Âmbar Southwell configurar o aparelho e treinou os autores no seu uso. Financiamento foi fornecido por uma concessão do NINDS para Paul Patterson.

1. Este protocolo é baseado na Aqueles de Southwell et. al. (2009) e Carter et. al. (2001). <ol><li> O aparelho consiste de um feixe de raios metros com uma superfície plana de 12 mm ou 6 mm de largura) de descanso de 50 cm acima do tampo da mesa em dois pólos. A caixa preta é colocada no final do feixe como o ponto de chegada. (Plywood e outros tipos de madeira para as vigas, postes, e caixa pode ser encontrada em lojas de ferragens). Material de nidificação de gaiolas casa é colocada na caixa preta para atrair o mouse para o ponto de chegada. Uma lâmpada (com 60 lâmpada watt) é usado para brilhar a luz acima do ponto inicial e serve como um estímulo aversivo. O tempo para atravessar o centro de 80 centímetros é medido por dois detectores de movimento (fabricado pela Visopia): uma em 0 cm, que inicia um timer e um de 80 cm, que pára o timer. A rede de nylon (que pode ser obtido a partir de uma loja de tecidos local) é esticada abaixo da viga, ~ 7,5 cm acima do tampo da mesa, para amortecer qualquer queda. Uma câmera de vídeo é definida em um tripé para registrar a performance. O aparelho utilizado nesta demonstração foi construído no local. </li><li> Um grupo de pelo menos 10 ratos para cada condição é recomendado para este teste. Os ratos são alojadas em grupos, quando possível, em uma luz 12 horas (sete horas - sete horas) / ciclo escuro e aclimatados a suas gaiolas na instalação comportamentais por pelo menos duas semanas antes do início do teste. Cerca de 10 min antes do treino / teste, os ratos podem ser transportados para a sala contendo o aparelho de feixe. </li><li> Em dias de treino, cada rato cruza o feixe de 12 mm 3 vezes e em seguida, o feixe de 6 milímetros 3 vezes. Vigas de larguras mais ampla (ou seja, 28 mm) pode ser usado para os ratos que não são capazes de atravessar os estreitos ou para ratos maiores. Os ratos são colocados em uma extremidade de um feixe eo tempo necessário para atravessar para o caixa de escapar do outro lado (80 cm de distância) é medido pelos detectores de movimento. O timer é iniciado pelo nariz do mouse entrarem no centro 80 cm, e parado quando o animal atinge o final da década de 80 cm. O resto camundongos por 10 min em suas gaiolas de casa entre as sessões de formação sobre as duas vigas. Durante este período intertrial, outra ratos 3-4 podem ser treinados para fazer uso eficiente do tempo. Camundongos C57BL / 6 tipicamente atravessar o feixe com uma quantidade mínima de parar ou de avaria. No entanto, quando o fazem parar, cheirar ou olhar em volta sem proceder a frente, o investigador deve incentivar o mouse para continuar avançando pelo estímulo, cutucando, ou empurrando-o por trás com os dedos enluvados. Uma vez que os ratos estão no cofre, eles são autorizados algum tempo (~ 15 segundos) para descansar lá antes do julgamento que vem. Após cada sessão de treinamento, os ratos são devolvidos diretamente para suas gaiolas de casa e levado de volta para sua sala de habitação. De nota, os ratos podem parar mais, se eles são mais treinados e tornar-se muito familiarizados com a tarefa e aparelhos. Por outro lado, a formação pode ser continuado por dias adicionais se os ratos ainda não atravessar o feixe inteiro com êxito após dois dias de treinamento. Se os tratamentos são aplicados para os ratos, uma medida de referência de desempenho deve ser obtido antes do tratamento. </li><li> No dia do teste, os tempos para atravessar cada feixe são gravadas. Dois ensaios bem sucedidos em que o mouse não parar na trave são em média. Gravações de vídeo pode ser usado para análise mais fina de escorregamento e outros déficits motores observável. Uma forma alternativa para avaliar a pé por um raio é 1,2 neurológicos sistema de pontuação, que avalia a incapacidade de atravessar o feixe como uma pontuação de &quot;1&quot; e a capacidade de atravessar a viga, normalmente com as duas patas na superfície do feixe e menos de duas mudas pé como um &quot;7&quot;. Um deslizamento é definido como o pé saindo do topo da viga. Neste sistema de indexação, uma pontuação de &quot;3&quot; corresponderia a atravessar a viga, arrastando membros. Um animal que marca 1 em dois ensaios não é reanalisada. Descrição do desempenho na trave relacionadas com outros escores pode ser encontrado em Feeney et al., (1982) e Carter et al., (2001). </li><li> As vigas e caixa são limpos de fezes de rato e enxugou com toalhas embebidas com etanol 70% e, em seguida, o feixe de água antes de seguir é colocado sobre o aparelho. </li></ol> 2. Resultados representativos: <img src="/files/ftp_upload/2376/2376fig1.jpg" alt="Figura 1" /> Figura 1. Os tempos de cruz de C57BL6 masculino e feminino em vigas 6mm e 12mm. Um grupo de 9 a 11 semanas de idade camundongos WT C57BL/6-NCRL foram testados em ambos os feixes, após dois dias de treinamento. Vezes a média para atravessar o feixe de 12 mm foram de 4,6 ± 0,4 seg para o sexo masculino (n = 14) e 3,3 ± 0,3 s para o sexo feminino (n = 11). Vezes na trave de 6 mm foram de 6,8 ± 0,7 s para o sexo masculino (n = 13) e 5,9 ± 0,5 s para o sexo feminino (n = 10) (os erros são relatados como SEM).

