Extended Data Fig. 3: The identification of glutamate-GABA pathway in CA1 neurons.
From: ALDH2-mediated GABA biosynthesis regulates hippocampal LTP and learning adaptability
a, Representative confocal microscopy images of immunostaining for GAD65 (cyan) and DAPI (blue) fluorescence in the CA1 of Vglut2-Cre;Ai9 mice. The CA1 PNs were labeled with tdT (red). Images are representative of three independent experiments. Scale bar, 20 μm. b, c, Representative MS spectra for 15N-GABA and normalized intensities (c) of intracellular 15N-GABA with preincubation of 15N-glutamate in CA1 interneurons and PNs (Interneurons, n = 29; PNs, n = 22). Data were normalized to values from interneurons. d-f, Normalized intracellular concentrations of iGABA (d), iGlutamate (e), and iABAL (f) in single CA1 PNs under 15N-glutamate treatment (n = 29 Ctrl, 34 LTP) versus untreated controls (n = 28 Ctrl, 33 LTP). Values were normalized to their respective untreated control groups. g, h, Representative MS spectra (g) and normalized intensities (h) of iGABA in single CA1 interneurons treated with vehicle or 3-MPA (Vehicle, n = 36; 3-MPA, n = 31). Data were normalized to values from the vehicle group. Unless otherwise stated, each dot represents one neuron and all neurons measured from 3 mice. Data are represented as mean ± SEM. Statistics were calculated by unpaired two-tailed t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
