Gostaríamos de agradecer a Helmut Scheu para a oportunidade de realizar nossa pesquisa no chiqueiro em Hofgut Neumühle.

Todos os procedimentos foram aprovados pelo Comitê de ética local (#23177-07/G10-1-010/G 011-15-15) e seguiram a Europeia e a regulamentação nacional alemão (Europeu comunidades Conselho Directiva 86/609/CEE; Tierschutzgesetz).
Todos os procedimentos de animais foram efectuados de acordo com o centro médico de regulamentos da Comissão de cuidados com animais de Johannes Gutenberg-University Mainz.
1. instalação
<ol><li>Antes do experimento, verificar se há qualquer ruído de linha e encontrar um local adequado para a instalação e a antena. Ruído de linha é visível como uma onda de seno 50 ou 60 Hz. Nota: O posicionamento da antena e especialmente a distância entre o transmissor e o receptor depende da força de transmissão do sistema. O sistema usado aqui é ajustável. Ele foi ajustado para uma potência relativamente baixa, com uma faixa de transmissão aproximadamente 3 m. Além disso, as cercas metálicas no chiqueiro podem umedecer o sinal e causar interferências. Neste caso, é necessário colocar a antena dentro da jaula de metal.</li>
	<li>Use um tambor de cabo para fornecer a afinação com alimentação de linha. Conecte o laptop, a unidade receptora e o conversor analógico-digital (se necessário) para o sistema de telemetria específico que está sendo usado. Nota: O sistema de telemetria utilizado aqui enviado dados digitais para o receptor. Isto pode ser diferente para outros sistemas.</li>
	<li>Coloca os eletrodos, os adesivos, os cotonetes e lenços, bem como os blocos de misturando, em uma tabela separada.</li>
	<li>Prepare os eléctrodos com cabos curtos. Para fazer isso, os eletrodos de corte e solda-los novamente, com um comprimento mais curto possível, dependendo do tamanho do animal. Os cabos devem ser suficientemente compridas conectar as posições de gravação desejado na cabeça com a unidade de EEG telemetria, transmissão dos dados. Cabos muito longos devem ser recuou e cobertos com elastómero de silicone adesivo de pele. Cabos mais longos que devem ser recuou fazem o patch do silicone maiores e mais pesados.</li>
</ol>2. Leitão
<ol><li>Pega um leitão, agarrando-a na perna ou no tórax. Segurá-la e estar ciente de qualquer defecação ou micção.</li>
	<li>Se necessário, marca o leitão com um número.</li>
	<li>Enrole o leitão em uma toalha. O porquinho vai se acalmar. Esteja ciente de sobreaquecimento o leitão.</li>
	<li>Segure o leitão com uma mão no corpo ou no antebraço. Use a outra mão para segurar o focinho. Lembre-se de sobreaquecimento o leitão e certifique-se de que é livre para respirar corretamente.</li>
</ol>3. eletrodos
<ol><li>Tem uma segunda pessoa Coloque os eléctrodos.</li>
	<li>Limpe a pele de sujeira com água ou álcool etílico. Se necessário, raspe a cabeça.</li>
	<li>Remova as células mortas da pele com um gel abrasivo de EEG e um q-Tip. Remova o gel abrasivo depois. Como alternativa, use uma lixa.</li>
	<li>Fixar os eletrodos auto-adesivos no local desejado. Coloque o eléctrodo de terra acima do cerebelo (entre as orelhas) e o eletrodo de referência no nariz. Coloque o eléctrodo de gravação no local desejado. Nota: neste caso, uma gravação unipolar foi executada, porque a referência foi colocada em uma posição neutra (nariz). Não há nenhum sistema padronizado disponível para leitões até agora. Aqui, uma posição de gravação parietal foi usada (entre o olho e a orelha) no hemisfério direito do cérebro.</li>
	<li>Conecte os cabos da unidade de telemetria. Liga a unidade. Dependendo do sistema de telemetria usado, isso pode ser um interruptor magnético ou um sinal de despertar de rádio frequência.</li>
	<li>Cubra a unidade de telemetria e todos os cabos, bem como todos os eletrodos com a borracha de silicone adesivo de pele de dois componentes (ver Tabela de materiais). Através da mistura de quantidades iguais de ambos os componentes, o tempo de cura será no intervalo de 1 min. olhos e cílios não devem ser cobertos com a borracha.</li>
	<li>Espere até que a borracha de silicone está completamente curada.</li>
	<li>Coloque o leitão volta no chiqueiro.</li>
	<li>Observe o leitão para ver se ele está mostrando sinais de desconforto durante um longo período de tempo (vários minutos).</li>
</ol>4. medição
<ol><li>Espere até que o leitão se recuperou e começa a sincronizar o seu comportamento com o de seus irmãos (alimentando, brincando, dormindo), geralmente após 30 s (Figura 1).</li>
	<li>Espere por fases de sono, se desejado. O tempo de gravação varia de acordo com a questão científica específica. Aqui, utilizaram-se as sessões de gravação de 10 min.</li>
	<li>Se a unidade de telemetria é coberta por mais de 2 outros leitões, o sinal pode ser muito baixo para o receptor. Empurre suavemente os leitões embora se eles estão dormindo na parte superior. Lembre-se da proposta de serviço; Ele pode reagir agressivamente.</li>
	<li>Iniciar a gravação com o software de aquisição de dados (consulte a Tabela de materiais).</li>
</ol>5. revestimento
<ol><li>Depois de pegar a gravação (normalmente várias horas), o leitão novamente como foi descrito na etapa 2. Lembre-se da proposta de serviço; Ele pode reagir agressivamente.</li>
	<li>Levante suavemente a borracha de silicone em uma borda. Em seguida, remova o patch inteiro da borracha de silicone, que contém os eléctrodos e a unidade de telemetria. Cuidado com os olhos o leitão.</li>
	<li>Coloque o leitão volta no chiqueiro.</li>
</ol>