<ol>
	<li>Feeney, D. M., Gonzalez, A., Law, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amphetamine,+haloperidol,+and+experience+interact+to+affect+rate+of+recovery+after+motor+cortex+injury.">Amphetamine, haloperidol, and experience interact to affect rate of recovery after motor cortex injury.</a> Science. 217, 855-857 (1982).</li><li>Carter, R. J., Morton, J., Dunnett, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+coordination+and+balance+in+rodents.">Motor coordination and balance in rodents.</a> Curr Protoc Neurosci. Chapter 8, (2001).</li><li>Wallace, J. E., Krauter, E. E., Campbell, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+and+reflexive+behavior+in+the+aging+rat.">Motor and reflexive behavior in the aging rat.</a> J Gerontol. 35, 364-370 (1980).</li><li>Brooks, S. P., Dunnett, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tests+to+assess+motor+phenotype+in+mice:+a+user's+guide.">Tests to assess motor phenotype in mice: a user's guide.</a> Nat Rev Neurosci. 10, 519-529 (2009).</li><li>Fox, G. B., Fan, L., Levasseur, R. A., Faden, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+sensory/motor+and+cognitive+deficits+with+neuronal+apoptosis+following+controlled+cortical+impact+brain+injury+in+the+mouse.">Sustained sensory/motor and cognitive deficits with neuronal apoptosis following controlled cortical impact brain injury in the mouse.</a> J Neurotrauma. 15, 599-614 (1998).</li><li>Buccafusco, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Methods of behavioral analysis in neuroscience. , (2009).</li><li>Southwell, A. L., Ko, J., Patterson, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrabody+gene+therapy+ameliorates+motor,+cognitive,+and+neuropathological+symptoms+in+multiple+mouse+models+of+Huntington's+disease.">Intrabody gene therapy ameliorates motor, cognitive, and neuropathological symptoms in multiple mouse models of Huntington's disease.</a> J Neurosci. 29, 13589-13602 (2009).</li><li>Carter, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+progressive+motor+deficits+in+mice+transgenic+for+the+human+Huntington's+disease+mutation.">Characterization of progressive motor deficits in mice transgenic for the human Huntington's disease mutation.</a> J Neurosci. 19, 3248-3257 (1999).</li><li>Gulinello, M., Chen, F., Dobrenis, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+deficits+in+motor+coordination+and+cognitive+dysfunction+in+a+mouse+model+of+the+neurodegenerative+lysosomal+storage+disorder,+Sandhoff+disease.">Early deficits in motor coordination and cognitive dysfunction in a mouse model of the neurodegenerative lysosomal storage disorder, Sandhoff disease.</a> Behav Brain Res. 193, 315-319 (2008).</li><li>Stanley, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mouse+beam+walking+assay+offers+improved+sensitivity+over+the+mouse+rotarod+in+determining+motor+coordination+deficits+induced+by+benzodiazepines.">The mouse beam walking assay offers improved sensitivity over the mouse rotarod in determining motor coordination deficits induced by benzodiazepines.</a> J Psychopharmacol. 19, 221-227 (2005).</li></ol>

Experimentos em animais foram realizados de acordo com as diretrizes e regulamentos estabelecidos pela Animal Care Caltech Institucional e Comitê de Uso (IACUC).

