O financiamento para esta pesquisa foi fornecido pelos passes Garnett e Rodney Williams Memorial Foundation e La Trobe University.

<ol>
	<li>Mauger, S. J., Shivdasani, M. N., Rathbone, G. D., Argent, R. E., Paolini, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+vivo+investigation+of+first+spike+latencies+in+the+inferior+colliculus+in+response+to+multichannel+penetrating+auditory+brainstem+implant+stimulation.">An in vivo investigation of first spike latencies in the inferior colliculus in response to multichannel penetrating auditory brainstem implant stimulation.</a> Journal of Neural Engineering. 7, 036004-036004 (2010).</li><li>Shivdasani, M. N., Mauger, S. J., Argent, R. E., Rathbone, G. D., Paolini, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inferior+colliculus+responses+to+dual-site+intralamina+stimulation+in+the+ventral+cochlear+nucleus.">Inferior colliculus responses to dual-site intralamina stimulation in the ventral cochlear nucleus.</a> J. Comp. Neurol. 518, 4226-4242 (2010).</li><li>Shivdasani, M. N., Mauger, S. J., Rathbone, G. D., Paolini, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inferior+colliculus+responses+to+multichannel+microstimulation+of+the+ventral+cochlear+nucleus:+Implications+for+auditory+brain+stem+implants.">Inferior colliculus responses to multichannel microstimulation of the ventral cochlear nucleus: Implications for auditory brain stem implants.</a> Journal of Neurophysiology. 99, 1-13 (2007).</li><li>Lenarz, M., Lim, H. H., Patrick, J. F., Anderson, D. J., Lenarz, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+Validation+of+a+Human+Prototype+Auditory+Midbrain+Implant+in+a+Guinea+Pig+Model.">Electrophysiological Validation of a Human Prototype Auditory Midbrain Implant in a Guinea Pig Model.</a> JARO. 7, 383-398 (2006).</li><li>Lim, H. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auditory+Cortical+Responses+to+Electrical+Stimulation+of+the+Inferior+Colliculus:+Implications+for+an+Auditory+Midbrain+Implant.">Auditory Cortical Responses to Electrical Stimulation of the Inferior Colliculus: Implications for an Auditory Midbrain Implant.</a> Journal of Neurophysiology. 96, 975-988 (2006).</li><li>Otto, K. J., Rousche, P. J., Kipke, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microstimulation+in+auditory+cortex+provides+a+substrate+for+detailed+behaviors.">Microstimulation in auditory cortex provides a substrate for detailed behaviors.</a> Hearing research. 210, 112-117 (2005).</li><li>Gai, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+Tones+in+Reproducible+Noise+Maskers+by+Rabbits+and+Comparison+to+Detection+by+Humans.">Detection of Tones in Reproducible Noise Maskers by Rabbits and Comparison to Detection by Humans.</a> JARO. 8, 522-538 (2007).</li><li>Miller, C. A., Woodruff, K. E., Pfingst, B. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+responses+from+guinea+pigs+with+cochlear+implants.+I.+Electrophysiological+and+psychophysical+measures.">Functional responses from guinea pigs with cochlear implants. I. Electrophysiological and psychophysical measures.</a> Hearing Research. 92, 85-99 (1995).</li><li>Beitel, R. E., Vollmer, M., Snyder, R. L., Schreiner, C. E., Leake, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+and+neurophysiological+thresholds+for+electrical+cochlear+stimulation+in+the+deaf+cat.">Behavioral and neurophysiological thresholds for electrical cochlear stimulation in the deaf cat.</a> Audiology &#38; Neurotology. 5, 31-38 (2000).</li><li>Beitel, R. E., Snyder, R. L., Schreiner, C. E., Raggio, M. W., Leake, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+cochlear+stimulation+in+the+deaf+cat:+comparisons+between+psychophysical+and+central+auditory+neuronal+thresholds.">Electrical cochlear stimulation in the deaf cat: comparisons between psychophysical and central auditory neuronal thresholds.</a> J. Neurophysiol. 83, 2145-2162 (2000).</li><li>Vollmer, M., Beitel, R. E., Snyder, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auditory+detection+and+discrimination+in+deaf+cats:+Psychophysical+and+neural+thresholds+for+intracochlear+electrical+signals.">Auditory detection and discrimination in deaf cats: Psychophysical and neural thresholds for intracochlear electrical signals.</a> Journal of Neurophysiology. 86, 2330-2343 (2001).</li><li>LeDoux, J. E., Cicchetti, P., Xagoraris, A., Romanski, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+lateral+amygdaloid+nucleus:+sensory+interface+of+the+amygdala+in+fear+conditioning.">The lateral amygdaloid nucleus: sensory interface of the amygdala in fear conditioning.</a> Journal of Neuroscience. 10, 1062-1069 (1990).</li><li>McCreery, D. B., Yuen, T. G., Agnew, W. F., Bullara, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+characterization+of+the+effects+on+neuronal+excitability+due+to+prolonged+microstimulation+with+chronically+implanted+microelectrodes.">A characterization of the effects on neuronal excitability due to prolonged microstimulation with chronically implanted microelectrodes.</a> IEEE Transactions on Biomedical Engineering. 44, 931-939 (1997).</li><li>Liu, X., McPhee, G., Seldon, H. L., Clark, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histological+and+physiological+effects+of+the+central+auditory+prosthesis:+surface+versus+penetrating+electrodes.">Histological and physiological effects of the central auditory prosthesis: surface versus penetrating electrodes.</a> Hearing Research. 114, 264-274 (1997).</li><li>Polikov, V. S., Tresco, P. A., Reichert, W. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Response+of+brain+tissue+to+chronically+implanted+neural+electrodes.">Response of brain tissue to chronically implanted neural electrodes.</a> Journal of Neuroscience Methods. 148, 1-18 (2005).</li></ol>

