Extended Data Fig. 4: The effects of inhibitor against ALDH1a1 or shRNA against ALDH2 on LTP-induced GABA increase.
From: ALDH2-mediated GABA biosynthesis regulates hippocampal LTP and learning adaptability
a, The ALDH-mediated GABA biosynthetic pathway and inhibitors. b, Representative confocal microscopy images of immunostaining for ALDH1a1 (cyan) and DAPI (blue) fluorescence in the CA1 of Vglut2-Cre;Ai9 mice. Images are representative of three independent experiments. Scale bar, 20 μm. c, Normalized iGABA intensities in CA1 PNs with baseline stimulation or TBS (Vehicle: n = 29 Ctrl, 29 LTP; CM37: n = 26 Ctrl, 29 LTP). The PNs were treated with vehicle or CM37 (10 µM). Data normalized to Vehicle-Ctrl group. d, The LTP induced by TBS with 10 µM CM37 absent or present in the internal solution (Vehicle, n = 5 neurons; CM37, n = 6 neurons). Top, representative electrophysiological traces at the indicated time. e, Schematic illustration depicting viral shALDH2 constructs (ALDH2* indicates an shRNA-resistant ALDH2) and experimental configuration. Panel e is created with BioRender.com. f, Quantitative analysis of ALDH2 mRNA by RT-qPCR from CA1 after injection of AAV virus (n = 4 mice for each group). g, Confocal images depicting the infection of virus (EGFP) and ALDH2 (red) in CA1. Images are representative of three independent experiments. Scale bars, 50 μm. h, Normalized iGABA intensities of in CA1 PNs with baseline stimulation or TBS. The PNs were from Vglut2-Cre mice receiving intra-CA1 injection of AAV-DIO-scramble (shScr), AAV-DIO-shALDH2 (shALDH2) or AAV-DIO-shALDH2-ALDH2* (shALDH2 Rescue) (n values from left to right = 37, 34, 30 and 30). Data were normalized to values from the shScr-Ctrl group. i, Normalized 15N-GABA intensities in CA1 PNs with baseline stimulation or TBS. The PNs were incubated with 15N-ABAL (n values from left to right = 32, 35, 33 and 26) (n values from left to right = 32, 35, 33 and 26). Data normalized to shScr-Ctrl group. Unless otherwise stated, data were collected from 3 mice. Data are represented as mean ± SEM. Statistics were calculated by two-way ANOVA (c), one-way ANOVA with Tukey’s multiple comparison post hoc test (h) or unpaired two-tailed t-test (f, i). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ns, not significant.