Desempenho na trave de equilíbrio é uma medida útil de coordenação fina e equilíbrio e foi validado por trabalhos anteriores. O teste de feixe pode detectar déficits motores devido à idade, nervoso central syst `lesões, e manipulações genéticas e farmacológicas em roedores jovens e idosos 2,3,4. Camundongos com lesões impacto cortical freqüentemente apresentam contralateral escorregar na trave 5,6. O teste de feixe também tem sido utilizada para diferenciar as habilidades motoras de tipo selvagem (WT) e doença de Huntington modelos (HD) do mouse ao longo do tempo e para avaliar melhorias em camundongos HD com tratamento 7,8. Outro modelo de rato genético onde déficits motores foram detectados na trave incluem um modelo do rato de Sandhoff doença, um distúrbio de depósito lisossômico 9. Na detecção de déficits de coordenação motora induzida por benzodiazepínicos, o teste de viga foi mais sensível que o Rotarod. Stanley (2005) mostrou que apenas 30% de ocupação dos receptores GABA (A) pelo diazepam ou lorazepam foi necessária a fim de observar déficit motor na trave em comparação com mais de 70% de ocupação dos receptores de déficits na Rotarod 10. Vigas de larguras mais estreitas podem ser usados ​​para detectar diferenças ainda mais sutis. Neste estudo, testamos uma coorte de 9 a 11 semanas de idade camundongos WT C57BL/6-NCRL sobre a 12 mm e 6 vigas mm. Vezes a média para atravessar o feixe de 12 mm foram de 4,6 ± 0,4 seg para o sexo masculino (n = 14) e 3,3 ± 0,3 s para o sexo feminino (n = 11). Vezes na trave de 6 mm foram de 6,8 ± 0,7 s para o sexo masculino (n = 13) e 5,9 ± 0,5 s para o sexo feminino (n = 10) (errorsare relatado como SEM). Houve deslizamentos poucos e não cai por esses ratos sobre essas vigas. Em geneticamente ou farmacologicamente animais manipulados, o escorrega e cai pode se tornar mais freqüentes e podem ser quantificados. Camundongos prejudicada pode se agarrar para o lado da viga, o que aumenta seu tempo de atravessar. O nível de atividade pode variar, dependendo da idade e linhagem de camundongos. Camundongos C57BL / 6 são geralmente animais ativos 6. Modificações deste teste que pode incentivar os ratos para avançar incluem o uso de um feixe inclinado ao invés de um horizontal um 2,4. Os ratos também podem ser colocados na plataforma segura por alguns segundos para aumentar a vontade de alcançar a meta 2. Fatores de confusão para a avaliação da coordenação motora utilizando a trave de equilíbrio incluem motivação, e os ratos que são overtraining pode tornar-se aborrecido e não cooperativos. Se déficits motores são encontrados, outros testes, incluindo a análise da marcha pegada locomotor, malha de escalada, natação, pólo de escalada, eo teste de escada pode ser usada para avaliar a 2,4 déficits.

<ul><li> Feixes: 1-m tiras de madeira liso com quadrados pequenos, médios e grandes (ie, 6, 12 e 28 mm de largura) seções transversais </li><li> Apoio estão para realizar-se a seção início do feixe elevado (~ 1,5 cm-cross-section, 50 cm de altura) </li><li> Caixa de meta (20 cm de cada lado) montado em um suporte (3 centímetros de seção transversal, 50 cm de altura) </li><li> Hammock-~ 7,5 cm acima de mesa </li><li> 60-watt (W) lâmpada de mesa </li><li> Câmera de vídeo, tripé, e fitas de vídeo em branco </li><li> Etanol, toalhas de papel, materiais de limpeza </li></ul>

assessment of motor balance and coordination in mice using the balance beam

Lesão cerebral, manipulações genéticas, e tratamentos farmacológicos pode resultar em alterações das habilidades motoras em camundongos. Coordenação motora fina e equilíbrio pode ser avaliada pelo ensaio andando feixe. O objetivo deste teste é para o mouse para ficar de pé e caminhar através de um feixe estreito elevada a uma plataforma segura. Este teste acontece durante três dias consecutivos: dois dias de treinamento e um dia de testes. Desempenho na trave é quantificado através da medição do tempo que leva para o mouse para atravessar a viga eo número de deslizamentos de pata que ocorrem no processo. Aqui nós relatamos o protocolo utilizado em nosso laboratório, e os resultados representante de um grupo de camundongos C57BL / 6. Esta tarefa é particularmente útil para a detecção de déficits sutis na coordenação motora e equilíbrio que não podem ser detectados por testes motores, tais como o Rotarod.

