Journal: iScience

Article Title: Conditional deletion of TRPC1 channel modulates synaptic plasticity, long term depression, and memory extinction in Fragile X syndrome mice

doi: 10.1016/j.isci.2025.113085

Figure Lengend Snippet: Representative immunostaining of surface and internalized GluA1 subunit of AMPAR in hippocampal neurons (A) Representative immunostaining shows the distribution of surface and internalized GluA1-AMPAR subunit in hippocampal neurons in FXS C1 vs. FXS C1-cKO stimulated or not with 100 μM of DHPG. (B) Quantification of the internalization index (internalized/total) in both groups, showing a significant increase of GluA1 internalization in FXS C1 compared to FXS C1-cKO, 60 min post DHPG stimulation. 10–20 images were collected in each n, ( n = 4, each n represents one coverslip of hippocampal neuron culture from one mouse), two-way ANOVA, F (1,13) = 5.100, p = 0.0418, genotype effect). (C) Representative and quantitative Western blot of samples collected from cultured hippocampal neurons, showing a stable total GluA1 protein expression in both groups (FXS C1 n = 5, FXS C1-cKO n = 5, each n represents one coverslip of hippocampal neuron culture from one mouse, Student’s t test). All experiments were done in the presence of 50 μM picrotoxin and 10 μM D-AP5. Values are means ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ns not significant.

Article Snippet: 2.5 μg (1/65 dilution) of an antibody directed at the N-terminus of the GluA1 receptor subunit, anti-GluA1 (Cell Signaling #13185) diluted in cell neuronal culture medium (Neurobasal medium supplemented with glutamine, B27, 10% FBS) was applied on live hippocampal neurons for 15 min, before being washed out with neurobasal media.

Techniques: Immunostaining, Western Blot, Cell Culture, Expressing