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. 2016 Dec 23;5:e17197. doi: 10.7554/eLife.17197

Figure 4. Translational control of auditory imprinting by eIF2α.

(a) After 2 hr imprinting training, IMM and MNM were punched out for western blot analysis. The ratio of phosphorylated eIF2α (p-eIF2α) and non-phosphorylated eIF2α was measured in controls and after training in MNM (left) and IMM (right) brain tissue. Trained chicks (n = 7) exhibited decreased eIF2α phosphorylation compared to the untrained (n = 6) in MNM but not in IMM. Representative western blots are shown below each panel. * indicates p<0.05 from unpaired Mann-Whitney test. (b) Left, drugs injected for targeting the eIF2α pathway. Right, schematic effect of pharmacological manipulations on the eIF2α pathway. (c) Auditory (left) but not visual (right) imprinting is blocked by Sal003 injection (n = 12) compared to controls injected with vehicle (n = 9). (d) Auditory imprinting (left) was enhanced in chickens injected with the PKR inhibitor PKRi (n = 26), compared to controls injected with saline vehicle (n = 14). On the other hand, PKRi (n = 26) had no effect on visual imprinting (right), compared to saline injection (n = 14). (e) Auditory imprinting (left) but not visual imprinting (right) was enhanced by ISRIB administration (n = 11) compared to controls injected with vehicle (n = 13). Bar plots represent mean and SEM, * indicates p<0.05 from unpaired t-test.

DOI: http://dx.doi.org/10.7554/eLife.17197.009

Figure 4—source data 1. Western blots of p-eIF2α/ total eIF2α ratio and behavioral pharmacology after targeting the eIF2α pathway.
DOI: http://dx.doi.org/10.7554/eLife.17197.010
elife-17197-fig4-data1.xlsx (13.7KB, xlsx)
DOI: 10.7554/eLife.17197.010

Figure 4.

Figure 4—figure supplement 1. Sal003 increases eIF2α phosphorylation.

Figure 4—figure supplement 1.

Western blots for p-eIF2α and total eIF2α of brain samples obtained from chickens 2 hr after injecting Sal003 (S) or vehicle (V). Sal003 treatment increased eIF2α phosphorylation. * indicates p<0.05 from Mann-Whitney U Test.
DOI: http://dx.doi.org/10.7554/eLife.17197.011
Figure 4—figure supplement 2. ISRIB and Sal003 injection modulate translation in vivo.

Figure 4—figure supplement 2.

Sal003 and ISRIB can bi-directionally regulate protein synthesis. Both in MNM and IMM Sal003 reduces puromycin (green) incorporation while ISRIB enhances it. S6 (red) was used to localize cell somas. Bar plots show mean and SEM; different letters inside bars indicate statistically significant differences (p<0.05) between groups from Kruskal-Wallis test, Dunn’s multiple comparisons test.
DOI: http://dx.doi.org/10.7554/eLife.17197.012