.. Considering that SOD1 manages the accumulation of reactive species, and oxidative stress has been related to the neurodegenerative process in ALS (Basso et al. 2009), we analyzed the

time course of accumulation of nitrosative reactive species. We analyzed nitrotyrosine levels in gray and white matter of the spinal cord, as nitrosative reactive species have been early detected in the SOD1G93A mice. Besides, high levels of nitrotyrosine activate microglia to initiate synaptic stripping (Moreno-Lopez et al. 2011). The staining with antinitrotyrosine was practically absent in WT spinal cord; however, we observed that nitrotyrosine levels Inhibitors,research,lifescience,medical progressively increase in the gray matter and within the MN soma in SOD1G93A mice, reaching

significantly higher values at 2 months of age (Fig. 7). These data suggested that cholinergic alterations may occur earlier than peripheral neuromuscular detachment and consequently Inhibitors,research,lifescience,medical induced ER stress, but in parallel to the initial accumulation of oxidative reactive species. Tdp-43 Inhibitors,research,lifescience,medical Finally, considering that Tdp-43, also linked to ALS etiopathogenesis, is involved in multiple steps of RNA metabolism, including transcription, splicing, or transport of mRNA (Lagier-Tourenne and Cleveland 2010), as well as microRNA metabolism, and it has been recently Inhibitors,research,lifescience,medical shown to target ChAT mRNA as well, (Buratti et al. 2010) we wanted

to analyze its expression at early presymptomatic stages in this mouse model. Tdp-43 was found normally Selleckchem Hesperadin present in both nucleus and cytoplasm of the MNs in WT mice. In contrast, Tdp-43 was markedly overexpressed and accumulated Inhibitors,research,lifescience,medical in the nucleus but barely detected in the cytoplasm of spinal MNs in the SOD1G93A mice already at 1 month of age (Fig. 8). The same pattern was observed at 2 months. In contrast, from the symptomatic stage, by 3 months of age, Tdp-43 levels increased also in the cytoplasm of MNs and in the nucleus of surrounding glial cells within the spinal cord parenchyma. Figure 8 Tdp-43 is markedly accumulated in the MN nucleus of the transgenic SOD1G93A mice. Representative confocal microphotographs of single sections where a MN nucleus is present (DAPI staining in blue) showing ChAT (green) and Tdp-43 (red) immunolabeling, and … In conclusion, both the levels and localization many of Tdp-43 in all the spinal MNs are severely affected early in the presymptomatic stage in SOD1G93A mice, and parallels the development of cholinergic dysfunctions. Discussion Synaptic cholinergic dysfunction is a common feature of different neurodegenerative diseases, including ALS, but little is known regarding the possible relationship between ChAT abnormalities and the pathogenesis of MN degeneration.