Dr. Antonio Paolini é um diretor de Bioengenesis Technologies Pty Ltd, que são os representantes australianos para Tucker Davis Technologies eo distribuidor australiano de Sondas NeuroNexus.

As técnicas descritas aqui fornecer um meio para testar uma gama de tarefas de discriminação rapidamente, com tempos de formação de curta duração e que permitem a automação substancial, minimizando o tempo pesquisador requeridos para processamento de dados. Utilizando dados obtidos a partir do cálculo implantado ECG, de frequência cardíaca (FC) mudança em tempos de apresentação do estímulo pode ser automatizado. Erros no ECG de processamento (como perder uma única batida de coração) podem ser facilmente identificados como elas produzem mudanças spuriously grandes em aparente da frequência cardíaca durante um período muito breve, e detecção de erros tão fácil minimiza a necessidade de demorada análise manual de dados . A técnica descrita para a implantação do dispositivo de telemetria ECG produz consistentemente gravações com pouca ou nenhuma interferência (ver Figuras 1 e 2). No entanto, variações relativamente pequenas na colocação do chumbo na região mediastino pode levar a interferência a partir de músculo próximas e em particular o ruído da respiração associada. Verificando a colocação correta dos eletrodos durante o processo de implantação, especialmente após a sutura leva no local, permite ajustes finos de posicionamento do eletrodo para minimizar interferências. O processo de implantação cérebro para o núcleo coclear é difícil. Ao utilizar apresentações de série passa-banda sonora filtrada de várias faixas de freqüência, ao invés de ruído de banda larga como tem sido utilizado 3, respostas de populações de células em locais de eletrodos podem ser rapidamente revisto em termos de frequência de resposta, em vez de o ruído menos específica resposta. A capacidade de identificar rapidamente quando um intervalo suficiente das populações-frequência ajustados celulares são acessíveis é fundamental na investigação quando a discriminação estímulo elétrico está sendo testado. Se insuficiente frequência de distribuição obtém-se, em seguida, a comparação entre a estimulação das zonas de frequência distintas não é simplesmente possível. No entanto, o processo maisde freqüência-amplitude mapeamento resposta continua a ser essencial para proporcionar detalhes sobre a amplitude de ajuste de populações de células e deve ser realizada antes de fixar o eletrodo no lugar. Na formação comportamental e ensaio, a inclusão de um estímulo não-(ou silenciosa) período entre ensaios, antes do início do estímulo, permite que a resposta ao início do estímulo inicial a ser examinado. Quando se utiliza estimulação neural eléctrico, uma resposta ao início do estímulo fornece evidência para a detecção simples do estímulo. Assim, quando não há resposta início do estímulo, mas uma resposta quando o estímulo alternância segundo é introduzido, pode deduzir-se que apenas estímulo o último pode ser detectado. Sem a inclusão de um período tão silencioso, não haveria qualquer maneira para determinar que o estímulo anterior não foi detectada. Uma limitação do presente técnica de teste comportamental é que a ausência de febre aftosa, choques entregue após a apresentação da Neuraestimulação l pode levar ao desenvolvimento de especificidade no medo condicionado, tal receio de que só é expressa quando o estímulo acústico é apresentado. Isto é, o animal pode saber que os efeitos particulares sensoriais de estimulação neural estão associados com não receber um choque nas patas. Em contraste, proporcionando pé-choques após a apresentação de estimulação neural pode levar a um medo condicionado do estímulo neural em si, distinto de quaisquer propriedades de som, como o estímulo neural pode ter. Os resultados obtidos quando excluindo um choque nas patas, após a estimulação neural certamente proporcionar mais forte evidência para estímulo neural sendo som semelhante do que aqueles que utilizam um choque nas patas, após a estimulação neural, tal como no caso anterior medo condicionado tem generalizada a partir de acústico para a estimulação neural. No entanto, o risco de desenvolvimento de especificidade para acústico, em vez de estimulação neural está presente. O uso de passa-banda filtrada ruído como estímulo acústico em vez de rajadas de tons puros, o primeiro dos quaispode refletir melhor a experiência perceptiva da estimulação neural, poderia reduzir o risco de tal especificidade em desenvolvimento. No entanto, um tal procedimento introduz a variável adicional de largura de banda do filtro, o que afectará tarefas de discriminação. Uma limitação adicional associado com qualquer implante crónica neural, tais como necessária para o teste comportamental, é a mudança na função do eléctrodo ou o tecido neural associado ao longo do tempo. Estimulação do tecido neural pode levar tanto à supressão temporária de 13 a atividade neural e alterações teciduais, como célula resposta perda de tecido cerebral 14 e direta ao eletrodo 15. A abordagem descrita para a implantação e teste comportamental fornece um meio para testar a detecção e discriminação de estímulos acústicos e eléctrico com a formação breve, ea capacidade de exercer um controlo de frequência ensaio. A técnica de utilização de mudança de RH como um measurE de medo condicionado pode ser aplicável a não teste auditivo só, mas mais geralmente a qualquer testes sensoriais em que estímulos podem ser apresentadas para qualquer período de discreta, e em que a detecção de discriminação sensorial é desejada.

