Este trabajo fue apoyado por la Fundación de Ciencias Naturales de China (31171355) y la Fundación de Ciencias Naturales de Guangdong (S2011010003403, 2014A030313440).

El cuidado de los animales, el procedimiento quirúrgico y los procedimientos de registro estaban de acuerdo con las directrices para el Comité de Uso y Cuidado de Animales de la Universidad Normal de China del Sur. 1. Preparación del electrodo (Figura 1A-C) NOTA: Los loci de la clavija de la computadora son simplemente un contacto de la punta como parte de la interfaz de la señal en los dispositivos de la comunicación. Se compone de un conector macho que se conecta al conector hembra. <ol><li> Con cuidado, separe las clavijas macho y hembra de los loci de los clavijas de la computadora ( Figura 1A ) con la ayuda de una pinza. Conecte las clavijas macho y hembra entre sí para formar un electrodo y aplique cianoacrilato para crear una unión adhesiva fuerte ( Figura 1C ). </li><li> Coloque los electrodos en un vaso lleno de agua destilada y colóquelo en un limpiador ultrasónico durante 10 min. Muévalos a un horno de secado a 45 ° C durante 30 min. Esterilizar los electrodos con luz UV durante 30 min. </li></ol> 2. Procedimiento quirúrgico (Figura 1D-F) <ol><li> Preparar los instrumentos quirúrgicos esterilizados y los aparatos estereotáxicos. Anestesiar el cachorro de rata neonatal utilizando anestesia con isoflurano (2,5%) con aire. Cuando el cachorro está profundamente anestesiado, ajustar la dosis de isoflurano al 1,0%. Realizar una pinza de la cola o dedo del pie antes de la cirugía para asegurar la profundidad adecuada de la anestesia. </li><li> Fijar la cabeza del cachorro en el aparato estereotáxico colocando las barras de oído en los conductos auditivos y ligeramente apriete. NOTA: No apriete excesivamente las barras del oído, ya que el cráneo neonatal es muy suave. </li><li> Mantenga el campo quirúrgico estéril rociando todo el equipo con etanol al 70%. Haga una incisión de 15 mm en la cabeza usando un bisturí. Usando una pinza, tire suavemente del cuero cabelludo lejos de la línea media en las cuatro esquinas. Ponga un poco de algodón empapado en solución salina debajo de la piel para mantener la incisión abierta ( Figura 1D ). </li><li> Encuentra los puntos bregma y lambda en el cráneoY márquelos con un lápiz. Utilice una aguja de jeringa (26 G) para hacer dos agujeros de rebabas en la corteza prefrontal (PFC) y el hipocampo. NOTA: El PFC se localiza a +1.8 mm posterior a bregma y -0.5 mm lateral a la línea media, mientras que el hipocampo se encuentra a -2.0 mm anterior a bregma y ± 0.5 mm lateral a la línea media ( Figura 1D y E ). La profundidad del electrodo no debe estar a más de 2 mm por debajo de la superficie cortical para minimizar el daño cerebral. </li><li> Utilice una pinza para sujetar los electrodos e inserte los electrodos de referencia y de grabación en el PFC y el hipocampo, respectivamente. Aplique pomada de eritromicina alrededor del electrodo para evitar cualquier posible infección. Fijar el electrodo utilizando cianoacrilato. </li><li> Prepare el cemento dental acrílico para que tenga una consistencia pegajosa y viscosa. Aplique el cemento dental para cubrir los electrodos y el resto del cráneo. NOTA: Seque completamente el cráneo antes de aplicar el cemento dental. </li&gt; <li> Aplique ácido picrico al 5% sobre los electrodos para protegerlos. NOTA: Todo el procedimiento debe realizarse en una campana de bioseguridad para mantener condiciones estériles. </li><li> Retire el animal del marco estereotáxico e inyecte 300 μl de glucosa al 10% por vía subcutánea. Colóquelo sobre una manta caliente para su recuperación. Asegúrese de que el animal está caliente (37 ° C) y ambulatorio ( es decir, completamente recuperado). Administrar buprenorfina intraperitonealmente (0.05 mg / kg) para el dolor posquirúrgico. NOTA: No deje a un animal desatendido hasta que haya recuperado suficiente conciencia (es decir, comportamiento normal y movimiento). </li><li> Devuelve el cachorro a su jaula de casa con su madre después de que recupere la conciencia. Espere dos días hasta que el animal esté completamente recuperado. </li></ol> 3. Grabación EEG <ol><li> Después de la recuperación completa, conecte los electrodos implantados en el cráneo del cachorro al amplificador en su propia jaula. Conecte el amplificador a una toma analógica-digitalOnverter y conectar el convertidor a una computadora; Las líneas de conexión deben ser tratadas cuidadosamente para que no se enreden. </li><li> Seleccione una frecuencia de muestreo de al menos 10.000 Hz en la unidad de adquisición de datos para la grabación (el ancho de banda del transmisor es 1-100 Hz). Asegúrese de que los datos se muestreen correctamente. </li><li> Después de obtener la línea base de registro, inyectar el cabello intraperitonealmente con ácido cainico (KA) (2 mg / kg) para inducir ataques epilépticos. 15 min después de la inyección de KA observar y registrar las descargas epilépticas. Los productos de la incautación a través de KA suelen ser físicos. NOTA: La duración ictal-tónica es de aproximadamente 15,2 ± 0,9 s, la duración de la crisis es de aproximadamente 62 ± 5 s. Las convulsiones se pueden prevenir en la rata neonatal dando inyección de IP de hidrato de cloral (400 mg / kg). </li><li> Guarde los datos digitalizados y analícelos utilizando paquetes de software de procesamiento de señales, como spike2. Revelar el nivel de potencia de los diferentes componentes de frecuencia en la señal EEG neonatal por perfOrming un análisis de espectro de potencia. Calcule la potencia en un intervalo de tiempo de 1 minuto encontrando la amplitud de la raíz media cuadrada de 1 a 100 Hz (banda de EEG) 9 . </li></ol>

