Data

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Distribution of Fast blue-positive neurons.

<p>(A) The sagittal brain atlas indicating the level of the section in B (purple line; i-viii). (B) Representative pictures of the brain sections in the cerebral cortex (i, ii and iii), thalamus and hypothalamus (iv), midbrain (v), pons (vi) and medulla oblongata (vii and viii). The neurons projecting to the contralesional spinal cord at C7-8 were labeled with Fast blue. Infarct is indicated with a red line and the observed area of Fast blue-positive cells are indicated with a purple line. The names of the brain areas are abbreviated as follows: Rostral forelimb area (RFA), caudal forelimb area (CFA), secondary sensory area (S2), hypothalamic area (Hyt), mesencephalic tegmental area (Mes. Teg.), red nucleus (RN), nucleus reticularis pontis (NRPN) mesencephalic trigeminal nuclei (Mes. Trig.), deep cerebellar nuclei (Cbll), nucleus reticularis gigantocellularis (NRGi), nucleus reticularis parvocellularis (NRPa), spinal trigeminal nuclei (Sp. Trig.), vestibular nuclei (Vest), dorsal medullary reticular nucleus (MdD), and ventral medullary reticular nucleus (MdV). Scale bar = 1 mm.</p

Summary of the histological changes induced by rehabilitative skilled forelimb training.

<p>Summary of the histological changes induced by rehabilitative skilled forelimb training.</p

Experimental design.

<p>(A) After completion of skill acquisition training, rats received photothrombotic stroke (PT) in the CFA. Rat with rehabilitative training (PT + Rehab) carried out a skilled forelimb reaching task 100 times per day, 5 days per week for 4 weeks. Fast blue was injected 3 to 4 days after final behavioral testing. (B) Fast blue was injected into the lower cervical spinal cord (C7-8) in the contralesional side. Orange areas indicate forelimb areas (divided into RFA and CFA). Black circles indicate infarct induced by photothrombosis.</p

The effect of skilled forelimb training on functional recovery following stroke.

<p>(A) Results of the cylinder test. Rats showed a significant decrease in use of the preferred forelimb following stroke. Preferred forelimb use recovered in both groups. (B) Results of the ladder walk test. Stroke increased foot fault in the contralesional forelimb and hindlimb. Although the amount of foot fault decreased slightly in the contralesional forelimb 4 weeks following stroke, sustained disabilities were observed both in the contralesional forelimb and hindlimb. (C) Results of the skilled forelimb reaching test. Stroke caused severe disability in the skilled forelimb reaching task. While rats without rehabilitative training showed almost no improvement in motor performance, rats with rehabilitative training achieved nearly full recovery. (D) Results of the staircase test. Stroke caused impaired motor performance in the contralesional side. The impairment persisted until 4 weeks following stroke in both the untrained and trained rats. Unexpectedly, the trained rats showed a significant decline in motor performance in the ipsilesional (less affected) side. Repeated measures two-way ANOVA, followed by post-hoc test: Tukey’s multiple comparison test for the comparisons between the different time points. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 vs. pre-stroke in the same group, Sidak’s multiple comparison test for the comparison between groups at each time point. ##p<0.01, ####p<0.0001 PT vs. PT + Rehab.</p

Tracer injection validation.

<p>(A) Representative photo of the Fast blue injection in the spinal cord. Scale bar = 1 mm. (B) Injection area of Fast blue at the C7 and C8 spinal levels was not significantly different between groups. (C) There were no significant differences between groups in the location of the injection centroid at the C7 and C8 spinal levels.</p

Distribution of Fast blue-positive neurons in the cerebral cortex.

<p>Representative pictures of Fast blue labeled brain sections in the rostral forelimb area (A) and secondary sensory area (B) of the PT rats and the PT + Rehab rats. The ipsilesional hemisphere is located on the left side. Scale bars = 1 mm.</p

The effect of skilled forelimb training on the spinal projections from the cerebral cortex.

<p>The numbers of Fast blue-positive cells were analyzed in the ipsilesional (Ipsi) and contralesional (Contra) rostral forelimb area (A: RFA), and in the caudal forelimb area (B: CFA) and secondary sensory area (C: S2). Rehabilitative skilled forelimb training increased the number of Fast blue-positive cells in the ipsilesional RFA and the S2. Unpaired t-test: *p<0.05, **p<0.01 PT vs. PT + Rehab.</p

The effect of skilled forelimb training on the spinal projections from the brainstem.

<p>The numbers of Fast blue-positive cells were analyzed in the (A) Hypothalamic area (Hyt), (B) mesencephalic tegmental area (Mes. Teg.), (C) red nucleus (RN), (D) nucleus reticularis pontis (NRPN) (E) mesencephalic trigeminal nuclei (Mes. Trig.), (F) deep cerebellar nuclei (Cbll), (G) nucleus reticularis gigantocellularis (NRGi), (H) nucleus reticularis parvocellularis (NRPN), (I) spinal trigeminal nuclei (Sp. Trig.), (J) vestibular nuclei (Vest), (K) dorsal medullary reticular nucleus (MdD), and (L) ventral medullary reticular nucleus (MdV). Rehabilitative skilled forelimb training did not induce significant changes in any areas of the hypothalamus, midbrain, pons, cerebellum, or medulla oblongata.</p