<table border="1"><tbody><tr><td> Nome </td><td> Companhia </td><td> Modelo </td><td> Comentários </td></tr><tr><td> PowerLab </td><td> ADInstruments </td><td> ML880 </td><td> Registros de dados recebidos do transmissor implantado TR40 </td></tr><tr><td> SmartCtrl placa controladora </td><td> Med Associates </td><td> DIG-716B </td><td> Controla a câmara de ensaio comportamental, incluindo choque nas patas </td></tr><tr><td> Câmara de teste modular comportamental </td><td> Med Associates </td><td> ENV-009 </td><td> Tamanho da câmara de teste: 30.5cm x 39,4 centímetros </td></tr><tr><td> Gerador de estímulos aversivos </td><td> Med Associates </td><td> ENV-410B </td><td> Proporciona aversivo pé choque com metal-bar chão da gaiola através de uma unidade Solid State Scrambler </td></tr><tr><td> Atenuador programável </td><td> TDT </td><td> PA5 </td><td> Controla o amplitude de entregue sinal acústico para garantir amplitude estável através de frequências </td></tr><tr><td> O controlador dos altifalantes eletrostática </td><td> TDT </td><td> ED1 </td><td> Drives os alto-falantes eletrostáticos (EC1, ES1) </td></tr><tr><td> Free-campo Falante eletrostática </td><td> TDT </td><td> ES1 </td><td> Sons são apresentados usando este alto-falante na câmara de ensaio comportamental </td></tr><tr><td> Juntamente falante eletrostático </td><td> TDT </td><td> EC1 </td><td> Os sons são apresentados usando este alto-falante durante a cirurgia de implante neural </td></tr><tr><td> Estimulador Base Station </td><td> TDT </td><td> RX7 </td><td> Controlos entrega de estimulação neural eléctrico (usado com MS16) </td></tr><tr><td> Microstimulator </td><td> TDT </td><td> MS16 </td><td> Proporciona estimulação elétrica neural multicanal </td></tr><tr><td> Processamento de estação de base </td><td> TDT </td><td> RZ2 </td><td> Registros de atividade neural durante a implantação do cérebro (usando PZ2 pré-amplificador) </td></tr><tr><td> Pré-amplificador </td><td> TDT </td><td> PZ2-256 </td><td> 256-canal pré-amplificador de alta impedância </td></tr><tr><td> Receptor dispositivo de telemetria </td><td> Pesquisa Telemetria </td><td> TR162 </td><td> Recebe o sinal digital a partir de TR40, e converte a saída analógica amplificado </td></tr><tr><td> Dispositivo de telemetria implantável eletrocardiograma </td><td> Pesquisa Telemetria </td><td> TR40 </td><td> O dispositivo transmissor implantado, a amostragem em 2kHz </td></tr><tr><td> Multifuncional Processor </td><td> Tucker Davis Technologies (TDT) </td><td> RX6 </td><td> Usado para gerar estímulos acústicos </td></tr><tr><td> Vertex Castavaria </td><td> Dental Vertex </td><td></td><td> Acrílico dental usado para fixar o eletrodo no lugar </td></tr><tr><td> Kwik-Sil adesiva, baixa viscosidade </td><td> Instrumentos de precisão do mundo </td><td></td><td> Elastómero de silicone utilizado para revestir as hastes de eléctrodos </td></tr><tr><td> Eletrodo Multicanal </td><td> NeuroNexus </td><td> a2x16-10mm 100-500-413 </td><td> O 2 haste-32 canal-matriz extracelular eletrodo utilizado para a implantação. Os sítios de eléctrodos de ter sido activado para produzir um revestimento de óxido de irídio em preparação para a estimulação. </td></tr></tbody></table>