<ol>
	<li>Pereda, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+synapses+and+their+functional+interactions+with+chemical+synapses.">Electrical synapses and their functional interactions with chemical synapses.</a> Nat Rev Neurosci. 15 (4), 250-263 (2014).</li><li>Chorev, E., Epsztein, J., Houweling, A. R., Lee, A. K., Brecht, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+recordings+from+behaving+animals--going+beyond+spikes.">Electrophysiological recordings from behaving animals--going beyond spikes.</a> Curr Opin Neurobiol. 19 (5), 513-519 (2009).</li><li>Freeborn, D. L., McDaniel, K. L., Moser, V. C., Herr, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+electroencephalography+(EEG)+to+assess+CNS+changes+produced+by+pesticides+with+different+modes+of+action:+effects+of+permethrin,+deltamethrin,+fipronil,+imidacloprid,+carbaryl,+and+triadimefon.">Use of electroencephalography (EEG) to assess CNS changes produced by pesticides with different modes of action: effects of permethrin, deltamethrin, fipronil, imidacloprid, carbaryl, and triadimefon.</a> Toxicol Appl Pharmacol. 282 (2), 184-194 (2015).</li><li>Werhahn, K. J., Hartl, E., Hamann, K., Breimhorst, M., Noachtar, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Latency+of+interictal+epileptiform+discharges+in+long-term+EEG+recordings+in+epilepsy+patients.">Latency of interictal epileptiform discharges in long-term EEG recordings in epilepsy patients.</a> Seizure. 29, 20-25 (2015).</li><li>Staba, R. J., Stead, M., Worrell, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+biomarkers+of+epilepsy.">Electrophysiological biomarkers of epilepsy.</a> Neurotherapeutics. 11 (2), 334-346 (2014).</li><li>Nimmrich, V., Draguhn, A., Axmacher, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+Network+Oscillations+in+Neurodegenerative+Diseases.">Neuronal Network Oscillations in Neurodegenerative Diseases.</a> Neuromolecular Med. 17 (3), 270-284 (2015).</li><li>Zayachkivsky, A., Lehmkuhle, M. J., Dudek, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+Continuous+EEG+Monitoring+in+Small+Rodent+Models+of+Human+Disease+Using+the+Epoch+Wireless+Transmitter+System.">Long-term Continuous EEG Monitoring in Small Rodent Models of Human Disease Using the Epoch Wireless Transmitter System.</a> J Vis Exp. (101), e52554 (2015).</li><li>Zayachkivsky, A., Lehmkuhle, M. J., Fisher, J. H., Ekstrand, J. J., Dudek, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recording+EEG+in+immature+rats+with+a+novel+miniature+telemetry+system.">Recording EEG in immature rats with a novel miniature telemetry system.</a> J Neurophysiol. 109 (3), 900-911 (2013).</li><li>Dzhala, V. I., 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=NKCC1+transporter+facilitates+seizures+in+the+developing+brain.">NKCC1 transporter facilitates seizures in the developing brain.</a> Nat Med. 11 (11), 1205-1213 (2005).</li><li>Tucker, A. M., Aquilina, K., Chakkarapani, E., Hobbs, C. E., Thoresen, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+amplitude-integrated+electroencephalography+and+interburst+interval+in+the+rat.">Development of amplitude-integrated electroencephalography and interburst interval in the rat.</a> Pediatr Res. 65 (1), 62-66 (2009).</li><li>Savard, A., 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=Involvement+of+neuronal+IL-1beta+in+acquired+brain+lesions+in+a+rat+model+of+neonatal+encephalopathy.">Involvement of neuronal IL-1beta in acquired brain lesions in a rat model of neonatal encephalopathy.</a> J Neuroinflammation. 10, 110 (2013).</li><li>Cuaycong, M., 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=A+novel+approach+to+the+study+of+hypoxia-ischemia-induced+clinical+and+subclinical+seizures+in+the+neonatal+rat.">A novel approach to the study of hypoxia-ischemia-induced clinical and subclinical seizures in the neonatal rat.</a> Dev Neurosci. 33 (3-4), 241-250 (2011).</li></ol>

