MSB is supported by the Alcohol Beverage Medical Research Foundation, a Center for Translational Research Award (UL1 TR000058), the National Institutes for Alcohol Abuse and Alcoholism (P50 AA022537), and startup funds provided by the Virginia Higher Education Equipment Trust Fund and the VCU School of Medicine.

<ol>
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None of the authors have competing or conflicting interests.

El procedimiento que aquí se presenta es un medio eficaz para la fabricación de cánulas microinyección y microinyectores que ayudarán en la aclaración de los sustratos moleculares de la conducta motivada. Este método ofrece varias ventajas. En primer lugar, mediante la fabricación de los propios implantes y microinyectores, nuevos parámetros experimentales se pueden optimizar con rapidez, es decir, uno no necesita esperar a que los componentes a medida que lleguen. En segundo lugar, debido al pequeño diámetro de la cánula, más cánulas pueden ser implantados simultáneamente. Esto acorta el tiempo quirúrgico requerido, lo que puede mejorar la supervivencia, y también permite que múltiples implantes por animal. En tercer lugar, el software utilizado para controlar las cámaras operantes acomoda fácilmente Escala de tasa progresivas desde un paradigma proporción fija se puede convertir rápidamente a un paradigma relación progresiva mediante la simple aplicación de una lista de parámetros de transición caso de que contiene el programa de refuerzo deseado. Serampliamente útil, un procedimiento de microinyección genérico se presentó que debería ser ampliamente aplicable para la microinyección de casi cualquier reactivo disponible actualmente. En consecuencia, esperamos que esta técnica seguirá siendo de gran utilidad similar en el futuro con pequeñas modificaciones. Por cambiando sólo unas pocas variables, este enfoque se puede aplicar a un amplio número de reactivos. Los parámetros que se pueden manipular más comúnmente incluyen la longitud que el microinyector sobresale de la cánula, el volumen de la inyección, y la velocidad de inyección. Por ejemplo, uno puede desear el inyector sobresalga más lejos de la punta de la cánula para evitar la cicatriz glial que normalmente se forma alrededor de los implantes crónicas. Adicionalmente, se puede desear para inyectar un volumen mayor. Para microinyecciones virus del estriado, un volumen de 1 l se utiliza normalmente y este volumen se inyecta típicamente durante un período de tiempo más largo (con frecuencia 7 - 10 min plus 3 - 10 min tiempo de difusión adicional) en comparación con que el usod para los reactivos farmacológicos (típicamente 0,3 - 0,5 l más de 2 - 3 minutos más 1 - 3 min tiempo de difusión adicional). El usuario debe consultar la literatura y / o empíricamente determinar los parámetros más adecuados para sus necesidades. Independientemente, el éxito de este procedimiento depende críticamente de 4 variables: 1) la longitud de la cánula, 2) longitud microinyector, 3) calidad de patrón de pulverización microinyector, y 4) la integridad del sistema antes de la inyección. Debido a la ubicación microinyección depende de la profundidad que el microinyector sobresale de la cánula, es imperativo que tanto cánulas (Paso 1.2.8) y la longitud microinyector (post-flexión, Paso 2.2.1) son ambos precisamente conocida y uniforme entre todos los sujetos . Esto puede ser fácilmente controlado por fácilmente rechazar cualquier utensilio que no es la longitud requerida en la final de la re-medición. Por otra parte, la ubicación de inyección sólo puede predecirse si se produce inmediatamente por debajo de la cánula guía. Por lo tanto, cualquier microinyector que el DOEs No rocíe un largo arroyo, bien en las pruebas (Pasos 2.4.6) debe ser rechazada. Una inyección de calidad también está relacionada con la integridad del sistema antes de la inyección. Si después de dispensar toda el agua del inyector (antes de llenar con el reactivo) se observan varios puntos en el laboratorio de borrado, entonces una fuga necesita ser remediado (Nota sobre el paso 2.4.8). Además, si la burbuja (Paso 2.4.9) que separa el medicamento del agua en la tubería PE20 es no uno, burbuja única (después de llenar el microinyector con el reactivo), entonces el inyector está parcialmente obstruido. Esta obstrucción puede prevenir o desviar la inyección. Esto también se puede remediar fácilmente (Nota en el paso 2.4.8). Si uno desea microinyectar mientras el animal está en el marco estereotáxico hay tres alternativas. En primer lugar, se podría aumentar la longitud del collar microinyector tal que puede ser sostenida firmemente por el manipulador estereotáxico y también se extiende lo suficiente para permitir la conexión a un tubo de PE20. En segundo lugar, ene temporalmente podría implantar una cánula y utilizar el microinyector norma que aquí se presenta. En tercer lugar, se podría utilizar dibujado y pipetas de vidrio pulido. 