behavioral determination of stimulus pair discrimination of auditory acoustic and electrical stimuli using a classical conditioning and heart-rate approach

Preparações animais aguda têm sido utilizados em pesquisas prospectivamente investigar desenhos de eléctrodos de estimulação e técnicas para a integração próteses auditivas neurais, tais como implantes auditivo do tronco cerebral auditivo 1-3 e implantes mesencéfalo 4,5. Embora experiências agudas pode dar uma visão inicial para a eficácia do implante, o teste dos animais implantados cronicamente e desperto proporciona a vantagem de examinar as propriedades psicofísicos das sensações induzida utilizando dispositivos implantados 6,7. Várias técnicas, tais como recompensa baseada em condicionamento operante 6-8, evitar condicionado 9-11, ou o condicionamento do medo clássico 12 foram usados ​​para fornecer a confirmação comportamental de detecção de um atributo relevante de estímulo. Seleção de uma técnica envolve aspectos de equilibração, incluindo eficiência de tempo (muitas vezes pobres em recompensa baseados em abordagens), a capacidade de testar uma pluralidade deestímulo atributos simultaneamente (limitado na prevenção condicionado), e confiabilidade medida de estímulos repetidos (uma restrição potencial quando as medidas fisiológicas são empregados). Aqui, um método de condicionamento clássico medo comportamental é apresentado o que pode ser usado para testar simultaneamente ambos detecção de um estímulo, e discriminação entre dois estímulos. Da frequência cardíaca é utilizado como uma medida da resposta de medo, o que reduz ou elimina o requisito de demorada de codificação de vídeo para o comportamento de congelamento ou outros tais medidas (embora essas medidas podem ser incluídos para fornecer evidência convergente). Os animais foram condicionados utilizando estas técnicas em três sessões de 2 horas de condicionamento, cada uma com 48 testes de estímulo. Subsequentes sessões de ensaios 48-ensaio foram então usadas para testar para a detecção de cada estímulo em pares apresentados, e discriminação de ensaio entre os estímulos membros de cada par. Este método é apresentado na comportamentalno contexto da sua utilização em investigação auditivo protético. A implantação de dispositivos de telemetria eletrocardiograma é mostrado. Posterior implantação de eletrodos no cérebro para o núcleo coclear, guiada pelo monitoramento da resposta neural aos estímulos acústicos, ea fixação do eletrodo no local para uso crônico é também mostrado.

Watch this Scientific Journal Video about Behavioral Determination of Stimulus Pair Discrimination of Auditory Acoustic and Electrical Stimuli Using a Classical Conditioning and Heart-rate Approach at

Behavioral Determination of Stimulus Pair Discrimination of Auditory Acoustic and Electrical Stimuli Using a Classical Conditioning and Heart-rate Approach

A aplicação de um paradigma clássico medo condicionado comportamental para auditivo protético investigação em ratos é descrito. Este paradigma fornece um mecanismo para a identificação de ambos, detecção de e discriminação entre, acústica distinta e estímulos elétricos com freqüência cardíaca como uma medida de resultado.

Determinação do Comportamento da Discriminação do estímulo acústico Par de estímulos auditivos e elétrica usando um condicionamento clássico e da freqüência cardíaca abordagem

Acute animal preparations have been used in research prospectively investigating electrode designs and stimulation techniques for integration into neural ...

behavioral-determination-stimulus-pair-discrimination-auditory

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Unit 1

Frequency Response

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14.4K visualizações.  La Trobe University. A aplicação de um paradigma clássico medo condicionado comportamental para auditivo protético investigação em ratos é descrito. Este paradigma fornece um mecanismo para a identificação de ambos, detecção de e discriminação entre, acústica distinta e estímulos elétricos com freqüência cardíaca como uma medida de resultado.

