Nos gustaría dar las gracias a Helmut Scheu por la oportunidad de llevar a cabo nuestra investigación en el chiquero en el Hofgut Neumühle.

Todos los procedimientos fueron aprobados por el Comité de ética local (#23177-07/G10-1-010/G 15-15-011) y seguido de la Unión Europea y el Reglamento nacional alemán (europeo comunidades Consejo Directiva 86/609/ECC; Tierschutzgesetz).
Animales todos los procedimientos fueron realizados según el centro médico del Reglamento del Comité de cuidado de los animales de Johannes Gutenberg University Mainz.
1. configuración
<ol><li>Antes del experimento, verifique cualquier ruido en la línea y encontrar un lugar adecuado para la instalación y la antena. Ruido en la línea es visible como una onda de seno de 60 o 50 Hz. Nota: La colocación de la antena y sobre todo la distancia entre el transmisor y el receptor depende de la fuerza de transmisión del sistema. El sistema usado aquí es ajustable. Se ajustó a una potencia relativamente baja, con un rango de transmisión aproximadamente 3 m. Además, las vallas metálicas en el chiquero pueden atenuar la señal e interferencias. En este caso, es necesario colocar la antena dentro de la jaula del metal.</li>
	<li>Usar un tambor de cable para suministrar la instalación con energía de la línea. Conecte la computadora portátil, la unidad receptora y el convertidor de analógico a digital (si es necesario) para el sistema de telemetría específico que está siendo utilizado. Nota: El sistema de telemetría aquí envía datos digitales en el receptor. Esto puede ser diferente para otros sistemas.</li>
	<li>Coloque los electrodos, los adhesivos, los Q-tips y las toallitas, así como los bloques de mezcla, en una tabla separada.</li>
	<li>Preparar los electrodos con cables cortos. Para ello, los electrodos de corte y soldarlos de nuevo con una longitud lo más corta posible, dependiendo del tamaño del animal. Los cables deben ser lo suficientemente largos para conectar las posiciones de grabación deseado en la cabeza con la unidad de EEG telemetría, transmisión de los datos. Cables demasiado largos retrocedieron y cubiertos con elastómero de silicona adhesivo de la piel. Cables más largos que deben retrocedió hacen el parche de silicona más grande y más pesado.</li>
</ol>2. el cochinillo
<ol><li>Coger un cerdito agarrando la pierna o en el tórax. Mantenerlo y ser consciente de la defecación o micción.</li>
	<li>Si es necesario, marcar el cochinillo con un número.</li>
	<li>Envuelva el cochinillo en una toalla. El cochinillo se calmará. Ser conscientes de sobrecalentamiento el cochinillo.</li>
	<li>Sujete al lechón con una mano en el cuerpo o el antebrazo. Use la otra mano para sostener el hocico. Ser conscientes de sobrecalentamiento el cochinillo y asegúrese de que esté libre de respirar correctamente.</li>
</ol>3. electrodos
<ol><li>Tienen una segunda persona fije los electrodos.</li>
	<li>Limpiar la piel de la suciedad con agua o etanol. Si es necesario afeitarse la cabeza.</li>
	<li>Retire cualquier células muertas de la piel con un gel abrasivo de EEG y un q-Tip. Retirar el gel abrasivo después. Como alternativa, utilice papel de lija.</li>
	<li>Fije los electrodos autoadhesivos en la ubicación deseada. Coloque el electrodo de tierra por encima del cerebelo (entre las orejas) y el electrodo de referencia en la nariz. Coloque el electrodo de registro en la ubicación deseada. Nota: en este caso, un registro unipolar fue realizado, porque la referencia se colocó en una posición neutral (nariz). No se dispone de ningún sistema estandarizado para lechones hasta ahora. Aquí, una posición de grabación parietal fue utilizada (entre el ojo y el oído) en el hemisferio derecho del cerebro.</li>
	<li>Conecte los cables a la unidad de telemetría. Encienda la unidad. Según el sistema de telemetría, esto puede ser un interruptor magnético o una señal de frecuencia de radio despertador.</li>
	<li>Cubra la unidad de telemetría y todos los cables de electrodos, así como todo con goma de silicona adhesivo de dos componentes de la piel (véase Tabla de materiales). Al mezclar cantidades iguales de ambos componentes, el tiempo de curado será en el rango de 1 minuto de ojos y las pestañas no deben estar cubiertas por el caucho.</li>
	<li>Espere hasta que el caucho de silicón se cura completamente.</li>
	<li>Vuelva a colocar el lechón en la pocilga.</li>
	<li>Observar el cochinillo para ver si está mostrando signos de malestar durante un largo periodo de tiempo (varios minutos).</li>
</ol>4. la medida
<ol><li>Espere hasta que el cochinillo se ha recuperado y empieza a sincronizar su comportamiento con el de sus hermanos (alimentación, jugando, durmiendo), generalmente después de 30 s (figura 1).</li>
	<li>Espere a que las fases de sueño, si lo desea. El tiempo de grabación depende de la pregunta científica específica. Aquí, se utilizaron las sesiones de grabación de 10 minutos.</li>
	<li>Si la unidad de telemetría está cubierta por más de 2 otros lechones, la señal puede ser demasiado baja para el receptor. Empuje suavemente hacia los lechones, si duermen en la parte superior. Ser conscientes de la cerda; puede reaccionar agresivamente.</li>
	<li>Iniciar la grabación con el software de adquisición de datos (véase Tabla de materiales).</li>
</ol>5. final
<ol><li>Después de la grabación (generalmente varias horas), coger el cochinillo otra vez como se ha descrito en el paso 2. Ser conscientes de la cerda; puede reaccionar agresivamente.</li>
	<li>Suavemente, levante el caucho de silicón de un canto. Luego, retire el parche entero del caucho de silicón que contiene los electrodos y la unidad de telemetría. Tenga cuidado con los ojos del lechón.</li>
	<li>Vuelva a colocar el lechón en la pocilga.</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. 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Los autores no tienen nada que revelar.