<ol>
	<li>Conrad, M. S., Sutton, B. P., Dilger, R. N., Johnson, R. W. <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+three-dimensional+magnetic+resonance+imaging-based+averaged+brain+collection+of+the+neonatal+piglet+(Sus+scrofa).">An in vivo three-dimensional magnetic resonance imaging-based averaged brain collection of the neonatal piglet (Sus scrofa).</a> PLoS ONE. 9 (9), e107650 (2014).</li><li>de Camp, N. V., Hense, F., Lecher, B., Scheu, H., Bergeler, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+for+preterm+cortical+development+using+non+invasive+clinical+EEG.">Models for preterm cortical development using non invasive clinical EEG.</a> Translational Neuroscience. 8, 211-224 (2017).</li><li>Lapray, D., Bergeler, J., Dupont, E., Thews, O., Luhmann, H. J., Barculo, D., Daniels, 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+novel+telemetric+system+for+recording+brain+activity+in+small+animals.">A novel telemetric system for recording brain activity in small animals.</a> Telemetry: Research, Technology and Applications. , 195-203 (2009).</li><li>Kim, D., Yeon, C., Kim, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+experimental+validation+of+a+dry+non-+invasive+multi-channel+mouse+scalp+EEG+sensor+through+visual+evoked+potential+recordings.">Development and experimental validation of a dry non- invasive multi-channel mouse scalp EEG sensor through visual evoked potential recordings.</a> Sensors. 17, 326 (2017).</li><li>Moshayedi, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationship+between+glial+cell+mechanosensitivity+and+foreign+body+reactions+in+the+central+nervous+system.">The relationship between glial cell mechanosensitivity and foreign body reactions in the central nervous system.</a> Biomaterials. 35, 3919-3925 (2014).</li><li>Barrese, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Failure+mode+analysis+of+silicon-based+intracortical+microelectrode+arrays+in+non-human+primates.">Failure mode analysis of silicon-based intracortical microelectrode arrays in non-human primates.</a> Journal of Neural Engineering. 10, 066014 (2013).</li><li>Hellstr&#246;m-Westas, L., Ros&#233;n, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electroencephalography+and+brain+damage+in+preterm+infants.">Electroencephalography and brain damage in preterm infants.</a> Early Human Development. 81, 255-261 (2005).</li><li>Lloyd, R. O., Goulding, R. M., Filan, P. M., Boylan, G. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Overcoming+the+practical+challenges+of+electroencephalography+for+very+preterm+infants+in+the+neonatal+intensive+care+unit.">Overcoming the practical challenges of electroencephalography for very preterm infants in the neonatal intensive care unit.</a> Acta Paediatrica. , 152-157 (2015).</li><li>Clancy, B., Finlay, B. L., Darlington, R. B., Anand, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extrapolating+brain+development+from+experimental+species+to+humans.">Extrapolating brain development from experimental species to humans.</a> Neurotoxicology. 28, 931-937 (2007).</li><li>Iyer, K. 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Os autores não têm nada para divulgar.