Vídeo: Behavioral Determination of Stimulus Pair Discrimination of Auditory Acoustic and Electrical Stimuli Using a Classical Conditioning and Heart-rate Approach

Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees (Apis mellifera)

Honeybees (Apis mellifera) are well known for their communication and orientation skills and for their impressive learning capability1,2. Because the survival of a honeybee colony depends on the exploitation of food sources, forager bees learn and memorize variable flower sites as well as their profitability. Forager bees can be easily trained in natural settings where they forage at a feeding site and learn the related signals such as odor or color. Appetitive associative learning can also be studied under controlled conditions in the laboratory by conditioning the proboscis extension response (PER) of individually harnessed honeybees3,4. This learning paradigm enables the study of the neuronal and molecular mechanisms that underlie learning and memory formation in a simple and highly reliable way5-12. A behavioral pharmacology approach is used to study molecular mechanisms. Drugs are injected systemically to interfere with the function of specific molecules during or after learning and memory formation13-16.
Here we demonstrate how to train harnessed honeybees in PER conditioning and how to apply drugs systemically by injection into the bee flight muscle.

Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees Apis mellifera

We demonstrate how to implement a behavioral pharmacology method in an appetitive olfactory conditioning paradigm in honeybees (Apis mellifera) by systemic application of drugs. This method allows investigation of the mechanisms underlying learning and memory formation in a simple and reliable way.

Farmacologia Comportamental no condicionamento clássico da Resposta Extensão Proboscis em Abelhas ( Apis mellifera)

Honeybees (Apis mellifera) are well known for their communication and orientation skills and for their impressive learning capability1,2. Because the ...

Neurociência

A Lightweight, Headphones-based System for Manipulating Auditory Feedback in Songbirds

Experimental manipulations of sensory feedback during complex behavior have provided valuable insights into the computations underlying motor control and sensorimotor plasticity1. Consistent sensory perturbations result in compensatory changes in motor output, reflecting changes in feedforward motor control that reduce the experienced feedback error. By quantifying how different sensory feedback errors affect human behavior, prior studies have explored how visual signals are used to recalibrate arm movements2,3 and auditory feedback is used to modify speech production4-7. The strength of this approach rests on the ability to mimic naturalistic errors in behavior, allowing the experimenter to observe how experienced errors in production are used to recalibrate motor output.
Songbirds provide an excellent animal model for investigating the neural basis of sensorimotor control and plasticity8,9. The songbird brain provides a well-defined circuit in which the areas necessary for song learning are spatially separated from those required for song production, and neural recording and lesion studies have made significant advances in understanding how different brain areas contribute to vocal behavior9-12. However, the lack of a naturalistic error-correction paradigm - in which a known acoustic parameter is perturbed by the experimenter and then corrected by the songbird - has made it difficult to understand the computations underlying vocal learning or how different elements of the neural circuit contribute to the correction of vocal errors13.
The technique described here gives the experimenter precise control over auditory feedback errors in singing birds, allowing the introduction of arbitrary sensory errors that can be used to drive vocal learning. Online sound-processing equipment is used to introduce a known perturbation to the acoustics of song, and a miniaturized headphones apparatus is used to replace a songbird's natural auditory feedback with the perturbed signal in real time. We have used this paradigm to perturb the fundamental frequency (pitch) of auditory feedback in adult songbirds, providing the first demonstration that adult birds maintain vocal performance using error correction14. The present protocol can be used to implement a wide range of sensory feedback perturbations (including but not limited to pitch shifts) to investigate the computational and neurophysiological basis of vocal learning.

We describe the design and assembly of miniaturized headphones suitable for replacing a songbird&#x2019;s natural auditory feedback with a manipulated acoustic signal. Online sound processing hardware is used to manipulate song output, introduce real-time errors in auditory feedback via the headphones, and drive vocal motor learning.

Um leve, sistema de fones baseado para manipular feedback auditivo em Songbirds

Experimental manipulations of sensory feedback during complex behavior have provided valuable insights into the computations underlying motor control and ...

Functional Imaging of Auditory Cortex in Adult Cats using High-field fMRI

Current knowledge of sensory processing in the mammalian auditory system is mainly derived from electrophysiological studies in a variety of animal models, including monkeys, ferrets, bats, rodents, and cats. In order to draw suitable parallels between human and animal models of auditory function, it is important to establish a bridge between human functional imaging studies and animal electrophysiological studies. Functional magnetic resonance imaging (fMRI) is an established, minimally invasive method of measuring broad patterns of hemodynamic activity across different regions of the cerebral cortex. This technique is widely used to probe sensory function in the human brain, is a useful tool in linking studies of auditory processing in both humans and animals and has been successfully used to investigate auditory function in monkeys and rodents. The following protocol describes an experimental procedure for investigating auditory function in anesthetized adult cats by measuring stimulus-evoked hemodynamic changes in auditory cortex using fMRI. This method facilitates comparison of the hemodynamic responses across different models of auditory function thus leading to a better understanding of species-independent features of the mammalian auditory cortex.

