Stroke is the leading cause of long-lasting disability in the United States and disproportionately affects adults in later life. Age-related decreases in dexterity and neural plasticity may contribute to the poorer prognosis of older stroke survivors, even following rehabilitative physical therapy. The goal of these dissertation studies is to determine how the cortical plasticity underlying motor skill learning, both before and after brain injury, changes in the aged brain. The general hypothesis of these studies is that age-related changes in motor performance and the limited ability to regain function following brain injury are associated with dysfunctional plasticity of the forelimb representation in the motor cortex. This hypothesis was tested in intact C57BL/6 mice by training them on a skilled reaching task and deriving intracortical microstimulation evoked motor cortical representations of the forelimb to determine training-induced changes in the function of the motor cortex. After ischemic lesions, age-dependencies in the effects of rehabilitative training in skilled reaching on forelimb motor cortical representations were investigated. Prior to injury, intact young and aged mice learned a skilled reaching task in similar time frames and with similar success rates. Training-induced reorganization in the young mouse motor cortex occurred in the caudal forelimb area, which is homologous to the primary motor cortex of primates. However, the rostral forelimb area, a potential premotor cortex, was larger in aged mice compared to young mice. Following focal ischemic lesions of the forelimb area of the sensorimotor cortex, aged mice had larger lesions and were more impaired than young mice, but both groups regained reaching ability after 9 weeks of rehabilitative training. Post-operative training resulted in plasticity of the rostral forelimb area in young mice, but we failed to see reorganization in the forelimb map of aged mice following rehabilitative training. These dissertation studies suggest that more severe brain damage in response to ischemia leads to poorer outcome in aged animals. Although the reorganization of motor cortex following initial skill learning and relearning following brain damage changes with age, the ability to learn motor tasks and improve function with rehabilitative training is maintained in healthy aging. / text

Seventy percent of stroke survivors experience loss of upper limb function after stroke and rehabilitative therapy is the only option to reduce impairments. However, uncertainty remains as to the optimal dose of therapy that should be prescribed. It has been suggested to report multiple parameters of dose, to increase standardization within the field, and to gain a better understanding of the dose-response relationship. This study investigated the automatic quantification of multiple dose parameters in a rat model of ischemic stroke, with rehabilitation paradigms whereby rats repeatedly grasp for food pellets to train their forelimb function. Starting 7 days post-stroke, groups of rats received 4, 8, or 12 rehabilitative training sessions for 10 days, practicing either high-quality (precision practice) or less skilled (mass practice) reaching movements. Pellet consumption was measured after each session and various metrics were analyzed using deep learning-based software (DeepLabCut, DLC) to represent parameters of dose intensity (number of reaches, paw path length) and session density (time on task). Functional outcome was assessed with the Montoya staircase task. Computer algorithms were validated against human analysis, demonstrating reach detection accuracy and reliability >80%. Interestingly, the number of training sessions did not alter the accumulated movement practice across rehabilitation, in either task. However, the number of sessions inversely affected training intensity, resulting in more forelimb use per session in rats with 4 sessions compared to 12 sessions. We found strong positive correlations between the number of reaches, time on task, paw path length, and pellets consumed in the precision practice, but only between reaches and pellets consumed in mass practice. This work demonstrates the quantification of multiple dose parameters using deep learning software and shows subtle differences between the two commonly used forelimb training tasks. Moreover, our data suggest that rehabilitative training at a frequency that is too high may negatively impact performance per session.