Los autores no declaran ningún conflicto de intereses.

Aquí, informamos de los procedimientos quirúrgicos y de registro para la adquisición de EEG en libre movimiento neonatal cachorros de rata por cable método ( Figura 3 ]. Se ha sugerido que el cachorro de rata P7-P12 está en la edad de desarrollo que corresponde a un recién nacido humano a término 10 , 11 . Es técnicamente difícil obtener datos de registro de EEG de alta calidad cuando se trabaja con cachorros de rata en este grupo de ed...

Está bien establecido que la comunicación continua entre las neuronas es necesaria para obtener la función cerebral normal. La comunicación interneuronal tiene lugar principalmente en las sinapsis, donde la información de una neurona se transmite a una segunda neurona. Esta transmisión sináptica está mediada por dos tipos de estructuras estructurales dedicadas: sinapsis eléctricas o químicas 1 . La electrofisiología es el campo que capta el potencial eléctrico producido durante la comunicación interneuronal que controla las funciones corporales y el comportamiento global 2 . EEG es el método más comúnmente utilizado entre muchas técnicas electrofisiológicas. EEG es una técnica utilizada para detectar cambios en las señales eléctricas producidas por estímulos internos o externos. Por otra parte, es una prueba esencial para el diagnóstico clínico y la predicción de los resultados de diversas afecciones neurológicas como epilepsia, Parkinson y enfermedad de AlzheimerE, así como los efectos de los agentes farmacológicos y toxicológicos 3 . Generalmente, un paciente epiléptico muestra hiperexcitabilidad y deterioro de la conectividad funcional dentro del cerebro; Estos se resumen como descargas epileptiformes interictal (IED) y pueden ser registrados por EEG en forma de picos agudos y transitorios; Olas agudas; Complejos de onda pico; O polyspikes 4 . La principal característica del cerebro epiléptico es la aparición espontánea de convulsiones epilépticas, que pueden registrarse tanto en el cuero cabelludo como en el parénquima cerebral para localizar el área cerebral responsable de las convulsiones 5 . Además, el EEG también tiene implicaciones muy importantes en trastornos neurodegenerativos como la enfermedad de Alzheimer (EA). La investigación sugiere que los registros alterados de EEG y redes oscilatorias deterioradas en pacientes con EA son comunes. Sin embargo, nuestro conocimiento sobre la patofisiología de las oscilaciones de la red en las enfermedades neurodegenerativas iEs sorprendentemente incompleta y necesita ser explorada más a fondo 6 . En este protocolo, hemos diseñado un electrodo simple con el que se puede grabar EEG para comprender la comunicación eléctrica en el cerebro normal y el patológico. La implantación quirúrgica en este método es más barato que otros procedimientos disponibles [ 7] . Además, este método puede utilizarse para registrar señales de EEG de alta calidad y estables para períodos de tiempo más largos ( es decir, 2-4 h todos los días durante 1 semana). Además, se utilizaron electrodos más ligeros (con un peso aproximado de 26 mg) que permiten a los animales comportarse de forma más natural 8 . Este método es ampliamente aplicable al estudio de EEG en cachorros de rata neonatal que requiere el amplificador y digitalizador, comúnmente utilizado en laboratorio de electrofisiología y no requiere ningún dispositivo adicional.