Functional studies of the auditory system in mammals have traditionally been conducted using spatially-focused techniques such as electrophysiological recordings. The following protocol describes a method of visualizing large-scale patterns of evoked hemodynamic activity in the cat auditory cortex using functional magnetic resonance imaging.

Imagem Funcional do Cortex Auditivo em gatos adultos usando de alta campo fMRI

Current knowledge of sensory processing in the mammalian auditory system is mainly derived from electrophysiological studies in a variety of animal ...

High Resolution Quantitative Synaptic Proteome Profiling of Mouse Brain Regions After Auditory Discrimination Learning

The molecular synaptic mechanisms underlying auditory learning and memory remain largely unknown. Here, the workflow of a proteomic study on auditory discrimination learning in mice is described. In this learning paradigm, mice are trained in a shuttle box Go/NoGo-task to discriminate between rising and falling frequency-modulated tones in order to avoid a mild electric foot-shock. The protocol involves the enrichment of synaptosomes from four brain areas, namely the auditory cortex, frontal cortex, hippocampus, and striatum, at different stages of training. Synaptic protein expression patterns obtained from trained mice are compared to na&#239;ve controls using a proteomic approach. To achieve sufficient analytical depth, samples are fractionated in three different ways prior to mass spectrometry, namely 1D SDS-PAGE/in-gel digestion, in-solution digestion and phospho-peptide enrichment. 
High-resolution proteomic analysis on a mass spectrometer and label-free quantification are used to examine synaptic protein profiles in phospho-peptide-depleted and phospho-peptide-enriched fractions of synaptosomal protein samples. A commercial software package is utilized to reveal proteins and phospho-peptides with significantly regulated relative synaptic abundance levels (trained/na&#239;ve controls). Common and differential regulation modes for the synaptic proteome in the investigated brain regions of mice after training were observed. Subsequently, meta-analyses utilizing several databases are employed to identify underlying cellular functions and biological pathways.

The identification of molecules and pathways controlling synaptic plasticity and memory is still a major challenge in neuroscience. Here, a workflow is described addressing the relative quantification of synaptic proteins supposedly involved in the molecular reorganization of synapses during learning and memory consolidation in an auditory learning paradigm.

Alta resolução quantitativa Synaptic Proteome Profiling de rato regiões cerebrais Depois Auditivo Discriminação Aprendizagem

The molecular synaptic mechanisms underlying auditory learning and memory remain largely unknown. Here, the workflow of a proteomic study on auditory ...

The Use of Trace Eyeblink Classical Conditioning to Assess Hippocampal Dysfunction in a Rat Model of Fetal Alcohol Spectrum Disorders

Neonatal rats were administered a relatively high concentration of ethyl alcohol (11.9% v/v) during postnatal days 4-9, a time when the fetal brain undergoes rapid organizational change and is similar to accelerated brain changes that occur during the third trimester in humans. This model of fetal alcohol spectrum disorders (FASDs) produces severe brain damage, mimicking the amount and pattern of binge-drinking that occurs in some pregnant alcoholic mothers. We describe the use of trace eyeblink classical conditioning (ECC), a higher-order variant of associative learning, to assess long-term hippocampal dysfunction that is typically seen in alcohol-exposed adult offspring. At 90 days of age, rodents were surgically prepared with recording and stimulating electrodes, which measured electromyographic (EMG) blink activity from the left eyelid muscle and delivered mild shock posterior to the left eye, respectively. After a 5 day recovery period, they underwent 6 sessions of trace ECC to determine associative learning differences between alcohol-exposed and control rats. Trace ECC is one of many possible ECC procedures that can be easily modified using the same equipment and software, so that different neural systems can be assessed. ECC procedures in general, can be used as diagnostic tools for detecting neural pathology in different brain systems and different conditions that insult the brain.

Trace eyeblink classical conditioning (ECC) was used to assess hippocampal-dependent associative learning in adult rats that were administered a high concentration (11.9% v/v) of alcohol during early neonatal brain development. In general, ECC procedures are sound diagnostic tools for detecting brain dysfunction across many psychological and biomedical settings.