Um passo crítico no protocolo é o contato de pele adequados com os eléctrodos, especialmente o eléctrodo de terra, para conseguir gravações estáveis com baixo ruído. Além disso, desde que os leitões são muito ágeis, é importante cobrir todo o sistema com borracha de silicone para proteger os eléctrodos e a unidade de telemetria. Além disso, se as experiências são realizadas em um estábulo com um chão de ripas, seja cauteloso com pequenos dispositivos ou conectores.
No caso d...

Leitões são um sistema de modelo emergente de neurociência1. A fim de reforçar a investigação de translação, nós inventamos um método para gravar não-invasivo, clínicas EEGs de leitões desenfreada2 (Figura 1 e Figura 2). Duas condições prévias para uma utilização translacional de gravações de EEG, sobre padrões de EEG associados maturação cortical, são uma metodologia não-invasiva, comparável à clínica e a abstinência de sedativos ou anestesia. O sistema de telemetria de um canal3 em combinação com eletrodos auto-adesivos pode ser fixado em cerca de 5 min. depois, os leitões vão recuperar-se rapidamente do processo de manipulação e sincronizar sua alimentação e sono comportamento para que o outro leitões e a porca.
Embora já existam tenta usar gravações de EEG invasivo de animais sedado4, a maioria dos estudos de eletroencefalografia de animais são conduzidos com abordagens invasivas. Esses métodos têm efeitos colaterais sobre processos inflamatórios em torno de5,os eletrodos implantados6 e, na maioria dos casos, eles exigem uma separação social dos animais devido os componentes externos do sistema implantado de EEG. Portanto, a tradução destes dados ao contexto clínico é difícil. A necessidade de abordagens translacionais está se tornando claras pelo fato de que ainda não se sabe como uma maturação do cérebro "normais" durante o início do desenvolvimento cortical é representada por Eletroencefalografia clínica, não-invasivo7. Esta lacuna de conhecimento é causada pelos desafios técnicos associados com gravações de EEG de prematuros bebês8. Em sistemas modelo animal, padrões de desenvolvimento cortical precoce são mais acessíveis, desde que a maioria dos animais que nascem com um cérebro"prematuro" em comparação com o desenvolvimento cortical humano9. Além de conservadas padrões de desenvolvimento cortical em toda espécie2, recentemente mostrou que gravações de EEG de bebês prematuros também podem prever o resultado clínico individual durante posterior vida10,11. O método descrito aqui é especialmente útil para os aspectos translacionais de neurociência do desenvolvimento.

<table border="0" cellpadding="0" cellspacing="0" width="867">
	<tbody>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Disposable adhesive 
			surface silver/silver chloride electrodes</td>
			<td style="width:309px;">Spes 
			Medica S.r.l., Genova, Italy</td>
			<td style="width:119px;"></td>
			<td style="width:223px;">Self adhesive hydrogel electrode</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Abralyt HiCl</td>
			<td style="width:309px;">Easycap GmbH</td>
			<td style="width:119px;"></td>
			<td>Abrasive cream</td>
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			<td height="42" style="height:42px;width:216px;">Body Double fast</td>
			<td style="width:309px;">Smooth On Inc.</td>
			<td style="width:119px;"></td>
			<td>Skin adhesive silicone</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Telemetry system</td>
			<td style="width:309px;"></td>
			<td style="width:119px;"></td>
			<td>Internal development</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Picolog 1216</td>
			<td style="width:309px;">Pico Technology</td>
			<td style="width:119px;"></td>
			<td>AD converter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Laptop</td>
			<td style="width:309px;">Panasonic</td>
			<td style="width:119px;"></td>
			<td>Rugged laptop</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Receiver</td>
			<td style="width:309px;"></td>
			<td style="width:119px;"></td>
			<td>Internal development</td>
		</tr>
	</tbody>
</table>

