Journal: The Journal of Biological Chemistry
Article Title: O-GlcNAc transferase couples nutrient availability to synaptic plasticity in paraventricular neurons to regulate satiety
doi: 10.1016/j.jbc.2025.111124
Figure Lengend Snippet: Nutritional state differentially modulates synaptic transmission in WT and OGT-KO neurons. A , representative trace showing suppression of spontaneous excitatory synaptic events following NBQX application in WT PVN αCaMKII neurons. The red arrow represents a typical excitatory spike. B – D , representative Western blots and quantification of total and surface GluA1 expression in primary cortical neurons. WT values were normalized to 1. OGT-KO total GluA1 = 1.282 ± 0.068; surface GluA1 = 1.629 ± 0.064. E–G , representative Western blots and quantification of total and surface GluA2 expression. WT values were normalized to 1. OGT-KO total GluA2 = 0.7101 ± 0.081; surface GluA2 = 0.5138 ± 0.075. H–J , representative Western blots and quantification of total and surface GluA3 expression. WT values were normalized to 1. OGT-KO total GluA3 = 0.6691 ± 0.032; surface GluA3 = 0.3935 ± 0.044. Western blot quantitative analyses were performed using the Wilcoxon signed-rank test. All Western blot data expressed as mean ± SEM. K , representative sEPSC traces in WT neurons under hungry ( top ) and fed ( bottom ) conditions. L , sEPSC frequency in WT neurons comparing hungry vs fed states ( p = 0.021). M , sEPSC amplitude in WT neurons comparing hungry vs fed states (NS, p = 0.851). N , sEPSC decay tau in WT neurons comparing hungry vs fed states (NS, p = 0.142). O , Representative sEPSC traces in OGT-KO neurons under hungry ( top ) and fed ( bottom ) conditions. P , sEPSC frequency in OGT-KO neurons comparing hungry vs fed states (NS, p = 0.095). Q , sEPSC amplitude in OGT-KO neurons comparing hungry vs fed states (NS, p = 0.309). R , sEPSC decay tau in OGT-KO neurons comparing hungry vs fed states (NS, p = 0.547). All sEPSC data ( panels K–R ) were obtained from WT (n = 5) and OGT-KO (n = 5) mice under hungry and fed conditions (n = 5 each condition). Statistical comparisons using Mann-Whitney non-parametric test. Data presented as mean ± SEM. Blue bars: WT neurons ( dark blue = hungry, light blue = fed); yellow bars: OGT-KO neurons ( light yellow = hungry, bright yellow = fed). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, NS, non-significant.
Article Snippet: Membranes were blocked in 5% non-fat milk prepared in TBS-T and incubated overnight at 4 °C with primary antibodies against OGT (Proteintech, 11576-2-AP; 1:2000), HSP70 (Proteintech, 10995-1-AP; 1:10,000), GluA1 (NeuroMab, N355/1; 5μg/blot), GluA2 (NeuroMab, L21/32; 5μg/blot), or GluA3 (Almone labs, AGC-010-GP; 1:1000).
Techniques: Transmission Assay, Western Blot, Expressing, MANN-WHITNEY
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