O uso do condicionamento de rastreamento Eyeblink clássico para avaliar disfunção Hippocampal em um modelo do rato de transtornos do espectro do álcool Fetal

Neonatal rats were administered a relatively high concentration of ethyl alcohol (11.9% v/v) during postnatal days 4-9, a time when the fetal brain ...

Drosophila Courtship Conditioning As a Measure of Learning and Memory

Many insights into the molecular mechanisms underlying learning and memory have been elucidated through the use of simple behavioral assays in model organisms such as the fruit fly, Drosophila melanogaster. Drosophila is useful for understanding the basic neurobiology underlying cognitive deficits resulting from mutations in genes associated with human cognitive disorders, such as intellectual disability (ID) and autism. This work describes a methodology for testing learning and memory using a classic paradigm in Drosophila known as courtship conditioning. Male flies court females using a distinct pattern of easily recognizable behaviors. Premated females are not receptive to mating and will reject the male&#39;s copulation attempts. In response to this rejection, male flies reduce their courtship behavior. This learned reduction in courtship behavior is measured over time, serving as an indicator of learning and memory. The basic numerical output of this assay is the courtship index (CI), which is defined as the percentage of time that a male spends courting during a 10 min interval. The learning index (LI) is the relative reduction of CI in flies that have been exposed to a premated female compared to na&#239;ve flies with no previous social encounters. For the statistical comparison of LIs between genotypes, a randomization test with bootstrapping is used. To illustrate how the assay can be used to address the role of a gene relating to learning and memory, the pan-neuronal knockdown of Dihydroxyacetone phosphate acyltransferase (Dhap-at) was characterized here. The human ortholog of Dhap-at, glyceronephosphate O-acyltransferase (GNPT), is involved in rhizomelic chondrodysplasia punctata type 2, an autosomal-recessive syndrome characterized by severe ID. Using the courtship conditioning assay, it was determined that Dhap-at is required for long-term memory, but not for short-term memory. This result serves as a basis for further investigation of the underlying molecular mechanisms.

Drosophila Courtship Conditioning As a Measure of Learning and Memory

This protocol describes a Drosophila learning and memory assay called courtship conditioning. This classic assay is based on a reduction of male courtship behavior after sexual rejection by a non-receptive premated female. This natural form of behavioral plasticity can be used to test learning, short-term memory, and long-term memory.

Drosophila Courtship Acondicionamento Como Medida de Aprendizagem e Memória

Many insights into the molecular mechanisms underlying learning and memory have been elucidated through the use of simple behavioral assays in model ...

Quantifying Infra-slow Dynamics of Spectral Power and Heart Rate in Sleeping Mice

Three vigilance states dominate mammalian life: wakefulness, non-rapid eye movement (non-REM) sleep, and REM sleep. As more neural correlates of behavior are identified in freely moving animals, this three-fold subdivision becomes too simplistic. During wakefulness, ensembles of global and local cortical activities, together with peripheral parameters such as pupillary diameter and sympathovagal balance, define various degrees of arousal. It remains unclear the extent to which sleep also forms a continuum of brain states-within which the degree of resilience to sensory stimuli and arousability, and perhaps other sleep functions, vary gradually-and how peripheral physiological states co-vary. Research advancing the methods to monitor multiple parameters during sleep, as well as attributing to constellations of these functional attributes, is central to refining our understanding of sleep as a multifunctional process during which many beneficial effects must be executed. Identifying novel parameters characterizing sleep states will open opportunities for novel diagnostic avenues in sleep disorders.
We present a procedure to describe dynamic variations of mouse non-REM sleep states via the combined monitoring and analysis of electroencephalogram (EEG)/electrocorticogram (ECoG), electromyogram (EMG), and electrocardiogram (ECG) signals using standard polysomnographic recording techniques. Using this approach, we found that mouse non-REM sleep is organized into cycles of coordinated neural and cardiac oscillations that generate successive 25-s intervals of high and low fragility to external stimuli. Therefore, central and autonomic nervous systems are coordinated to form behaviorally distinct sleep states during consolidated non-REM sleep. We present surgical manipulations for polysomnographic (i.e., EEG/EMG combined with ECG) monitoring to track these cycles in the freely sleeping mouse, the analysis to quantify their dynamics, and the acoustic stimulation protocols to assess their role in the likelihood of waking up. Our approach has already been extended to human sleep and promises to unravel common organizing principles of non-REM sleep states in mammals.

Here, we present experimental and analytical procedures to describe the temporal dynamics of the neural and cardiac variables of non-REM sleep in mice, which modulate sleep responsiveness to acoustic stimuli.

Quantificando a dinâmica infra-lenta do poder espectral e da freqüência cardíaca em ratos dormindo

Three vigilance states dominate mammalian life: wakefulness, non-rapid eye movement (non-REM) sleep, and REM sleep. As more neural correlates of behavior ...