There is a paucity of data on neurochemical abnormalities and associated effects on cognition and motor performance in rats ontogenetically treated with quinpirole, a rodent model of dopaminergic hyperfunction. The objective of the current study was to analyze the cognitive and motor effects produced by ontogenetic administration of quinpirole, a dopamine D2/D3 receptor agonist. Past research from this laboratory has shown that ontogenetic quinpirole treatment sensitizes D2 receptors and produces a variety of characteristic stereotypic behaviors in adult rats. In the current study, rats received quinpirole HCl (1 mg/kg/day) or saline from postnatal day (PD) 1 to PD 11 and went otherwise untreated until adulthood (PD 60). In Experiment 1, cognitive performance was assessed on the standard and matching-to-place versions of the Morris water task (MWT). In Experiment 2, skilled motor performance was assessed on the Whishaw reaching task and locomotor activity was also analyzed. We found that ontogenetically quinpirole-treated rats displayed a deficit on the probe trial given at the end of training of the standard version of the MWT but that there were no significant differences from control on the matching-to-place task. Additionally, rats treated in ontogeny with quinpirole showed significant enhancement in reaching accuracy on the Whishaw reaching task as well as increased locomotor activity relative to saline controls. These findings demonstrate that ontogenetic quinpirole treatments produce cognitive deficits, enhanced skilled reaching and hyperlocomotion. The behavioral changes produced by ontogenetic quinpirole treatment are consistent with dopaminergic hyperfunction, and possible mechanisms are discussed.

Neonatal quinpirole (dopamine D(2)/D(3) agonist) treatment to rats has been shown to increase dopamine D(2) receptor sensitivity throughout the animal's lifetime. Male and female Sprague-Dawley rats were neonatalally treated with quinpirole (1 mg/kg) from postnatal days (P) 1-21 and raised to adulthood. Beginning on P62, rats were administered the atypical antipsychotic olanzapine (2.5 mg/kg) twice daily for 28 days. Starting 1 day after the end of olanzapine treatment, animals were behaviorally tested on the place and match-to-place version of the Morris water maze (MWM) over seven consecutive days, and a yawning behavioral test was also performed to test for sensitivity of the D(2) receptor 1 day following MWM testing. Similar to results from a past study, olanzapine alleviated cognitive impairment on the MWM place version and increases in yawning produced by neonatal quinpirole treatment. Brain tissue analyses showed that neonatal quinpirole treatment resulted in a significant decrease of hipppocampal ChAT and BDNF RNA expression that were unaffected by adulthood olanzapine treatment, although adulthood olanzapine treatment produced a significant increase in cerebellar ChAT RNA expression. There were no significant effects of drug treatment on NGF RNA expression in any brain area. These results show that neonatal quinpirole treatment produced significant decreases of protein RNA expression that is specific to the hippocampus. Although olanzapine alleviated cognitive deficits produced by neonatal quinpirole treatment, it did not affect expression of proteins known to be important in cognitive performance.

As principais incapacidades funcionais que se observam após o Acidente Vascular Encefálico (AVE), sob o ponto de vista clínico, decorrem da hemiparesia, da incoordenação, da hipertonia espástica dos membros superior e inferior contralaterais à lesão e da fraqueza ipsilateral e contralateral ao hemisfério lesado. Evidências sugerem que o aprendizado e a realização de tarefas motoras de habilidade podem induzir mudanças comportamentais e neurofisiológicas, o que ocorre tanto em animais intactos quanto naqueles submetidos às lesões do SNC. Nesse sentido, alguns trabalhos evidenciam a participação da atividade dendrítica, observada pelo aumento da imunorreatividade a MAP2, induzida por experiências comportamentais. Sendo assim, o objetivo do presente estudo foi avaliar o desempenho motor e a expressão da