Watch this Scientific Journal Video about Assessment of Motor Balance and Coordination in Mice using the Balance Beam at JoVE.com

Assessment of Motor Balance and Coordination in Mice using the Balance Beam

Déficits na coordenação motora fina podem ser avaliados com o teste de trave de equilíbrio. Desempenho na trave é quantificado pela velocidade com que o feixe é percorrida eo número de vezes que o mouse desliza na trave.

Avaliação do Equilíbrio e Coordenação Motor em camundongos usando o feixe de equilíbrio

Brain injury, genetic manipulations, and pharmacological treatments can result in alterations of motor skills in mice.  Fine motor coordination and ...

assessment-motor-balance-coordination-mice-using-balance

Nanyang Technological University

Research

JoVE Journal

Neuroscience

46.3K visualizações.  California Institute of Technology. Déficits na coordenação motora fina podem ser avaliados com o teste de trave de equilíbrio. Desempenho na trave é quantificado pela velocidade com que o feixe é percorrida eo número de vezes que o mouse desliza na trave.

Vídeo: Assessment of Motor Balance and Coordination in Mice using the Balance Beam

Measuring Motor Coordination in Mice

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such as emotionality or cognition can be assessed, however, it is vital to determine whether the motor capabilities of e.g. a mutant or lesioned mouse allow such an assessment. Performance on a maze task requiring strength and coordination, such as the Morris water maze, might well be impaired in a mouse by motor, rather than cognitive, impairments, so it is essential to selectively dissect the latter from the former. For example, sensorimotor impairments caused by NMDA antagonists have been shown to impair water maze performance2. 
Motor coordination has traditionally been assessed in mice and rats by the rotarod test, in which the animal is placed on a horizontal rod that rotates about its long axis; the animal must walk forwards to remain upright and not fall off. Both set speed and accelerating versions of the rotarod are available.
The other three tests described in this article (horizontal bar, static rods and parallel bars) all measure coordination on static apparatus. The horizontal bar also requires strength for adequate performance, particularly of the forelimbs as the mouse initially grips the bar just with the front paws. 
Adult rats do not perform well on tests such as the static rods and parallel bars (personal observations); they appear less well coordinated than mice. I have only tested male rats, however, and male mice seem generally less well coordinated than females. Mice appear to have a higher strength:weight ratio than rats; the Latin name, Mus musculus, seems entirely appropriate. The rotarod, the variations of the foot fault test12 or the Catwalk (Noldus)15 apparatus are generally used to assess motor coordination in rats.

Protocols are presented for two established motor coordination tasks, the accelerating rotarod and horizontal bar, also two tests developed in Oxford recently, the static rods and parallel bars. These tests can detect motor impairments potentially of interest in their own right, as well as being possible variables in tests of other areas of behavior.

Medindo Coordenação Motora em Ratos

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such ...

Neurociência

Assessment of Morphine-induced Hyperalgesia and Analgesic Tolerance in Mice Using Thermal and Mechanical Nociceptive Modalities

Opioid-induced hyperalgesia and tolerance severely impact the clinical efficacy of opiates as pain relievers in animals and humans. The molecular mechanisms underlying both phenomena are not well understood and their elucidation should benefit from the study of animal models and from the design of appropriate experimental protocols. 
We describe here a methodological approach for inducing, recording and quantifying morphine-induced hyperalgesia as well as for evidencing analgesic tolerance, using the tail-immersion and tail pressure tests in wild-type mice. As shown in the video, the protocol is divided into five sequential steps. Handling and habituation phases allow a safe determination of the basal nociceptive response of the animals. Chronic morphine administration induces significant hyperalgesia as shown by an increase in both thermal and mechanical sensitivity, whereas the comparison of analgesia time-courses after acute or repeated morphine treatment clearly indicates the development of tolerance manifested by a decline in analgesic response amplitude. This protocol may be similarly adapted to genetically modified mice in order to evaluate the role of individual genes in the modulation of nociception and morphine analgesia. It also provides a model system to investigate the effectiveness of potential therapeutic agents to improve opiate analgesic efficacy.

