​(Fig.1A).1A). Our previous observations in 3-month-old B6eGFPChAT mice (Nagy and Aubert 2012) revealed enhanced VAChT protein expression and here, we confirm that at 6 months of age, VAChT overexpression is sustained. The expression of VAChT in B6eGFPChAT mice was compared with B6 controls using Western blot analysis to detect cholinergic immunoreactivity in various regions of the central nervous system. Western blot targeting VAChT revealed a diffuse doublet at the predicted size of 70 kDa (Fig. ​(Fig.1B).1B). Quantification Inhibitors,research,lifescience,medical of the VAChT band intensity revealed a significant two- to threefold increase

of VAChT protein in B6eGFPChAT compared with B6 control mice (Fig. ​(Fig.1C).1C). The enhanced level of VAChT protein was found in the cortex (t(4) = 8.752; P = 0.001), striatum (t(4) = 4.494; P = 0.046), and hippocampal formation (t(4) = 5.323; P = 0.006) (Fig. ​(Fig.1D).1D). Western blots and quantification of ChAT (Fig. ​(Fig.1D1D and E) and Inhibitors,research,lifescience,medical CHT (Fig. ​(Fig.1F1F and G) revealed no significant Inhibitors,research,lifescience,medical change in protein expression

in any of the regions that were analyzed. Figure 1 VAChT protein is overexpressed in B6eGFPChAT mice. (A) RP23-268L19-EGFP transgene is expressed throughout the central nervous system of B6eGFPChAT mice including cortical, striatal, and hippocampal regions (green). Punctate VAChT immunoreactivity is present … B6eGFPChAT mice exhibit unaltered motor function Inhibitors,research,lifescience,medical and coordination To measure the effect of increased VAChT on peripheral motor function, we first assessed forelimb grip strength using a digital tension gauge. B6eGFPChAT mice produced a peak tension of 0.268 kg which was not found to be significantly different from B6 control mice that produced 0.260 kg of peak tension (t(18) = 0.416; P = 0.682) (Fig. ​(Fig.2A).2A).

In addition, no learn more statistical difference was found between B6eGFPChAT and B6 control mice when measuring wire hang fatigue (two-way repeated measures ANOVA revealed no significant Inhibitors,research,lifescience,medical genotype factor, F(1,36) = 0.052; P = 0.822, and the expected trial factor, F(2,36) = 11.04; P < 0.001) (Fig. ​(Fig.2B)2B) or total hanging time performance (t(18) = 0.229; P = 0.822) (Fig. ​(Fig.22C). Figure crotamiton 2 Neuromuscular function and coordination in B6eGFPChAT mice. (A) Maximum forelimb grip strength measured in B6eGFPChAT (N = 11) and B6 control mice (N = 9). (B and C) Time spent hanging upside-down from a wire grid in each of three consecutive trials (B) … We considered that the effect of VAChT overexpression might only be detectable during activities combining endurance, fine motor coordination, and balance. As such, performance on the rotarod was assessed through the latency to fall off the rotating cylinder. Both B6eGFPChAT and B6 control mice improved significantly from trial 1 to trial 10 (two-way repeated measures ANOVA trial factor, F(9,162) = 8.653; P < 0.