MAP2 no córtex motor primário, em ratos sham ou submetidos à HIC e aos treinamentos de habilidade do alcance (TH) ou ao treinamento de não-habilidade (TNH). Para tanto, ratos Wistar adultos foram inicialmente adaptados às diferentes tarefas motoras empregadas ao longo de três semanas, sendo, posteriormente, submetidos à cirurgia de indução da hemorragia intracerebral (HIC) por meio da administração intra-estriatal de colagenase tipo IV ou de veículo (animais Sham) (S). Em seguida, os animais dos grupos S_TH e HIC_TH foram submetidos ao treinamento da tarefa de habilidade do alcance e preensão, os animais dos grupos S_TNH e HIC_TNH foram submetidos ao treinamento da tarefa de não-habilidade e os animais S_ST e HIC_ST não receberam nenhum tipo de treinamento durante 4 semanas. Ao longo desse período, os animais foram testados pelo teste do Staircase quanto ao desempenho motor ao final da 2° e 4° semana de treinamento. Encerrado o período de treinamento, os animais foram profundamente anestesiados, perfundidos e tiveram seus encéfalos processados para a análise imunoistoquímica. Os resultados mostram que a realização da tarefa de habilidade do alcance e preensão foi capaz de aumentar a imunorreatividade da MAP2 no córtex motor primário (M1) em ambos os hemisférios, tanto em animais lesados quanto em animais não-lesados. Além disso, os animais HIC e HIC_TNH apresentaram também aumento da imunorreatividade à MAP2 em ambos os hemisférios. Porém, apenas os animais HIC_TH apresentaram recuperação funcional dos movimentos do membro anterior afetado, avaliados pelo teste comportamental. Concluindo, o presente estudo demonstra que o treino de habilidade induz plasticidade dendrítica no M1 em condições normalidade e lesão e, como estratégia de reabilitação, mostra-se superior ao treino de não-habilidade, na recuperação funcional do membro anterior após a HIC experimental. / Under clinical view, the main functional impairment observed after stroke is resulting from the hemiparesis, incoordination, spastic hypertonia and from ipsilateral and contralateral weakness . Evidences suggest that learning and achievement of motor tasks ability may induce behavioral and neurophysiological changes, which occur in both intact and injured animals. Accordingly, some studies reveal the participation of dendritic activity, observed by increasing the immunoreactivity to MAP2, induced by behavioral experiences. Thus, the aim of this study was to evaluate the motor performance and the expression of MAP2 in primary motor cortex (M1), in rats submitted or not to the IHC and rehabilitation using skilled (SK) or unskilled (US) training. Animals were initially adapted to different motor tasks employed over three weeks, and, subsequently, submitted to surgery for the induction of intracerebral hemorrhage (IHC) by means of administration of bacterial collagenase type IV or vehicle (animals Sham) (S) into the striatum . Then, animals in groups S_SK and IHC_SK were submitted to the training skilled forelimb reaching, animals in groups S_US and IHC_US were submitted to the training unskilled and animals S and HIC received no type of training during 4 weeks. Throughout that period, the reaching ability was tested using the Staircase test at the end of 2nd and 4thweek of training. At the end of the rehabilitation period, animals were deeply anesthetized, perfused and the immunohistochemistry was processed. Results show that the achievement of the task skilled forelimb reaching was able to increase the MAP2 immunoreactivity in primary motor cortex (M1) in both hemispheres, both in injured animals as in intact animals. In addition, animals form IHC and IHC_US groups also presented increased immunoreactivity to MAP2 in both cerebral hemispheres. However, only IHC_SK animals presented functional recovery of movements of the forelimb, evaluated by test behavioral. In conclusion, this study shows that training of skills tasks can induce modifications in M1 under conditions of normality and lesion and, as a strategy of rehabilitation, induced higher plasticity than the unskilled training correlated with functional recovery of the forelimb after IHC experimental.