We describe a protocol to examine the development of opioid-induced hyperalgesia and tolerance in mice. Based on the measurement of thermal and mechanical nociceptive responses of na&#239;ve and morphine-treated animals, it allows to quantify the increase in pain sensitivity (hyperalgesia) and decrease in analgesia (tolerance) associated with chronic opiate administration.

Avaliação da hiperalgesia induzida por morfina e analgésicos Tolerância em ratos usando térmicas e mecânicas nociceptivos Modalidades

Opioid-induced hyperalgesia and tolerance severely impact the clinical efficacy of opiates as pain relievers in animals and humans. The molecular ...

Assessment of Vascular Regeneration in the CNS Using the Mouse Retina

The rodent retina is perhaps the most accessible mammalian system in which to investigate neurovascular interplay within the central nervous system (CNS). It is increasingly being recognized that several neurodegenerative diseases such as Alzheimer&#8217;s, multiple sclerosis, and amyotrophic lateral sclerosis present elements of vascular compromise. In addition, the most prominent causes of blindness in pediatric and working age populations (retinopathy of prematurity and diabetic retinopathy, respectively) are characterized by vascular degeneration and failure of physiological vascular regrowth. The aim of this technical paper is to provide a detailed protocol to study CNS vascular regeneration in the retina. The method can be employed to elucidate molecular mechanisms that lead to failure of vascular growth after ischemic injury. In addition, potential therapeutic modalities to accelerate and restore healthy vascular plexuses can be explored. Findings obtained using the described approach may provide therapeutic avenues for ischemic retinopathies such as that of diabetes or prematurity and possibly benefit other vascular disorders of the CNS.

The rodent retina has long been recognized as an accessible window to the brain. In this technical paper we provide a protocol that employs the mouse model of oxygen-induced retinopathy to study the mechanisms that lead to failure of vascular regeneration within the central nervous system after ischemic injury. The described system can also be harnessed to explore strategies to promote regrowth of functional blood vessels within the retina and CNS.

Assessment of Vascular regeneração no SNC Usando o rato Retina

The rodent retina is perhaps the most accessible mammalian system in which to investigate neurovascular interplay within the central nervous system (CNS). ...

The Neuromuscular Junction: Measuring Synapse Size, Fragmentation and Changes in Synaptic Protein Density Using Confocal Fluorescence Microscopy

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. Structural changes at the NMJ can result in neurotransmission failure, resulting in weakness, atrophy and even death of the muscle fiber. Many studies have investigated how genetic modifications or disease can alter the structure of the mouse NMJ. Unfortunately, it can be difficult to directly compare findings from these studies because they often employed different parameters and analytical methods. Three protocols are described here. The first uses maximum intensity projection confocal images to measure the area of acetylcholine receptor (AChR)-rich postsynaptic membrane domains at the endplate and the area of synaptic vesicle staining in the overlying presynaptic nerve terminal. The second protocol compares the relative intensities of immunostaining for synaptic proteins in the postsynaptic membrane. The third protocol uses Fluorescence Resonance Energy Transfer (FRET) to detect changes in the packing of postsynaptic AChRs at the endplate. The protocols have been developed and refined over a series of studies. Factors that influence the quality and consistency of results are discussed and normative data are provided for NMJs in healthy young adult mice.

The neuromuscular junction (NMJ) is altered in a variety of conditions that can sometimes culminate in synaptic failure. This report describes fluorescence microscope-based methods to quantify such structural changes.

A Junção Neuromuscular: Synapse Medir o tamanho, Mudanças de fragmentação e em Synaptic Density proteína utilizando microscopia de fluorescência confocal

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. ...

Assessment of Dendritic Arborization in the Dentate Gyrus of the Hippocampal Region in Mice

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity and extent of dendritic arborization correlates well with the synaptic plasticity in these cells. A reliable method for assessment of dendritic arborization is needed to characterize the deleterious effects of neurological disorders on these structures and to determine the effects of therapeutic interventions. However, quantification of these structures has proven to be a formidable task given their complex and dynamic nature. Fortunately, sophisticated imaging techniques can be paired with conventional staining methods to assess the state of dendritic arborization, providing a more reliable and expeditious means of assessment. Below is an example of how these imaging techniques were paired with staining methods to characterize the dendritic arborization in wild type mice. These complementary imaging methods can be used to qualitatively and quantitatively assess dendritic arborization that span a rather wide area within the hippocampal region.

