Figure 2.
Brain homogenates from GBA1+/L444P mice showed reduced GCase activity and selective reduction of vGLUT1 compared with GBA1+/+mice. A, GCase activity assay to evaluate enzymatic activity of GCase in 2-month-old GBA1+/L444P mice in the cortex (independent t test: t(10) = 7.406, ****p < 0.0001), striatum (t(10) = 3.023, *p = 0.0128), hippocampus (t(10) = 6.870, ****p < 0.0001), and the midbrain (t(10) = 4.593; ***p < 0.0010) expressed as nm 4-MU/mg protein. N = 6 for all groups. B, Representative immunoblot analyses of hippocampal brain lysates for GCase, endogenous total α-syn, p-α-syn, Homer1, TUJ1, SNAP25, VAMP2, and syntaxin-1 expression. C, Quantitation of immunoblots; vinculin was used as the loading control to normalize expression. Independent t test for GCase (t(4) = 1.093, p = 0.3360, N = 3 for both groups, one GBA1+/+ outlier removed) and Mann–Whitney U test for endogenous total α-syn (W = 4, p = 0.9999, N = 3 for both groups). An independent t test was used for p-α-syn (t(9) = 2.808; *p = 0.0205, N = 6 GBA1+/+ and N = 5 GBA1+/L444P). D, Analysis of vGLUT1 (independent t test: t(5) = 6.686; **p = 0.0011, N = 4 GBA1+/+ and N = 3 GBA1+/L444P, one GBA1+/+ outlier removed based on Grubbs' test), Homer1 (t(4) = 1.481; p = 0.2127, N = 3 both groups), and TUJ1 (t(9) = 0.05656, p = 0.9561, N = 6 GBA1+/+ and N = 5 GBA1+/L444P) expression. E, Analysis of SNAP25 (independent t test: t(5) = 0005117, p = 0.9996, N = 4 GBA1+/+ and N = 3 GBA1+/L444P), VAMP2 (t(4) = 1.433, p = 0.2251, N = 3 in both groups, one GBA1+/+ outlier removed), and syntaxin-1a (t(6) = 0.3713, p = 0.7232, N = 5 GBA1+/+ and N = 3 GBA1+/L444P) expression. All immunoblots were run with 3-6 individual mice. For all graphs, error bars indicate SEM. The total synuclein data failed the test for normality and graphed as a scatter plot without a bar using a Mann–Whitney test. *p < 0.05. **p < 0.01. ***p < 0.001. ****p < 0.0001.