As principais incapacidades funcionais que se observam após o Acidente Vascular Encefálico (AVE), sob o ponto de vista clínico, decorrem da hemiparesia, da incoordenação, da hipertonia espástica dos membros superior e inferior contralaterais à lesão e da fraqueza ipsilateral e contralateral ao hemisfério lesado. Evidências sugerem que o aprendizado e a realização de tarefas motoras de habilidade podem induzir mudanças comportamentais e neurofisiológicas, o que ocorre tanto em animais intactos quanto naqueles submetidos às lesões do SNC. Nesse sentido, alguns trabalhos evidenciam a participação da atividade dendrítica, observada pelo aumento da imunorreatividade a MAP2, induzida por experiências comportamentais. Sendo assim, o objetivo do presente estudo foi avaliar o desempenho motor e a expressão da MAP2 no córtex motor primário, em ratos sham ou submetidos à HIC e aos treinamentos de habilidade do alcance (TH) ou ao treinamento de não-habilidade (TNH). Para tanto, ratos Wistar adultos foram inicialmente adaptados às diferentes tarefas motoras empregadas ao longo de três semanas, sendo, posteriormente, submetidos à cirurgia de indução da hemorragia intracerebral (HIC) por meio da administração intra-estriatal de colagenase tipo IV ou de veículo (animais Sham) (S). Em seguida, os animais dos grupos S_TH e HIC_TH foram submetidos ao treinamento da tarefa de habilidade do alcance e preensão, os animais dos grupos S_TNH e HIC_TNH foram submetidos ao treinamento da tarefa de não-habilidade e os animais S_ST e HIC_ST não receberam nenhum tipo de treinamento durante 4 semanas. Ao longo desse período, os animais foram testados pelo teste do Staircase quanto ao desempenho motor ao final da 2° e 4° semana de treinamento. Encerrado o período de treinamento, os animais foram profundamente anestesiados, perfundidos e tiveram seus encéfalos processados para a análise imunoistoquímica. Os resultados mostram que a realização da tarefa de habilidade do alcance e preensão foi capaz de aumentar a imunorreatividade da MAP2 no córtex motor primário (M1) em ambos os hemisférios, tanto em animais lesados quanto em animais não-lesados. Além disso, os animais HIC e HIC_TNH apresentaram também aumento da imunorreatividade à MAP2 em ambos os hemisférios. Porém, apenas os animais HIC_TH apresentaram recuperação funcional dos movimentos do membro anterior afetado, avaliados pelo teste comportamental. Concluindo, o presente estudo demonstra que o treino de habilidade induz plasticidade dendrítica no M1 em condições normalidade e lesão e, como estratégia de reabilitação, mostra-se superior ao treino de não-habilidade, na recuperação funcional do membro anterior após a HIC experimental. / Under clinical view, the main functional impairment observed after stroke is resulting from the hemiparesis, incoordination, spastic hypertonia and from ipsilateral and contralateral weakness . Evidences suggest that learning and achievement of motor tasks ability may induce behavioral and neurophysiological changes, which occur in both intact and injured animals. Accordingly, some studies reveal the participation of dendritic activity, observed by increasing the immunoreactivity to MAP2, induced by behavioral experiences. Thus, the aim of this study was to evaluate the motor performance and the expression of MAP2 in primary motor cortex (M1), in rats submitted or not to the IHC and rehabilitation using skilled (SK) or unskilled (US) training. Animals were initially adapted to different motor tasks employed over three weeks, and, subsequently, submitted to surgery for the induction of intracerebral hemorrhage (IHC) by means of administration of bacterial collagenase type IV or vehicle (animals Sham) (S) into the striatum . Then, animals in groups S_SK and IHC_SK were submitted to the training skilled forelimb reaching, animals in groups S_US and IHC_US were submitted to the training unskilled and animals S and HIC received no type of training during 4 weeks. Throughout that period, the reaching ability was tested using the Staircase test at the end of 2nd and 4thweek of training. At the end of the rehabilitation period, animals were deeply anesthetized, perfused and the immunohistochemistry was processed. Results show that the achievement of the task skilled forelimb reaching was able to increase the MAP2 immunoreactivity in primary motor cortex (M1) in both hemispheres, both in injured animals as in intact animals. In addition, animals form IHC and IHC_US groups also presented increased immunoreactivity to MAP2 in both cerebral hemispheres. However, only IHC_SK animals presented functional recovery of movements of the forelimb, evaluated by test behavioral. In conclusion, this study shows that training of skills tasks can induce modifications in M1 under conditions of normality and lesion and, as a strategy of rehabilitation, induced higher plasticity than the unskilled training correlated with functional recovery of the forelimb after IHC experimental.

