Figure 7.

TS:YSOG3-PKMζ degrades rapidly in neuronal cell bodies and dendritic spines after glycine cLTP. A, Transfected neurons stimulated with glycine were fixed and immunolabeled for PSD95. TS:YSOG3-PKMζ YFP fluorescence (green pseudocolor in magnified images) and PSD95 immunofluorescence (red pseudocolor) were imaged. Stimulated neurons show increased YFP fluorescence and TS:YSOG3-PKMζ colocalization with PSD95 (yellow pseudocolor). B, Schematic indicating how transfected neurons were treated with BILN for 24 h and then imaged once per hour for 10 h after BILN washout. C, Stimulated neuron YFP fluorescence decreases quickly over time, indicating that PKMζ is rapidly degraded. D, Unstimulated neuron YFP fluorescence decreases slowly in the 10 h following BILN washout, indicating that basal PKMζ is slowly degraded. n = 52 stimulated dendritic spines; n = 51 unstimulated dendritic spines. PKMζ produced at the basal level remains more stable over time. E, YFP quantification in neuronal cell bodies (n = 5 stimulated cell bodies; n = 5 unstimulated cell bodies) and (F) dendritic spines (n = 55 stimulated dendritic spines; n = 52 unstimulated dendritic spines). *p < 0.05 (unpaired two-tailed t test). **p < 0.01 (unpaired two-tailed t test). ***p < 0.001 (unpaired two-tailed t test). Error bars indicate SEM. NS, Not significant). Scale bars: A, whole cell, 20 μm; magnified pseudocolor, 2 μm; C, D, 20 μm.