Un paso crítico en el protocolo es el adecuado contacto con los electrodos, especialmente el electrodo de tierra, para lograr grabaciones estables con de poco ruido. Además, ya que los lechones son muy ágiles, es importante cubrir todo el sistema con caucho de silicona para proteger los electrodos y la unidad de telemetría. Además, si los experimentos se llevan a cabo en un establo con suelo de rejilla, ser cauteloso con los pequeños dispositivos o conectores.
En el caso de un agarre def...

Lechones son un modelo emergente para Neurociencia1. Con el fin de fortalecer la investigación traslacional, hemos inventado un método para registrar el EEG no invasivo, clínica de lechones libre2 (figura 1 y figura 2). Son los dos requisitos para una utilización traslacional de las grabaciones de EEG, con respecto a patrones de EEG asociados con la maduración cortical, una metodología no invasiva, comparable a la clínica y la abstinencia de sedantes o anestesia. El sistema de telemetría de un canal3 en combinación con electrodos autoadhesivos pueden fijarse en unos 5 minutos después, los lechones se recuperan rápidamente del procedimiento manejo y sincronizar su alimentación y para dormir el comportamiento de la otra lechones y la cerda.
Aunque ya hay intentos de utilizar las grabaciones de EEG no invasivo de animales sedados4, estudios de electroencefalograma (EEG) la mayoría de los animales se llevan a cabo con métodos invasivos. Estos métodos tienen efectos secundarios en relación con procesos inflamatorios alrededor de los electrodos implantados5,6 y en la mayoría de los casos, requiere una separación social de los animales debido a los componentes externos del sistema implantado de EEG. Por lo tanto, la traducción de estos datos en el contexto clínico es difícil. La necesidad de enfoques traslacionales se está convirtiendo en claro por el hecho de que todavía no se sabe cómo una maduración cerebral "normal" durante el desarrollo cortical temprano está representada por electroencefalografía clínica, invasiva7. Esta brecha de conocimiento es causada por problemas técnicos asociados con las grabaciones de EEG de recién nacidos prematuros bebés8. En sistemas modelo animales, patrones de desarrollo cortical temprana mejor son accesibles, ya que la mayoría de los animales nacen con un cerebro"prematuro" en comparación con el desarrollo cortical humano9. Además conservados patrones de desarrollo cortical a través de la especie2, recientemente se ha demostrado que las grabaciones de EEG de recién nacidos prematuros también pueden predecir el resultado clínico individual durante más de10,de vida11. El método descrito aquí es especialmente útil para los aspectos traslacionales de la neurociencia del desarrollo.

<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>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Laptop</td>
			<td style="width:309px;">Panasonic</td>
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			<td>Rugged laptop</td>
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		<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

El método permite la grabación de alta calidad los electroencefalogramas (EEGs) de libremente mover lechones directamente en el chiquero. Utilizamos un sistema de electroencefalografía telemétrica de un canal en combinación con electrodos estándar hidrogel auto-adhesivo. Los lechones se calmaron abajo sin el uso de sedantes. Después de su lanzamiento en la pocilga, los lechones se comportan normalmente, beber y dormir en el mismo ciclo como sus hermanos. Sus fases de sueño se utilizan para las grabaciones de EEG.

Pudimos grabar típicos patrones de EEG asociados con no sueño del REM, como ráfagas de huso o cepillos delta, mueva libremente lechones (figura 1 y figura 2). Principalmente estábamos interesados en patrones representativos durante no sueño del REM, pero las fases de sueño REM como12 con una amplitud muy baja también han sido registrados (figura 3). La fisiología y la ...

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

Noninvasive EEG Recordings from Freely Moving Piglets

Aquí, presentamos un protocolo de registro telemétricos electroencefalogramas (EEGs) de libremente mover lechones directamente en el chiquero sin el uso de un sedante, lo que es posible grabar patrones EEG típicos durante no sueño del REM, como huso estalla.

Grabaciones de EEG no invasivo se mueva libremente lechones

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 vistas.  Free University Berlin. Aquí, presentamos un protocolo de registro telemétricos electroencefalogramas (EEGs) de libremente mover lechones directamente en el chiquero sin el uso de un sedante, lo que es posible grabar patrones EEG típicos durante no sueño del REM, como huso estalla.

Video: 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.

Alta densidad de registros de EEG de los ratones se mueven libremente con poliamida basado microelectrodos

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

Investigación

Neurociencias

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.

Patch Clamp Las grabaciones de las células embrionarias de pez cebra 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.

Grabación Potenciales de Acción Individual Neuronas &quot;de que se mueven libremente Palomas en sus Tres Etapas del Aprendizaje

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 Ensayo para investigar las respuestas gustativas de individual, libre de movimiento abejorros (<em&gt; Bombus terrestris</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.

Grabación de EEG en Ratas Neonatales que se mueven libremente usando un método nuevo

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.

Grabaciones de potenciales evocadas visuales en ratones usando un multicanal no invasivo seco del cuero cabelludo 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 ...

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.

Grabaciones simultáneas de potenciales de campo Local Cortical, Electrocardiograma, electromiograma y respirar a ritmo de una rata libremente móvil

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.

Microdiálisis de aminoácidos excitatorios durante las grabaciones de EEG en mover libremente las ratas

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 largo plazo conflicto sensorial en comportarse libremente ratones

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 microdrive híbrido con sonda de optosilicio recuperable y tetroda para grabación de alta densidad de doble sitio en ratones en movimiento libre

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