16,17 Una limitación importante del procedimiento que aquí se presenta es que se realiza mejor en ratas bien manejados, que están familiarizados con el procedimiento. Las ratas utilizadas para los datos que se describen en la sección de resultados requieren procedimientos especiales de manipulación debido a que el mismo investigador maneja las ratas sobre una base diaria durante más de 2 meses. Esto incluyó la observación diaria y la manipulación del implante quirúrgico durante al menos 2 semanas. Sin embargo, las ratas se habituaron rápidamente pueden por una serie de técnicas que se utilizan para antes del ensayo de inhibición pre-pulso, que puede verse afectada por el estrés. Estas técnicas especiales de habituación han sido muy bien detallado anteriormente. 43 Además de estos procedimientos, es aconsejable que las ratas se habituaron al procedimiento de microinyección donde se utilizan microinyectores acortados duinyecciones anillo &#39;farsa&#39;. Durante estas inyecciones simuladas, es fundamental que el microinyector no sobresale en el tejido con el fin de limitar el daño a los tejidos. En otras palabras, el microinyector debe ser de no más de 14 mm se inclinó. Por lo tanto, la habituación a fondo requerida para la aplicación óptima de esta técnica podría ser visto como una limitación. Si bien existen varios paradigmas de comportamiento para medir la motivación, la relación progresiva se utiliza comúnmente para cuantificar el esfuerzo que el sujeto está dispuesto para ejercer para obtener un reforzador. El paradigma relación progresiva produce una medida conocida como punto de interrupción, que a menudo se define como el número máximo de pulsaciones de palanca en la última relación completado;.. Es decir, máxima respuesta que genera un reforzador 21 La relación progresiva es sensible a reforzador magnitud. Por ejemplo, el aumento de las dosis de cocaína (o sacarosa) producen un mayor punto de interrupción y la cocaína inferior (o sacarosa) dosis producen una breakp menormixto. 21,22 En consecuencia, punto de interrupción es un proxy utilizado habitualmente para la motivación y / o eficacia de refuerzo. 21,23-26 Porque la intención del punto de interrupción es determinar cuando el animal deja de responder, un parámetro importante del paradigma de relación progresiva es duración de la sesión. Duración de la sesión finitos puede poner una tapa falsa en los valores de punto de interrupción y esto puede ser exacerbada por los pre-tratamientos que disminuyen de forma anormal la tasa de auto-administración o que pausa aumento post-refuerzo. Este factor de confusión puede ser superada por cualquier número de enfoques;.. Por ejemplo, las sesiones que terminan cuando el animal ha retenido responder por algún múltiplo del intervalo promedio entre infusión 44 Una variante más comúnmente aplicada de este enfoque es a terminar sesiones vez de responder tiene sido retenida por algún valor determinado empíricamente que se mantiene constante a través de temas. Hemos proporcionado el método para aplicar este enfoque en el Paso 4.4.9.11.

Rodents and humans respond in remarkably similar ways to behaviorally relevant stimuli1-3. This suggests that rodents are appropriate subjects for elucidating the molecular substrates of behavior and complex psychiatric conditions4. Understanding the molecular substrates of complex behavioral processes, such as motivation, frequently requires brain microinjection. Both the brain microinjection technique and a primary motivation assay will be presented here. Rats will be used as subjects, but these procedures can readily be adapted to well-handled mice. Included herein are procedures for the manufacture of the required cannulae, obturators (dummy cannulae or stylets), and microinjectors. The method presented is significantly more flexible and more cost-efficient than prefabricated implements. This flexibility will prove valuable when optimizing conditions. Importantly, because the microinjection procedure can be used to test a myriad of hypotheses; the techniques presented here should be broadly applicable. For example, receptor ligands can be microinjected to understand neurochemistry3,5,6; cell-permeable peptides and small-molecules can be microinjected to understand intracellular signaling pathways7-10; toxins, ion channel blockers, or antagonist cocktails can be microinjected to understand circuitry1,11,12.
While the generic protocol presented here can be readily adapted by the user for their particular needs, the procedure is particularly well suited for behavioral assays since microinjection occurs in awake rodents that are only under mild hand restraint. No anesthesia or special restraints are required. This is possible because the brain itself lacks pain sensation. However, if anesthesia is not used, microinjection must occur through cannulae that were previously stereotaxically implanted. This is because nociceptors are present on the scalp, meninges,13 which are the membranes surrounding the brain, and the periosteum,14 which is the membrane covering the skull. It should be noted that microinjection under anesthesia is sometimes desirable. One example is when the virus is being injected, and one may wish to inject virus directly through either stainless steal needles15 or glass pipettes because this can reduce tissue damage and improve transduction efficiency.16,17 The microinjectors described below can be modified for this purpose and suggestions on how to do this can be found in the Discussion. Because other JoVE articles have demonstrated stereotaxic brain cannula implantation,18-20 these procedures will not be covered here.
