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Figure 7. Knockdown of eIF3A, RPS17, and FUS blocks visual experience-dependent behavioral plasticity.(A) Protocol to evaluate the effect of eIF3A, FUS, and RPS17 knockdown on visual avoidance behavior. Tecta were electroporated with control MO or eIF3A MO + RPS17 MO + FUS MO and 2 days later were assayed for baseline visual avoidance behavior at −1 hr, exposed to VE for 4 hr and assayed for visual avoidance behavior 5 hr and 24 hr after the onset of VE. (B) VE-induced behavioral plasticity in control animals was detected at 5 hr and maintained at 24 hr. (C) Triple knockdown blocked behavioral plasticity. AI values were normalized to baseline at −1 hr (B–C). (B–C) CMO: AI = 1 ± 0.07, 1.32 ± 0.09, or 1.48 ± 0.09; n = 21, 27, or 31 animals for tests at −1 hr, 5 hr, or 24 hr; eIF3A MO + RPS17 MO + FUS MO: AI = 1 ± 0.10, 1.19 ± 0.08, or 1.18 ± 0.08; n = 22, 33, or 33 animals for tests at −1 hr, 5 hr, or 24 hr. *=p < 0.05, **=p < 0.01, the Steel-Dwass test was used for nonparametric multiple comparisons with control. Error bars represent ±SEM. (D) Schematic of visual experience-induced dynamics in protein synthesis machinery. By combining BONCAT and DiDBiT, we identified CPPs including master regulators that control gene expression or protein translation, such as eIF3A, FUS, and RPS17, and effector proteins that maintain plasticity in response to VE, such as cytoskeletal proteins. Functional analysis of select CPPs suggests that de novo synthesis of master regulators is required for experience-dependent plasticity. Our data suggest that the experience-dependent changes in the nascent proteome result from a combination of direct changes in synthesis of master regulators and effector proteins, and secondary effects downstream of differential synthesis of master regulators.10.7554/eLife.33420.021Figure 7—source data 1. Quantification of normalized AI values data presented in Figure 7B–C. Image published in: Liu HH et al. (2018) © 2018, Liu et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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