We describe two methods for visualization and quantification of dendritic arborization in the hippocampus of mouse models: real-time and extended depth of field imaging. While the former method allows sophisticated topographical tracing and quantification of the extent of branching, the latter allows speedy visualization of the dendritic tree.

Avaliação de dendríticas Arborização no giro dentado do hipocampo em Ratos Região

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity ...

A Human-machine-interface Integrating Low-cost Sensors with a Neuromuscular Electrical Stimulation System for Post-stroke Balance Rehabilitation

A stroke is caused when an artery carrying blood from heart to an area in the brain bursts or a clot obstructs the blood flow to brain thereby preventing delivery of oxygen and nutrients. About half of the stroke survivors are left with some degree of disability. Innovative methodologies for restorative neurorehabilitation are urgently required to reduce long-term disability. The ability of the nervous system to reorganize its structure, function and connections as a response to intrinsic or extrinsic stimuli is called neuroplasticity. Neuroplasticity is involved in post-stroke functional disturbances, but also in rehabilitation. Beneficial neuroplastic changes may be facilitated with non-invasive electrotherapy, such as neuromuscular electrical stimulation (NMES) and sensory electrical stimulation (SES). NMES involves coordinated electrical stimulation of motor nerves and muscles to activate them with continuous short pulses of electrical current while SES involves stimulation of sensory nerves with electrical current resulting in sensations that vary from barely perceivable to highly unpleasant. Here, active cortical participation in rehabilitation procedures may be facilitated by driving the non-invasive electrotherapy with biosignals (electromyogram (EMG), electroencephalogram (EEG), electrooculogram (EOG)) that represent simultaneous active perception and volitional effort. To achieve this in a resource-poor setting, e.g., in low- and middle-income countries, we present a low-cost human-machine-interface (HMI) by leveraging recent advances in off-the-shelf video game sensor technology. In this paper, we discuss the open-source software interface that integrates low-cost off-the-shelf sensors for visual-auditory biofeedback with non-invasive electrotherapy to assist postural control during balance rehabilitation. We demonstrate the proof-of-concept on healthy volunteers.

A novel low-cost human-machine interface for interactive post-stroke balance rehabilitation system is presented in this article. The system integrates off-the-shelf low-cost sensors towards volitionally driven electrotherapy paradigm. The proof-of-concept software interface is demonstrated on healthy volunteers.

A-máquina interface humana Integrando sensores de baixo custo com uma estimulação do sistema neuromuscular eléctrico para pós-AVC Reabilitação Balance

A stroke is caused when an artery carrying blood from heart to an area in the brain bursts or a clot obstructs the blood flow to brain thereby preventing ...

Functional and Morphological Assessment of Diaphragm Innervation by Phrenic Motor Neurons

This protocol specifically focuses on tools for assessing phrenic motor neuron (PhMN) innervation of the diaphragm at both the electrophysiological and morphological levels. Compound muscle action potential (CMAP) recording following phrenic nerve stimulation can be used to quantitatively assess functional diaphragm innervation by PhMNs of the cervical spinal cord in vivo in anesthetized rats and mice. Because CMAPs represent simultaneous recording of all myofibers of the whole hemi-diaphragm, it is useful to also examine the phenotypes of individual motor axons and myofibers at the diaphragm NMJ in order to track disease- and therapy-relevant morphological changes such as partial and complete denervation, regenerative sprouting and reinnervation. This can be accomplished via whole-mount immunohistochemistry (IHC) of the diaphragm, followed by detailed morphological assessment of individual NMJs throughout the muscle. Combining CMAPs and NMJ analysis provides a powerful approach for quantitatively studying diaphragmatic innervation in rodent models of CNS and PNS disease.

Compound muscle action potential recording quantitatively assesses functional diaphragm innervation by phrenic motor neurons. Whole-mount diaphragm immunohistochemistry assesses morphological innervation at individual neuromuscular junctions. The goal of this protocol is to demonstrate how these two powerful methodologies can be used in various rodent models of spinal cord disease.

Funcional e morfológica de Avaliação de Diafragma Innervation por frênico Neurônios Motores

This protocol specifically focuses on tools for assessing phrenic motor neuron (PhMN) innervation of the diaphragm at both the electrophysiological and ...