noninvasive eeg recordings from freely moving piglets

O método permite a gravação de alta qualidade eletroencefalogramas (EEGs) um e outro se mova livremente leitões diretamente no chiqueiro. Nós usamos um sistema Telemétrico Eletroencefalografia um canal em combinação com eletrodos de hidrogel auto-adesivas padrão. Os leitões são calmos, sem o uso de sedativos. Após sua libertação no chiqueiro, os leitões se comportar normalmente — eles beber e dormir no mesmo ciclo como seus irmãos. Suas fases de sono são usados para as gravações de EEG.

Fomos capazes de gravar padrões de EEG típicos associados com não - sono REM, como rajadas de eixo ou escovas de delta, de mover-se livremente leitões (Figura 1 e Figura 2). Na maior parte estávamos interessados em padrões representativos durante não sono REM, mas as fases de sono REM, como12 , com uma amplitude muito baixa também têm sido registrados (Figura 3). A fi...

Watch this Scientific Journal Video about Noninvasive EEG Recordings from Freely Moving Piglets at JoVE.com

Noninvasive EEG Recordings from Freely Moving Piglets

Aqui, apresentamos um protocolo para registro Telemétrico eletroencefalogramas um e outro (EEGs) de movimentando-se livremente leitões diretamente no chiqueiro sem o uso de um sedativo, tornando possível gravar padrões típicos de EEG durante não sono REM, como eixo de rajadas.

Gravações de EEG não invasiva de leitões movimentando-se livremente

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel ...

noninvasive-eeg-recordings-from-freely-moving-piglets

Research

JoVE Journal

Neuroscience

7.2K visualizações.  Free University Berlin. Aqui, apresentamos um protocolo para registro Telemétrico eletroencefalogramas um e outro (EEGs) de movimentando-se livremente leitões diretamente no chiqueiro sem o uso de um sedativo, tornando possível gravar padrões típicos de EEG durante não sono REM, como eixo de rajadas.

Vídeo: Noninvasive EEG Recordings from Freely Moving Piglets

High-density EEG Recordings of the Freely Moving Mice using Polyimide-based Microelectrode

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a noninvasive brain monitoring tool in cognitive neuroscience as well as a diagnostic tool for epilepsy and sleep disorders in neurology. However, the underlying mechanism of EEG rhythm generation is still under the veil. Recently introduced polyimide-based microelectrode (PBM-array) for high resolution mouse EEG1 is one of the trials to answer the neurophysiological questions on EEG signals based on a rich genetic resource that the mouse model contains for the analysis of complex EEG generation process. This application of nanofabricated PBM-array to mouse skull is an efficient tool for collecting large-scale brain activity of transgenic mice and accommodates to identify the neural correlates to certain EEG rhythms in conjunction with behavior. However its ultra-thin thickness and bifurcated structure cause a trouble in handling and implantation of PBM-array. In the presented video, the preparation and surgery steps for the implantation of PBM-array on a mouse skull are described step by step. Handling and surgery tips to help researchers succeed in implantation are also provided.

In this article, we described the surgery procedure and handling tips for implantation of ultra-thin polyimide-based microelectrode array (PBM-array) on the mouse skull for acquisition of high-density encephalography (EEG) in a mouse model.

Gravações de alta densidade EEG dos Ratos movimentando-se livremente usando Polyimide baseado microeletrodos

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a ...