As principais incapacidades funcionais que se observam após o Acidente Vascular Encefálico (AVE), sob o ponto de vista clínico, decorrem da hemiparesia, da incoordenação, da hipertonia espástica dos membros superior e inferior contralaterais à lesão e da fraqueza ipsilateral e contralateral ao hemisfério lesado. Evidências sugerem que o aprendizado e a realização de tarefas motoras de habilidade podem induzir mudanças comportamentais e neurofisiológicas, o que ocorre tanto em animais intactos quanto naqueles submetidos às lesões do SNC. Nesse sentido, alguns trabalhos evidenciam a participação da atividade dendrítica, observada pelo aumento da imunorreatividade a MAP2, induzida por experiências comportamentais. Sendo assim, o objetivo do presente estudo foi avaliar o desempenho motor e a expressão da MAP2 no córtex motor primário, em ratos sham ou submetidos à HIC e aos treinamentos de habilidade do alcance (TH) ou ao treinamento de não-habilidade (TNH). Para tanto, ratos Wistar adultos foram inicialmente adaptados às diferentes tarefas motoras empregadas ao longo de três semanas, sendo, posteriormente, submetidos à cirurgia de indução da hemorragia intracerebral (HIC) por meio da administração intra-estriatal de colagenase tipo IV ou de veículo (animais Sham) (S). Em seguida, os animais dos grupos S_TH e HIC_TH foram submetidos ao treinamento da tarefa de habilidade do alcance e preensão, os animais dos grupos S_TNH e HIC_TNH foram submetidos ao treinamento da tarefa de não-habilidade e os animais S_ST e HIC_ST não receberam nenhum tipo de treinamento durante 4 semanas. Ao longo desse período, os animais foram testados pelo teste do Staircase quanto ao desempenho motor ao final da 2° e 4° semana de treinamento. Encerrado o período de treinamento, os animais foram profundamente anestesiados, perfundidos e tiveram seus encéfalos processados para a análise imunoistoquímica. Os resultados mostram que a realização da tarefa de habilidade do alcance e preensão foi capaz de aumentar a imunorreatividade da MAP2 no córtex motor primário (M1) em ambos os hemisférios, tanto em animais lesados quanto em animais não-lesados. Além disso, os animais HIC e HIC_TNH apresentaram também aumento da imunorreatividade à MAP2 em ambos os hemisférios. Porém, apenas os animais HIC_TH apresentaram recuperação funcional dos movimentos do membro anterior afetado, avaliados pelo teste comportamental. Concluindo, o presente estudo demonstra que o treino de habilidade induz plasticidade dendrítica no M1 em condições normalidade e lesão e, como estratégia de reabilitação, mostra-se superior ao treino de não-habilidade, na recuperação funcional do membro anterior após a HIC experimental. / Under clinical view, the main functional impairment observed after stroke is resulting from the hemiparesis, incoordination, spastic hypertonia and from ipsilateral and contralateral weakness . Evidences suggest that learning and achievement of motor tasks ability may induce behavioral and neurophysiological changes, which occur in both intact and injured animals. Accordingly, some studies reveal the participation of dendritic activity, observed by increasing the immunoreactivity to MAP2, induced by behavioral experiences. Thus, the aim of this study was to evaluate the motor performance and the expression of MAP2 in primary motor cortex (M1), in rats submitted or not to the IHC and rehabilitation using skilled (SK) or unskilled (US) training. Animals were initially adapted to different motor tasks employed over three weeks, and, subsequently, submitted to surgery for the induction of intracerebral hemorrhage (IHC) by means of administration of bacterial collagenase type IV or vehicle (animals Sham) (S) into the striatum . Then, animals in groups S_SK and IHC_SK were submitted to the training skilled forelimb reaching, animals in groups S_US and IHC_US were submitted to the training unskilled and animals S and HIC received no type of training during 4 weeks. Throughout that period, the reaching ability was tested using the Staircase test at the end of 2nd and 4thweek of training. At the end of the rehabilitation period, animals were deeply anesthetized, perfused and the immunohistochemistry was processed. Results show that the achievement of the task skilled forelimb reaching was able to increase the MAP2 immunoreactivity in primary motor cortex (M1) in both hemispheres, both in injured animals as in intact animals. In addition, animals form IHC and IHC_US groups also presented increased immunoreactivity to MAP2 in both cerebral hemispheres. However, only IHC_SK animals presented functional recovery of movements of the forelimb, evaluated by test behavioral. In conclusion, this study shows that training of skills tasks can induce modifications in M1 under conditions of normality and lesion and, as a strategy of rehabilitation, induced higher plasticity than the unskilled training correlated with functional recovery of the forelimb after IHC experimental.