Assessment of Hippocampal Dendritic Complexity in Aged Mice Using the Golgi-Cox Method

Dendritic spines are the protuberances from the neuronal dendritic shafts that contain&#160; excitatory synapses. The morphological and branching variations of the neuronal dendrites within the hippocampus are implicated in cognition and memory formation. There are several approaches to Golgi staining, all of which have been useful for determining the morphological characteristics of dendritic arbors and produce a clear background. The present Golgi-Cox method, (a slight variation of the protocol that is provided with a commercial Golgi staining kit), was designed to assess how a relatively low dose of the chemotherapeutic drug 5-flurouracil (5-Fu) would affect dendritic morphology, the number of spines, and the complexity of arborization within the hippocampus. The 5-Fu significantly modulated the dendritic complexity and decreased the spine density throughout the hippocampus in a region-specific manner. The data presented show that the Golgi staining method effectively stained the mature neurons in the CA1, the CA3, and the dentate gyrus (DG) of the hippocampus. This protocol reports the details for each step so that other researchers can reliably stain tissue throughout the brain with high quality results and minimal troubleshooting.

Here we present a Golgi-Cox protocol in extensive detail. This reliable tissue stain method allows for a high-quality assessment of the cytoarchitecture in the hippocampus, and throughout the entire brain, with minimal troubleshooting.

Avaliação da complexidade dendrítica do hipocampo em ratos idosos usando o método de Golgi-Cox

Dendritic spines are the protuberances from the neuronal dendritic shafts that contain&#160; excitatory synapses. The morphological and branching ...

Retrograde Neuroanatomical Tracing of Phrenic Motor Neurons in Mice

Phrenic motor neurons are cervical motor neurons originating from C3 to C6 levels in most mammalian species. Axonal projections converge into phrenic nerves innervating the respiratory diaphragm. In spinal cord slices, phrenic motor neurons cannot be identified from other motor neurons on morphological or biochemical criteria. We provide the description of procedures for visualizing phrenic motor neuron cell bodies in mice, following intrapleural injections of cholera toxin subunit beta (CTB) conjugated to a fluorophore. This fluorescent neuroanatomical tracer has the ability to be caught up at the diaphragm neuromuscular junction, be carried retrogradely along the phrenic axons and reach the phrenic cell bodies. Two methodological approaches of intrapleural CTB delivery are compared: transdiaphragmatic versus transthoracic injections. Both approaches are successful and result in similar number of CTB-labeled phrenic motor neurons. In conclusion, these techniques can be applied to visualize or quantify the phrenic motor neurons in various experimental studies such as those focused on the diaphragm-phrenic circuitry.

Here, we describe a protocol for identifying phrenic motor neurons in mice after intrapleural delivery of fluorophore conjugated cholera toxin subunit beta. Two techniques are compared to inject the pleural cavity: transdiaphragmatic versus transthoracic approaches.

Rastreamento neuroanatômica retrógrado do frênico neurônios em ratos

Phrenic motor neurons are cervical motor neurons originating from C3 to C6 levels in most mammalian species. Axonal projections converge into phrenic ...

3D Kinematic Analysis for the Functional Evaluation in the Rat Model of Sciatic Nerve Crush Injury

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of sciatic nerve injury rodent models. In this protocol, we describe a novel kinematic analysis method that uses a three-dimensional (3D) motion capture apparatus for functional evaluations using a rat sciatic nerve crush injury model. First, the rat is familiarized with treadmill walking. Markers are then attached to the designated bone landmarks and the rat is made to walk on the treadmill at the desired speed. Meanwhile, the posterior limb movements of the rat are recorded using four cameras. Depending on the software used, marker tracings are created using both automatic and manual modes and the desired data are produced after subtle adjustments. This method of kinematic analysis, which uses a 3D motion capture apparatus, offers numerous advantages, including superior precision and accuracy. Many more parameters can be investigated during the comprehensive functional evaluations. This method has several shortcomings that require consideration: The system is expensive, can be complicated to operate, and may produce data deviations due to skin shifting. Nevertheless, kinematic analysis using a 3D motion capture apparatus is useful for performing functional anterior and posterior limb evaluations. In the future, this method may become increasingly useful for generating accurate assessments of various traumas and diseases.

We introduce a kinematic analysis method that uses a three-dimensional motion capture apparatus containing four cameras and data processing software for performing functional evaluations during fundamental research involving rodent models.

Análise Cinética 3D para a Avaliação Funcional no modelo de rat da lesão ciática de esmagamento nervoso

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of ...