Adult male, Long Evans black-hooded rats (mean weight of 300 grams) were
first trained and tested on the skilled forelimb reaching task (Figure 1A ) as previously described.7 (link) This task requires fine digit movement and intact
motor and sensory neural pathways.10 Animals that successfully retrieved 15 out of 20 pellets during
3 consecutive training sessions received permanent middle cerebral artery occlusion
to impair the preferred forelimb as described.7 (link) After middle cerebral artery occlusion, animals continued to
use their impaired forelimb to reach, despite showing deficits in performing the
task. To reduce possible effects of rehabilitation through daily testing, animals
were tested Monday through Friday for the first week after stroke and once per week
until time of treatment, ie, 9 weeks after stroke. Any animal that improved
spontaneously to retrieve >10 pellets in any testing session before the
9-week treatment point was removed from the study.
Nine weeks after stroke, animals received the purified monoclonal function
blocking mouse anti-Nogo-A antibody 11C711 (n=6), an inactive control Ab
(IgG1 against wheat auxin; n=4), or no treatment (n=5). Antibody was delivered for 2
weeks using the intracerebroventricular route through an osmotic minipump (2ML2;
Alzet) as described.8 (link)After a 9-week period of behavioral testing after treatment, the
neuroanatomical tracer, biotin dextran amine (BDA), was injected into the forelimb
motor cortex contralateral to the stroke lesion as described.4 (link) Two weeks after BDA injection,
animals were perfused, brains were removed, cryosectioned, and processed for
BDA-positive crossing corticorubral fibers as described,4 (link) and stroke lesion analysis was performed using the
method of Kawamata et al.12 Because behavioral testing showed no difference between stroke only and
stroke/control Ab animals (see Results), data from these 2 groups of animals were
pooled for BDA-positive fiber analysis.
Personnel performing behavioral testing or histological analyses were
blinded to the treatments. For behavioral analysis, a repeated-measures ANOVA was
used to compare the rate of improvement on the skilled forelimb reaching task and a
1-way ANOVA with Tukey post hoc analysis was used to compare the mean success
scores. Midline crossing corticorubral fibers were analyzed using a Student
t test. Stroke lesion size was analyzed using a 1-way
ANOVA.
first trained and tested on the skilled forelimb reaching task (
motor and sensory neural pathways.10 Animals that successfully retrieved 15 out of 20 pellets during
3 consecutive training sessions received permanent middle cerebral artery occlusion
to impair the preferred forelimb as described.7 (link) After middle cerebral artery occlusion, animals continued to
use their impaired forelimb to reach, despite showing deficits in performing the
task. To reduce possible effects of rehabilitation through daily testing, animals
were tested Monday through Friday for the first week after stroke and once per week
until time of treatment, ie, 9 weeks after stroke. Any animal that improved
spontaneously to retrieve >10 pellets in any testing session before the
9-week treatment point was removed from the study.
Nine weeks after stroke, animals received the purified monoclonal function
blocking mouse anti-Nogo-A antibody 11C711 (n=6), an inactive control Ab
(IgG1 against wheat auxin; n=4), or no treatment (n=5). Antibody was delivered for 2
weeks using the intracerebroventricular route through an osmotic minipump (2ML2;
Alzet) as described.8 (link)After a 9-week period of behavioral testing after treatment, the
neuroanatomical tracer, biotin dextran amine (BDA), was injected into the forelimb
motor cortex contralateral to the stroke lesion as described.4 (link) Two weeks after BDA injection,
animals were perfused, brains were removed, cryosectioned, and processed for
BDA-positive crossing corticorubral fibers as described,4 (link) and stroke lesion analysis was performed using the
method of Kawamata et al.12 Because behavioral testing showed no difference between stroke only and
stroke/control Ab animals (see Results), data from these 2 groups of animals were
pooled for BDA-positive fiber analysis.
Personnel performing behavioral testing or histological analyses were
blinded to the treatments. For behavioral analysis, a repeated-measures ANOVA was
used to compare the rate of improvement on the skilled forelimb reaching task and a
1-way ANOVA with Tukey post hoc analysis was used to compare the mean success
scores. Midline crossing corticorubral fibers were analyzed using a Student
t test. Stroke lesion size was analyzed using a 1-way
ANOVA.