Neurociência

Patch Clamp Recordings from Embryonic Zebrafish Mauthner Cells

Mauthner cells (M-cells) are large reticulospinal neurons located in the hindbrain of teleost fish. They are key neurons involved in a characteristic behavior known as the C-start or escape response that occurs when the organism perceives a threat. The M-cell has been extensively studied in adult goldfish where it has been shown to receive a wide range of excitatory, inhibitory and neuromodulatory signals1. We have been examining M-cell activity in embryonic zebrafish in order to study aspects of synaptic development in a vertebrate preparation. In the late 1990s Ali and colleagues developed a preparation for patch clamp recording from M-cells in zebrafish embryos, in which the CNS was largely intact2,3,4. The objective at that time was to record synaptic activity from hindbrain neurons, spinal cord neurons and trunk skeletal muscle while maintaining functional synaptic connections within an intact brain-spinal cord preparation. This preparation is still used in our laboratory today. To examine the mechanisms underlying developmental synaptic plasticity, we record excitatory (AMPA and NMDA-mediated)5,6 and inhibitory (GABA and glycine) synaptic currents from developing M-cells. Importantly, this unique preparation allows us to return to the same cell (M-cell) from preparation to preparation to carefully examine synaptic plasticity and neuro-development in an embryonic organism. The benefits provided by this preparation include 1) intact, functional synaptic connections onto the M-cell, 2) relatively inexpensive preparations, 3) a large supply of readily available embryos 4) the ability to return to the same cell type (i.e. M-cell) in every preparation, so that synaptic development at the level of an individual cell can be examined from fish to fish, and 5) imaging of whole preparations due to the transparent nature of the embryos.

We have developed an intact brain-spinal cord preparation to record and monitor electrical activity via patch clamp recording from the Mauthner neurons and other reticulospinal cells in zebrafish embryos. Thus, we are able to record excitatory and inhibitory synaptic currents, voltage-gated channel activity and action potentials from key neurons in a developing embryo.

Grampo patch Gravações de Embryonic Cells Zebrafish Mauthner

Mauthner cells (M-cells) are large  reticulospinal neurons located in the hindbrain of teleost fish. They are key  neurons involved in a characteristic ...

Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed single-neuron activity while animals are acquiring an operantly conditioned response, or when that response is extinguished. But even in these cases, observation periods usually encompass only a single stage of learning, i.e. acquisition or extinction, but not both (exceptions include protocols employing reversal learning; see Bingman&#160;et al.1 for an example). However, acquisition and extinction entail different learning mechanisms and are therefore expected to be accompanied by different types and/or loci of neural plasticity.
Accordingly, we developed a behavioral paradigm which institutes three stages of learning in a single behavioral session and which is well suited for the simultaneous recording of single neurons&#39; action potentials. Animals are trained on a single-interval forced choice task which requires mapping each of two possible choice responses to the presentation of different novel visual stimuli (acquisition). After having reached a predefined performance criterion, one of the two choice responses is no longer reinforced (extinction). Following a certain decrement in performance level, correct responses are reinforced again (reacquisition). By using a new set of stimuli in every session, animals can undergo the acquisition-extinction-reacquisition process repeatedly. Because all three stages of learning occur in a single behavioral session, the paradigm is ideal for the simultaneous observation of the spiking output of multiple single neurons. We use pigeons as model systems, but the task can easily be adapted to any other species capable of conditioned discrimination learning.

Learning new stimulus-response associations engages a wide range of neural processes which are ultimately reflected in changing spike output of individual neurons. Here we describe a behavioral protocol allowing for the continuous registration of single-neuron activity while animals acquire, extinguish, and reacquire a conditioned response within a single experimental session.

Registro do potencial de ação simples Neurônios &#39;de livremente mover Pombos em três estágios da aprendizagem

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed ...

