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. 2017 Dec 4;6:e24192. doi: 10.7554/eLife.24192

Figure 6. Deletion of Shisa7 slows down initiation and decreases maintenance of LTP.

(a) Normalized EPSP amplitude over the time course of the LTP experiments shows a clear genotype effect (WT, n = 5 slices, n = 5 mice; Shisa7 KO, n = 6 slices, n = 6 mice). The arrow indicates the theta burst stimulation. Inset: Example of LTP effect on WT (left) and the Shisa7 KO (right) showing the EPSP shape during baseline (black) and after LTP induction (gray, red; last 10 minutes). (b) Normalized EPSP amplitude binned during 5 minute intervals early after LTP induction (0–5’) and in the maintenance phase (20–25’, 25–30’) shows a significant genotype effect. Individual data are shown in Figure 6—figure supplement 2. (c) Zoom-in of the first 2 minute after theta burst application (arrow) for normalized EPSP amplitude showed an immediate genotype effect (unpaired t-tests, *p<0.05; #p<0.1). Dotted lines show base-line EPSP amplitude. (d) Example of GluA1 surface staining (green; MAP, blue) of hippocampal neurons (WT, Shisa7 KO) after glycine-induced chemical LTP vs. non-stimulated (scale bar 20 µm). Insets are shown in Figure 6—figure supplement 2. (e) Quantification of number of surface GluA1 spots normalized to dendritic length from two independent experiments (n = 26–28 wells per group, based on average of 15–40 pictures per well) showed a significant effect of LTP in WT but not in Shisa7 KO neurons. Individual data are shown in Figure 6—figure supplement 2.

Figure 6.

Figure 6—figure supplement 1. Deletion of Shisa7 slows down initiation and decreases maintenance of LTP.

Figure 6—figure supplement 1.

(a) Normalized EPSP slope over the time course of the LTP experiments shows a clear genotype effect, with a reduction in Shisa7 KO, similar as observed for amplitude (cf. Figure 5)(WT, n = 5 slices, n = 5 mice; Shisa7 KO, n = 6 slices, n = 6 mice). The arrow indicates the theta burst stimulation. Inset: schematic of LTP induction protocol, with vertical bar representing extracellular current injections (action potential (AP) generation) and synaptic stimulation (EPSPs with a 3 ms delay) for 1 out of 3 theta bursts given. (b) Quantification of the normalized EPSP slope binned during 5 minute intervals early after LTP induction (0–5’) and in the maintenance phase (20–25’, 25–30’), showed a significant genotype effect only in the latter time points. (c) A zoom-in of the first 2 minutes after theta burst application (arrow) showed a trend for a genotype effect for normalized EPSP slope at this short time interval (unpaired t-tests, *p<0.05; #p<0.1).
Figure 6—figure supplement 2. Deletion of Shisa7 slows down initiation and decreases maintenance of LTP by affecting AMPAR recruitment.

Figure 6—figure supplement 2.

(a) Normalized EPSP amplitude (WT, n = 5 slices, n = 5 mice; Shisa7 KO, n = 6 slices, n = 6 mice mind that the last data point contains n = 5) binned during 5 minute intervals early after LTP induction (0–5’) and in the maintenance phase (20–25’, 25–30’) shows a significant genotype effect with individual data points indicated. (b) Enlarged examples of GluA1 surface staining (green; MAP, blue) from hippocampal neurons (WT, Shisa7 KO) after glycine-induced chemical LTP vs. non-stimulated as represented in Figure 6d (scale bar: 10 µm). (c) Normalized surface GluA1 boutons per µm dendrite after chemical LTP shows a significant genotype effect with individual data points indicated for data resulting from two independent experiments (batch 1, DIV16; batch 2, DIV14).