We present these microinjection procedures together with an assay that quantifies motivation. Several rodent models of motivated behavior are currently in use, such as the runway box and barrier scaling. Here, we describe how to use an operant progressive ratio schedule of reinforcement to quantify motivation where operant responding is being maintained by a reinforcer. Responding on the progressive ratio is responsive to reinforcer magnitude.21,22 Accordingly, this assay is routinely used as a proxy for motivation and/or reinforcing efficacy. 21,23-30 Because several excellent reviews have covered this topic in detail,21,24 we will focus mainly on practical concerns.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Cannula Tubing</td>
			<td>Amazon Supply/ Small Parts</td>
			<td>HTXX-26T-60</td>
			<td>26 gauge, Hypotube S/S 316-TW 26GA</td>
		</tr>
		<tr>
			<td>Obturator</td>
			<td>Amazon Supply/ Small Parts</td>
			<td>GWXX-0080-30-05</td>
			<td>33 gauge, Wire S/S 316LVM 0.008 IN</td>
		</tr>
		<tr>
			<td>Microinjector Wire</td>
			<td>MicroGroup</td>
			<td>33RW 304</td>
			<td>33 gauge</td>
		</tr>
		<tr>
			<td>Super Glue</td>
			<td>Loctite</td>
			<td>3924AC</td>
			<td>Liquid, Non-gel, can be autoclaved</td>
		</tr>
		<tr>
			<td>Microinjector Plastic Tubing</td>
			<td>Becton Dickson</td>
			<td>427406</td>
			<td>PE20</td>
		</tr>
		<tr>
			<td>Medium Weight Hemostats</td>
			<td>World Precision Instruments</td>
			<td>501241-G</td>
			<td></td>
		</tr>
		<tr>
			<td>Ruler</td>
			<td>Fisher</td>
			<td>09-016</td>
			<td>150 mm</td>
		</tr>
		<tr>
			<td>#7 Forceps</td>
			<td>Stoelting</td>
			<td>52100-77</td>
			<td>Dumont, Dumostar</td>
		</tr>
		<tr>
			<td>Rotary Tool</td>
			<td>Dremmel</td>
			<td>285</td>
			<td>Two-speeds</td>
		</tr>
		<tr>
			<td>Cut-off Disc</td>
			<td>McMaster Carr</td>
			<td>3602</td>
			<td>15/16&#34; x 0.025&#34;</td>
		</tr>
		<tr>
			<td>Microinjection Pump</td>
			<td>Harvard Apparatus</td>
			<td>PhD 2000</td>
			<td></td>
		</tr>
		<tr>
			<td>1 ul Glass Syringe</td>
			<td>Hamilton</td>
			<td>7001KH</td>
			<td>Needle Style: 25s/2.75&#34;/3</td>
		</tr>
		<tr>
			<td>Cotton Tipped Applicator</td>
			<td>Fisher</td>
			<td>23-400-101</td>
			<td></td>
		</tr>
		<tr>
			<td>Lab Wipes</td>
			<td>Kimwipes</td>
			<td>34133</td>
			<td></td>
		</tr>
		<tr>
			<td>Operant Software</td>
			<td>Coulbourn</td>
			<td>Graphic State</td>
			<td></td>
		</tr>
		<tr>
			<td>Operant Chambers</td>
			<td>Coulborun</td>
			<td>Habitest</td>
			<td></td>
		</tr>
	</tbody>
</table>

rodent brain microinjection to study molecular substrates of motivated behavior

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as appropriate models, partly because the response to behaviorally relevant stimuli and the circuitry parsing stimulus-action outcomes is astonishingly similar between humans and rodents. In studying molecular substrates of complex behaviors, the microinjection of reagents that modify, augment, or silence specific systems is an invaluable technique. However, it is crucial that the microinjection site is precisely targeted in order to aid interpretation of the results. We present a method for the manufacture of surgical implements and microinjection needles that enables accurate microinjection and unlimited customizability with minimal cost. Importantly, this technique can be successfully completed in awake rodents if conducted in conjunction with other JoVE articles that covered requisite surgical procedures. Additionally, there are many behavioral paradigms that are well suited for measuring motivation. The progressive ratio is a commonly used method that quantifies the efficacy of a reinforcer to maintain responding despite an (often exponentially) increasing work requirement. This assay is sensitive to reinforcer magnitude and pharmacological manipulations, which allows reinforcing efficacy and/ or motivation to be determined. We also present a straightforward approach to program operant software to accommodate a progressive ratio reinforcement schedule.

