SourceData5_JBG007_pMAPK

Published: 30 December 2016| Version 2 | DOI: 10.17632/v7y3nvmd9v.2
Contributors:
Miriam Matamales,
Zala Skrbis,
J Bertran-Gonzalez

Description

\Data type and description: mapping of p-MAPK immunoreactive neurons in different corticl and striatal regions after 20 minutes of action sequence performance. All young and aged animals were rapidly anesthetized and trasncardially fixed while performing the test (day 18). Coronal sections corresponding to Bregma ~1.7, 1.1 and 0.15 mm (A-P) were collected in each animal, and were counterstained phospho-Thr202/Tyr-204-p44/42-MAPK antibody. Samples were brought to the spinning disk confocal microscope, and high resolution mosaics of the full cortical and striatal regions were obtained for each identified (left [L] or right [R]) hemisection. Images were analyzed through quantitative fluorescence in ImageJ2/Fiji. All p-MAPK immunoreactive neurons were identified in each image using thresholding and segmentation. Resulting ROIs were superimposed against the original, unmodified signal, and measures of neuronal count (Neuron #), position (X, Y coordinates in pixels), and Area (in px2) and p-MAPK intensity were obtained for each identified neuron (pixel size: 0.625 um/px). A total of 63114 activated neurons were quantified. \NOTE: Section in Subject R4 had damaged DLS

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Steps to reproduce

\Experiment summary: 8 Aged (A1-8; 20-22 months old) and 8 Young (Y1-8; 2-3 months old) mice (C57-Bl6) were submitted to instrumental training (increasing random ratio schedule of reinfocement). CRF: constant reinforcement; RR5: random ratio 5; RR10: random ratio 10; RR20: random ratio 20. Active lever counterbalanced (Left [L] or Right [R]). \Penalising regime: from day 10 of training, a penalising protocol was introduced (pen5). From day 13, this regime was increased to pen7. Pen5: If the last 5 actions are not LP, probability of reward delivery is reseted; Pen7: If the last 7 actions are not LP, probability of reward delivery is reseted. \Test: on Day 18, exercise was the same as Day 17, pellets were not limited. After 20 minutes, behaving mice were rapidly anaesthetised and intracardiacally perfused (see methods)