Fig 1: Altered molecular composition of AMPAR complexes in mHb neurons after adult ablation of GFRa1.(A, B) Immunoblots of synaptic protein fractions from mHb (A) and IPN (B) of WT, Het, and KO mice sacrificed 1 month after tamoxifen treatment probed for GluA1, 2, 3 and 4 as indicated. PSD95 was probed as loading control. Quantifications (± SEM) of GluA1 to 4 levels were corrected for PSD95 levels and normalized to levels in WT samples. N = 9–10 samples per group; 1-way ANOVA analysis followed by Tukey multiple comparison test; * P < 0.05; ** P < 0.01. (C, E) Immunoblots of synaptic protein fractions from mHb (C) and IPN (E) of WT, Het, and KO mice probed for GluA1 P-Ser831 and P-Ser845 as indicated and reported for total GluA1 and Tubulin as loading controls. (D, F) Quantifications (± SEM) of GluA1 P-Ser831 and P-Ser845 levels in mHb (D, N = 13 samples per group) and IPN (F, N = 14 samples per group) corrected for total GluA1 levels and normalized to levels in WT samples. Student t test; * P < 0.05. (G, I) PLA signals (red) for GluA1-GluA2 (G) and GluA1-GluA4 (I) complexes in coronal sections of mHb and core IPN from WT and KO mice. Counterstaining with DAPI appears in blue. Scale bars, 20 µm. (H, J) Quantification (± SEM) of PLA puncta for GluA1-GluA2 (H) and GluA1-GluA4 (J) complexes in mHb, core IPN and lateral IPN (IPL) from WT and KO mice (N = 5 mice per group). A total of 16 images from the mHb (8 dorsal and 8 ventral) and 18 images from the IPN (core IPN: 6 dorsal and 6 ventral; 6 lateral) were analyzed per mouse. Student t test; * P < 0.05. The data underlying this figure can be found at https://figshare.com/projects/Raw_Data_Fernandez-Suarez_et_al_2021/123406. GFRa1, glial cell–derived neurotrophic factor receptor alpha 1; HET, heterozygous; IPN, interpeduncular nucleus; KO, knockout; mHb, medial habenula; PLA, proximity ligation assay; PSD, postsynaptic density; WT, wild-type.
Supplier Page from Abcam for Anti-GluA4 antibody