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			<td>Spike2</td>
			<td>Cambridge Electronic Design Ltd.</td>
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recording eeg in freely moving neonatal rats using a novel method

El EEG es un método útil para detectar la actividad eléctrica en el cerebro. Además, es una herramienta de diagnóstico ampliamente utilizada para diversas afecciones neurológicas, tales como epilepsia y trastornos neurodegenerativos. Sin embargo, técnicamente es difícil obtener registros de EEG en neonatos, ya que requiere un manejo especializado y un gran cuidado. Aquí, presentamos un nuevo método para registrar EEG en ratas neonatales (P8-P15). Diseñamos un electrodo simple y confiable usando localizadores de pin de computadora; Se puede implantar fácilmente en el cráneo de un cachorro de rata para grabar señales de EEG de alta calidad en el cerebro normal y epiléptico. Los cachorros recibieron una inyección intraperitoneal (ip) de la neurotoxina ácido cainico (KA) para inducir ataques epilépticos. La implantación quirúrgica realizada en este procedimiento es menos costosa que otros procedimientos EEG para neonatos. Este método permite registrar señales de EEG de alta calidad y estables durante más de una semana. Además, este procedimiento también se puede aplicar a adultos raSt y ratones para estudiar la epilepsia u otros trastornos neurológicos.

<html> Si los procedimientos quirúrgicos anteriores se llevan a cabo adecuadamente, se llevará a cabo con éxito un registro de EEG de un recién nacido de rata de rata de canal. 10 min después de la inyección de KA, un patrón regular de signos de comportamiento surgió en forma de movimientos irregulares y arañazos, temblores y pérdida de equilibrio. La Figura 2 muestra las trazas representativas del EEG bruto y las trazas expandidas ictal-tónicas e ictal-clónicas interictal. Re...

Watch this Scientific Journal Video about Recording EEG in Freely Moving Neonatal Rats Using a Novel Method at JoVE.com

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

Aquí, introducimos una técnica novedosa diseñada para registrar la electroencefalografía (EEG) en cachorros epilépticos neonatales de movimiento libre y describir sus procedimientos, características y aplicaciones. Este método permite registrar el EEG durante más de una semana.

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 ...

recording-eeg-in-freely-moving-neonatal-rats-using-a-novel-method

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

13.1K vistas.  South China Normal University.  Aquí, introducimos una técnica novedosa diseñada para registrar la electroencefalografía (EEG) en cachorros epilépticos neonatales de movimiento libre y describir sus procedimientos, características y aplicaciones. Este método permite registrar el EEG durante más de una semana. 

