Article Figures & Data
Figures
- Figure 1.
Animal performance during a fast lever-pressing task. After one session of continuous reinforcement with self-paced delivery of up to 30 food pellets (CRF30), animals (n = 7) were required to perform on a FR schedule, whereby eight lever presses resulted in delivery of a food pellet within a time contingency, which ranged from one-thousand to 1 s (FR8-1000s to FR8-1s). A, Scheme representing the behavioral setup and structure of the fast lever-pressing task, as well as the task acquisition as represented by the average number of lever presses for each day of training (F(9,52) = 22.59, p = 0.0009). B, Distance of all seven consecutive IPIs from the final covert target (F(2.155,12.93) = 4.638, p = 0.0283). C, Percentage of sequences containing the minimum frequency target of the last session (end-target: 7 IPIs < 1s, ∼8.0 Hz; F(12,91) = 2.765, p = 0.0030); mean ± SEM represented in all graphs.
- Figure 2.
RNA polymerase II RPB1 phosphorylation in the striatum of mice trained in a fast lever-pressing task. A, Immunoblot analysis of the total RPB1 CTD repeat YS2PTS5PS7, with indication of its hyperphosphorylated (II0) and hypophosphorylated (IIA) forms, Ser5P+-enriched RPB1 CTD and Ser2P+-enriched RPB1 CTD. B, Actin quantification across both phospho-isoforms. C, RNA polymerase II poising index (calculated as the quotient between the Ser5-P and Ser2-P RPB1 CTD phospho-isoforms) in the striatum of mice trained in the fast lever pressing. For both control and trained groups, n = 7; data as mean ± SEM; ***p < 0.005.
- Figure 3.
Enrichment of RNA polymerase II RPB1 CTD phosphorylation forms at IEGs in the striatum of mice trained in a fast lever-pressing task. A, Graphical representation (not to scale) of the relative position of primers used in ChIP-qPCR experiments (primers represented in orange). B, ChIP-qPCR analysis of total RPB1 CTD binding at Gapdh and Tubb5 (positive control targets) and Arc and Fos (IEGs); controls n = 5; trained n = 6. C, ChIP-qPCR % of input data as a ratio between the promoter and gene bodies of all genomic targets for the total RPB1 CTD repeat. Data as mean ± SEM.
- Figure 4.
Dynamics of Ser5P+- and Ser2P+-RPB1 CTD enrichment and resulting poising indices at IEGs in the striatum of mice trained in a fast lever-pressing task. ChIP-qPCR % of input data for Ser5P+-enriched RPB1 CTD and Ser2P+-enriched RPB1 CTD (A) and Ser5P/Ser2P RNA polymerase II RPB1 CTD ratios (poising indices) (B) at Gapdh and Tubb5 (positive control targets) and Arc and Fos (IEGs) in the striatum of mice trained in the fast lever-pressing task (controls n = 3; trained n = 4). Data as mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.005; n.s., p > 0.05.
- Figure 5.
RPB1 CTD poising index at IEGs in the striatum and its correlation with learning of a fast lever-pressing task. ChIP-qPCR % of input data for pooled Ser5P/Ser2P RNA polymerase II RPB1 CTD ratios (poising indices) at the promoters and gene bodies of positive control targets (Gapdh and Tubb5) and IEGs (Arc and Fos) in the striatum of (A) control and performance control mice (performance control animals were killed after completion of FR8-50s, roughly corresponding to a halfway point in the training regime), as well as mice fully trained in the fast lever-pressing task (controls n = 4; performance controls n = 4; trained n = 3); (B) performance control and fully trained mice grouped as, respectively, far from target and close to target, according to the distance to target value (i.e., distance of all seven consecutive IPIs from the final covert target) each animal presented at completion of FR8-50s or FR8-1s schedules, with far from target animals displaying a distance to target value over 0.6 and close to target animals a distance to target value under 0.6; and (C) mice trained in the fast lever-pressing task grouped according to the number of lever presses performed in their final training session, with low pressing animals finishing with under 250 lever presses and high pressing animals with over 250 lever presses. Data as mean ± SEM; *p < 0.05; **p < 0.01; n.s., p > 0.05.
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