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees (Bombus terrestris)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from flowering plants. Only a few studies have described the gustatory responses of bees toward toxins in food, and these experiments have mainly used the proboscis extension response on restrained honey bees. Here, a new behavioral assay is presented for measuring the feeding responses of freely-moving, individual worker bumble bees to nutrients and toxins. This assay measures the amount of solution ingested by each bumble bee and identifies how tastants in food influence the microstructure of the feeding behavior.
The solutions are presented in a microcapillary tube to individual bumble bees that have been previously starved for 2-4 hr. The behavior is captured on digital video. The fine structure of the feeding behavior is analyzed by continuously scoring the position of the proboscis (mouthparts) from video recordings using event logging software. The position of the proboscis is defined by three different behavioral categories: (1) proboscis is extended and in contact with the solution, (2) proboscis is extended but not in contact with the solution and (3) proboscis is stowed under the head. Furthermore the speed of the proboscis retracting away from the solution is also estimated.
In the present assay the volume of solution consumed, the number of feeding bouts, the duration of the feeding bouts and the speed of the proboscis retraction after the first contact is used to evaluate the phagostimulatory or the deterrent activity of the compounds tested.
This new taste assay will allow researchers to measure how compounds found in nectar influence the feeding behavior of bees and will also be useful to pollination biologists, toxicologists and neuroethologists studying the bumble bee&#39;s taste system.

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees Bombus terrestris

A novel behavioral assay is described for investigating the short term gustatory responses of the mouthparts of freely-moving bumble bees (Bombus terrestris) toward nutrients and toxins in solution.

A Novel Behavioral Ensaio investigar as gustativas de um indivíduo, livre-se deslocam Bumble abelhas (<em&gt; Terrestris do Bombus</em&gt;)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from ...

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such as epilepsy and neurodegenerative disorders. However, it is technically difficult to obtain EEG recordings in neonates as it requires specialized handling and great care. Here, we present a novel method to record EEG in neonatal rat pups (P8-P15). We designed a simple and reliable electrode using computer pin loci; it can be easily implanted into the skull of a rat pup to record high-quality EEG signals in the normal and epileptic brain. Pups were given an intraperitoneal (i.p.) injection of the neurotoxin kainic acid (KA) to induce epileptic seizures. The surgical implantation performed in this procedure is less expensive than other EEG procedures for neonates. This method allows one to record high-quality and stable EEG signals for more than 1 week. Furthermore, this procedure can also be applied to adult rats and mice to study epilepsy or other neurological disorders.

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

Gravação de EEG em Ratos Neonatais de Movimentação Livre Usando um Novo Método

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

Visual Evoked Potential Recordings in Mice Using a Dry Non-invasive Multi-channel Scalp EEG Sensor

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable because of the plunger-spring-barrel structural facet and mechanical strengths resulting from metal materials. The whole process for acquiring the VEP responses in vivo from a mouse consists of four steps: (1) sensor assembly, (2) animal preparation, (3) VEP measurement, and (4) signal processing. This paper presents representative measurements of VEP responses from multiple mice with a submicro-voltage signal resolution and sub-hundred millisecond temporal resolution. Although the proposed method is safer and more convenient compared to other previously reported animal EEG acquiring methods, there are remaining issues including how to enhance the signal-to-noise ratio and how to apply this technique with freely moving animals. The proposed method utilizes easily available resources and shows a repetitive VEP response with a satisfactory signal quality. Therefore, this method could be utilized for longitudinal experimental studies and reliable translational research exploiting non-invasive paradigms.

We designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable. This paper describes the whole process from manufacturing the proposed EEG electrode to signal processing of visual evoked potential (VEP) signals measured on a mouse scalp using a dry non-invasive multi-channel EEG sensor.

Gravações de potenciais evocadas visuais em ratos usando um não-invasivo seco multi-canal do couro cabeludo Sensor de EEG

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

Gravações simultâneas de potenciais de campo corticais Local, eletrocardiograma, eletromiograma e respirar o ritmo de um rato andando livremente