Desensitization and Recovery of Crayfish Photoreceptors Upon Delivery of a Light Stimulus

A method to study desensitization and recovery of crayfish photoreceptors is presented. We performed intracellular electrical recordings of photoreceptor cells in isolated eyestalks using the discontinuous single electrode-switched voltage-clamp configuration. First, with a razor blade we made an opening in the dorsal cornea to get access to the retina. Thereafter, we inserted a glass electrode through the opening, and penetrated a cell as reported by the recording of a negative potential. Membrane potential was clamped at the photoreceptor's resting potential and a light-pulse was applied to activate currents. Finally, the two light-flash protocol was employed to measure current desensitization and recovery. The first light-flash triggers, after a lag period, the transduction ionic current, which after reaching a peak amplitude decays towards a desensitized state; the second flash, applied at varying time intervals, assesses the state of the light-activated conductance. To characterize the light-elicited current, three parameters were measured: 1) latency (the time elapsed between light flash delivery and the moment in which current achieves 10% of its maximum value); 2) peak current; and 3) desensitization time constant (exponential time constant of the current decay phase). All parameters are affected by the first pulse.
To quantify recovery from desensitization, the ratio p2/p1 was employed versus time between pulses. p1 is the peak current evoked by the first light-pulse, and p2 is the peak current evoked by the second pulse. These data were fitted to a sum of exponential functions. Finally, these measurements were carried out as function of circadian time.

A protocol for the study of desensitization and sensitivity recovery of crayfish photoreceptors as a function of circadian time is presented.

Dessensibilização e recuperação de fotorreceptores de lagostins após a entrega de um estímulo leve

A method to study desensitization and recovery of crayfish photoreceptors is presented. We performed intracellular electrical recordings of photoreceptor ...

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and parenting. Importantly, mice can be trained to associate novel odors with specific behavioral responses to provide insight into olfactory circuit function. This protocol details the procedure for training mice on a Go/No-Go operant learning task. In this approach, mice are trained on hundreds of automated trials daily for 2&#8211;4 weeks and can then be tested on novel Go/No-Go odor pairs to assess olfactory discrimination, or be used for studies on how odor learning alters the structure or function of the olfactory circuit. Additionally, the mouse olfactory bulb (OB) features ongoing integration of adult-born neurons. Interestingly, olfactory learning increases both the survival and synaptic connections of these adult-born neurons. Therefore, this protocol can be combined with other biochemical, electrophysiological, and imaging techniques to study learning and activity-dependent factors that mediate neuronal survival and plasticity.

Here, we train mice on an associative learning task to test odor discrimination. This protocol also allows for studies on learning-induced structural changes in the brain.

Uma objetiva e reproduzível teste de aprendizagem olfactiva e discriminação em ratos

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and ...

Cochlear Implant Surgery and Electrically-evoked Auditory Brainstem Response Recordings in C57BL/6 Mice

Cochlear implants (CIs) are neuroprosthetic devices that can provide a sense of hearing to deaf people. However, a CI cannot restore all aspects of hearing. Improvement of the implant technology is needed if CI users are to perceive music and perform in more natural environments, such as hearing out a voice with competing talkers, reflections, and other sounds. Such improvement requires experimental animals to better understand the mechanisms of electric stimulation in the cochlea and its responses in the whole auditory system. The mouse is an increasingly attractive model due to the many genetic models available. However, the limited use of this species as a CI model is mainly due to the difficulty of implanting small electrode arrays. More details about the surgical procedure are therefore of great interest to expand the use of mice in CI research.
In this report, we describe in detail the protocol for acute deafening and cochlear implantation of an electrode array in the C57BL/6 mouse strain. We demonstrate the functional efficacy of this procedure with electrically-evoked auditory brainstem response (eABR) and show examples of facial nerve stimulation. Finally, we also discuss the importance of including a deafening procedure when using a normally hearing animal. This mouse model provides a powerful opportunity to study genetic and neurobiological mechanisms that would be of relevance for CI users.

Animal models of cochlear implants can advance knowledge of the technological bases of treating permanent sensorineural hearing loss with electrical stimulation. This study presents a surgical protocol for acute deafening and cochlear implantation of an electrode array in mice as well as the functional assessment with auditory brainstem response.

Cirurgia de implante coclear e gravações de resposta eletricamente evocado auditivo do tronco encefálico em camundongos C57BL/6

Cochlear implants (CIs) are neuroprosthetic devices that can provide a sense of hearing to deaf people. However, a CI cannot restore all aspects of ...