Video: Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

Novel Apparatus and Method for Drug Reinforcement

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction. Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in animal research to model various components of drug reinforcement, consumption, and addiction in humans. For this study, we used a novel approach to studying drug reinforcement in rats by combining traditional CPP and self-administration methodologies. We assembled an apparatus using two Med Associate operant chambers, sensory stimuli, and a Plexiglas-constructed neutral zone. These modifications allowed our experiments to encompass motivational aspects of drug intake through self-administration and drug-free assessment of drug/cue conditioning strength with the CPP test. In our experiments, rats self-administered cocaine (0.75 mg/kg/inj, i.v.) during either four (e.g., the "short-term") or eight (e.g., the "long-term") alternating-day sessions in an operant environment containing distinctive sensory cues (e.g., olfactory and visual). On the alternate days, in the other (differently-cued) operant environment, saline was available for self-infusion (0.1 ml, i.v.). Twenty-four hours after the last self-administration/cue-pairing session, a CPP test was conducted. Consistent with typical CPP findings, there was a significant preference for the chamber associated with cocaine self-administration. In addition, in animals undergoing the long-term experiment, a significant positive correlation between CPP magnitude and the number of cocaine-reinforced lever responses. In conclusion, this apparatus and approach is time and cost effective, can be used to examine a wide array of topics pertaining to drug abuse, and provides more flexibility in experimental design than CPP or self-administration methods alone.

Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in research to model various components of drug reinforcement, consumption, and addiction in humans. In this report, we combined traditional CPP and self-administration methods as a novel approach to studying drug reinforcement and addiction in rats.

Novela Aparato y método para el fortalecimiento de Drogas

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction.  Operant drug ...

Investigación

Neurociencias

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 ...

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

Un método general para la evaluación de incubación de sacarosa en ratas Deseo

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

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 ...

Extracellular Wire Tetrode Recording in Brain of Freely Walking Insects

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform natural behavior. We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allowed us to record activity from multiple neurons simultaneously while a tethered cockroach turned or altered walking speed. While a major advance, tethered preparations provide access to limited behaviors and often lack feedback processes that occur in freely moving animals. We now present a modified version of that technique that allows us to record from the central complex of freely moving cockroaches as they walk in an arena and deal with barriers by turning, climbing or tunneling. Coupled with high speed video and cluster cutting, we can now relate brain activity to various parameters of the movement of freely behaving insects.

We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allows us to monitor activity in individual units of tethered cockroaches. Here we present a modified version of that technique that allows us to also record brain activity in freely moving insects.

Extracelular alambre Tetrodo grabación en el cerebro de Libremente caminar Insectos

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform ...

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 ...

Noninvasive EEG Recordings from Freely Moving Piglets

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel telemetric electroencephalography system in combination with standard self-adhesive hydrogel electrodes. The piglets are calmed down without the use of sedatives. After their release into the pigpen, the piglets behave normally&#8212;they drink and sleep in the same cycle as their siblings. Their sleep phases are used for the EEG recordings.

Here, we present a protocol to record telemetric electroencephalograms (EEGs) from freely moving piglets directly in the pigpen without the use of a sedative, making it possible to record typical EEG patterns during non-REM sleep, like spindle bursts.

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 ...

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 ...

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, ...

Multi-Fiber Photometry to Record Neural Activity in Freely-Moving Animals

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and smaller functional subgroups, it becomes paramount to record from projections and/or genetically-defined subpopulations of neurons. Fiber photometry is an accessible and powerful approach that can overcome these challenges. By combining optical and genetic methodologies, neural activity can be measured in deep brain structures by expressing genetically-encoded calcium indicators, which translate neural activity into an optical signal that can be easily measured. The current protocol details the components of a multi-fiber photometry system, how to access deep brain structures to deliver and collect light, a method to account for motion artifacts, and how to process and analyze fluorescent signals. The protocol details experimental considerations when performing single and dual color imaging, from either single or multiple implanted optic fibers.

This protocol details how to implement and perform multi-fiber photometry recordings, how to correct for calcium-independent artifacts, and important considerations for dual-color photometry imaging.

Fotometría multifibra para registrar la actividad neuronal en animales en movimiento libre

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and ...

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

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