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain interstitial space with the behavior and/or with the specific outcome of a pathology (e.g., seizures for epilepsy). When studying epilepsy, the microdialysis technique is often combined with short-term or even long-term video-electroencephalography (EEG) monitoring to assess spontaneous seizure frequency, severity, progression and clustering. The combined microdialysis-EEG is based on the use of several methods and instruments. Here, we performed in vivo microdialysis and continuous video-EEG recording to monitor glutamate and aspartate outflow over time, in different phases of the natural history of epilepsy in a rat model. This combined approach allows the pairing of changes in the neurotransmitter release with specific stages of the disease development and progression. The amino acid concentration in the dialysate was determined by liquid chromatography. Here, we describe the methods and outline the principal precautionary measures one should take during in vivo microdialysis-EEG, with particular attention to the stereotaxic surgery, basal and high potassium stimulation during microdialysis, depth electrode EEG recording and high-performance liquid chromatography analysis of aspartate and glutamate in the dialysate. This approach may be adapted to test a variety of drug or disease induced changes of the physiological concentrations of aspartate and glutamate in the brain. Depending on the availability of an appropriate analytical assay, it may be further used to test different soluble molecules when employing EEG recording at the same time.

Here, we describe a method for in vivo microdialysis to analyze aspartate and glutamate release in the ventral hippocampus of epileptic and non-epileptic rats, in combination with EEG recordings. Extracellular concentrations of aspartate and glutamate may be correlated with the different phases of the disease.

Microdialysis de aminoácidos excitatórios durante as gravações de EEG em ratos movimentando-se livremente

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain ...

Long-term Sensory Conflict in Freely Behaving Mice

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning in mice. By permanently wearing a device fixed on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages. Therefore, this protocol readily enables the study of the visual system and multisensory interactions over an extended timeframe that would not be accessible otherwise. In addition to lowering the experimental costs of long-term sensory learning in naturally behaving mice, this approach accommodates the combination of in vivo and in vitro experiments. In the reported example, video-oculography is performed to quantify the vestibulo-ocular reflex (VOR) and optokinetic reflex (OKR) before and after learning. Mice exposed to this long-term sensory conflict between visual and vestibular inputs presented a strong VOR gain decrease but exhibited few OKR changes. Detailed steps of device assembly, animal care, and reflex measurements are hereby reported.

The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning. By permanently wearing a fixed device on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages.

A longo prazo conflito sensorial em livremente, comportando-se ratos

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for ...

Hybrid Microdrive System with Recoverable Opto-Silicon Probe and Tetrode for Dual-Site High Density Recording in Freely Moving Mice

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, conventional microdrive designs often only allow use of one type of electrode to record from single or multiple regions, limiting the yield of single-unit or depth profile recordings. It also often limits the ability to combine electrode recordings with optogenetic tools to target pathway and/or cell type specific activity. Presented here is a hybrid microdrive array for freely moving mice to optimize yield and a description of its fabrication and reuse of the microdrive array. The current design employs nine tetrodes and one opto-silicon probe implanted in two different brain areas simultaneously in freely moving mice. The tetrodes and the opto-silicon probe are independently adjustable along the dorsoventral axis in the brain to maximize the yield of unit and oscillatory activities. This microdrive array also incorporates a set-up for light, mediating optogenetic manipulation to investigate the regional- or cell type-specific responses and functions of long-range neural circuits. In addition, the opto-silicon probe can be safely recovered and reused after each experiment. Because the microdrive array consists of 3D-printed parts, the design of microdrives can be easily modified to accommodate various settings. First described is the design of the microdrive array and how to attach the optical fiber to a silicon probe for optogenetics experiments, followed by fabrication of the tetrode bundle and implantation of the array into a mouse brain. The recording of local field potentials and unit spiking combined with optogenetic stimulation also demonstrate feasibility of the microdrive array system in freely moving mice.

This protocol describes the construction of a hybrid microdrive array that allows implantation of nine independently adjustable tetrodes and one adjustable opto-silicon probe in two brain regions in freely moving mice. Also demonstrated is a method for safely recovering and reusing the opto-silicon probe for multiple purposes.

Sistema híbrido do Microdrive com a ponta de prova recuperável do opto-silicone e tetrode para a gravação high-density do duplo-local em ratos livremente moventes

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, ...

Gravações de EEG não invasiva de leitões movimentando-se livremente

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