromolecular organic composition in brainstem tissue cross-sections of ALS rats and their non-transgenic littermates (NTg). We demonstrate that the proteins and especially their antiparallel β-sheet structure significantly differed in all three regions: the facial nucleus (FN), the gigantocellular reticular nucleus (GRN) and the trigeminal motor nucleus (TMN) in the brainstem tissue of ALS rats. The protein levels varied between different brainstem areas, with the highest concentration observed in the region of the FN in the brainstem tissue of NTg animals. Furthermore, the concentration of lipids and esters was significantly decreased in the TMN and FN of ALS animals. A similar pattern was detected for choline and phosphate assigned to nucleic acids with the highest concentrations in the FN of NTg animals. The spectroscopic analysis showed significant differences in phosphates, amide and lipid structure in the FN of NTg animals in comparison with the same area of ALS rats. These results show that the hG93A SOD1 mutation causes metabolic cellular changes and point to a link between bioorganic composition and hallmarks of protein aggregation.
KeywordsAmyotrophic lateral sclerosis SOD1 Brainstem FTIR Biomolecular composition Protein aggregation
We thank the ALBA synchrotron light source for the beamtime allocation. TD’s work was carried out with financial support of the ALBA in-house research. This work was also supported by the Ministry of education, science and technological development of the Republic of Serbia (MESTD RS), Grant no. III41005 and the Biostruct-X project no. 5977. We thank Dr. Martin Kreuzer for help during the FTIR analysis. We declare all competing interests in relation to their work. The authors declare that they have no competing interests.
Conceptualization: TD, SS and PRA. Investigation: TD and SS. Formal analysis: TD. Visualization: TD and SS. Writing—original draft: TD. Writing—review and editing: TD.
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