Fig. 3
From: CREB controls cortical circuit plasticity and functional recovery after stroke
hM4Di/CREB inhibition of cortical neurons after stroke blocks motor recovery. a Schematic shows the process by which DREADD hM4Di-CREB when activated by CNO (30 min before behavioral tasks) can lead to selective inactivation of neurons in motor cortex in which CREB is induced. b Immunohistochemical staining in peri-infarct M1 in mice transfected with hM4Di-CREB/eGFP (green, infected cells) overlapping with NeuN staining (purple) 4 weeks after stroke. Scale bar = 50 μm. Stereological quantification of motor cortex M1 CREB-induced cells (CREB-eGFP+ cells) relative to the whole motor cortex (NeuN+ cells). c Stereological quantification of two immediate early genes, pCREB (phospho-CREB) and Zif268, in hM4Di/CREB-induced neurons in M1 after CNO or Saline administration during pasta handling motor task. ** P < 0.05,***P < 0.005, one way ANOVA followed by Bonferroni post hoc test n = 4. Eight brain sections for each mouse. d, f Gridwalking tasks of forelimb function in gait. Y axis is percentage of the number of footfaults of the right (affected) forelimb contralateral to the stroke. e, g Pasta handling tasks. Y axis is the percentage of left forelimb adjustments (unaffected forepaw) relative to right forepaw (stroke-affected forepaw). d, e CREB gain of function. In the absence of CNO, Stroke + hM4Di-CREB induction in peri-infarct M1 produces a significant recovery in forelimb function over 3–4 weeks after stroke, respectively in pasta handling adjustments and gridwalking, as with studies in Fig. 2b, d. n = 13 for stroke CREB CNO, n = 10 for CREB, and n = 9 for stroke CREB. f, g Selective inactivation with hM4Di-CREB of neurons in motor cortex (CNO administration) blocks motor recovery compared to mice with stroke + control virus either in grid walking or in pasta handling adjustments (**P < 0.05, ***P < 0.005, two way ANOVA followed by Bonferroni post hoc test). h, i Pasta handling graphs show motor recovery induced by hM4Di-CREB induction in the same mice in the absence of CNO (washout, mice tested day after CNO administration) at 5 and 11 weeks after stroke. n = 12 for control virus CNO, n = 11 for CREB CNO, and n = 10 for stroke control virus CNO (h). i Individual animals in pasta handling performance after CNO administration and after washout and 5 and 11 weeks after stroke. Gray bar shows the average performance of stroke-only mice, taken from data in Fig. 1c–e