Aquí se ilustran ejemplos de los resultados que se pueden obtener con el procedimiento anterior. Primero que se muestra es una zona típica que se transfectadas por los vectores virales (Figura 3). En general, se obtiene un volumen de ~ 1 transfectadas mm 3 en el tejido estriatal. El volumen de la región transfectadas se puede cuantificar a través de secciones en serie, utilizando el método de Cavalieri 32 Es importante destacar que, el volumen transfectadas depende de muchos factores tales como el tipo de tejido que se está transfectada.; serotipo viral; promotor; velocidad y el volumen inyectado; número de partículas inyectadas; pH inyectado; y si hiperosmolares reactivos, tales como manitol, se utilizaron 33,34. Típicamente, nos microinyectar 10 12 partículas / ml, 1 l / min secundarios sobre 10, y permiten 7 min adicionales antes de reemplazar obturadores. Además, el inyectado es generalmente alrededor de pH 8. A continuación, se muestra que la motivación puede ser manipulado por cualquiera de la expresión del transgen (Figura 4)o microinyección reactivo farmacológico (Figura 5). En la Figura 4, expresamos un receptor Diseñador Exclusivamente activado por las drogas de diseño (DREADDs). La región codificante del receptor de DREADD fue seguido por un sitio de entrada de ribosoma interno, y una casete mCitrine. El mCitrine permite la visualización conveniente de células transfectadas. El DREADD se acopló a la proteína G heterotrimeric Gαq. La activación de la DREADD acoplado Gαq puede estimular astrocitos, 28,35 y el propio DREADD puede ser activado por la administración sistémica de clozapina-N-óxido (CNO, 3 mg / kg, ip). 14,22,36 Las ratas fueron entrenadas para palanca de responder para el refuerzo de etanol, donde 3 prensas de palanca produjeron 1 oportunidad de beber durante las diarias sesiones de 1 h más de 60 días consecutivos. A continuación, las ratas fueron obligados a la abstinencia, y DREADDs acoplados Gαq se expresaron en el núcleo accumbens astrocitos centrales. Después de 3 semanas de abstinencia, la motivación para la auto-admi<html> etanol Nister se midió por punto de interrupción. 21,27-30 La activación del núcleo accumbens astrocitos fundamentales, a través de la administración sistémica CNO, disminución de la motivación de las ratas para reanudar etanol autoadministración después de la abstinencia en comparación con el vehículo. Es importante destacar que, CNO no tuvo ningún efecto en una cohorte igualmente capacitado que expresaba la proteína fluorescente verde en lugar de la DREADD. <img alt="Figura 3" src="/files/ftp_upload/53018/53018fig3.jpg" /> Figura 3. región estriatal ventral Representante transfectadas por el virus de microinyectados. Esta imagen ilustra la región de núcleo accumbens astrocitos que se transfectaron siguiendo el método anterior. Los datos se reimprimieron de Bull et al. 13 con permiso del titular de los derechos de autor. Se pueden encontrar detalles en dicha publicación y el material complementario que acompaña.blank &quot;&gt; Haga clic aquí para ver una versión más grande de esta figura. <img alt="Figura 4" src="/files/ftp_upload/53018/53018fig4.jpg" /> Figura 4. La motivación de las ratas a etanol autoadministrarse se redujo mediante la activación de un transgén que había sobre-expresa en núcleo accumbens astrocitos centrales. Virus se microinyectó y motivación mide a través de punto de interrupción después de permitir una semana para el virus de expresar. El transgén expresó fue un diseñador Receptor Exclusivamente activado por las drogas de diseño (DREADDs). Un ANOVA de una vía (F (2,30) = 3,29, p = 0,04) seguido de un post-hoc de Scheffé revelado que la activación de la DREADD por la administración sistémica de clozapina-N-óxido (CNO, 3 mg / kg, ip ) redujo significativamente la motivación de las ratas a etanol auto-administrarse después de la abstinencia comparación con el vehículo CNO no tuvo ningún efecto en una cohorte igualmente capacitado que expresaba la proteína fluorescente verde (GFP) en lugar del DREADD. Los datos se reimprimieron de Bull et al. 13 con permiso del titular de los derechos de autor. Se pueden encontrar detalles en dicha publicación y el material complementario que acompaña. <img alt="Figura 5" src="/files/ftp_upload/53018/53018fig5.jpg" /> Figura 5. La microinyección de bloqueadores de unión brecha aumentó la motivación de las ratas para autoadministrarse etanol después de la abstinencia Dos bloqueadores brecha cruce fueron evaluados por su efecto sobre la motivación (medido a través de punto de ruptura) de las ratas a autoadministrarse etanol:. Mefloquina y 18- alfa-ácido glicirretínico (18-α). Un ANOVA de dos vías reveló que la microinyección de los bloqueadores de brecha de la salida en el núcleo accumbens de núcleo aumentó la motivación de ratas para autoadministrarse etanol después de la abstinencia (F (3,40) = 5.56, p = 0.003). Los datos se reimprimieron de Bull et al. 13 con permiso del titular de los derechos de autor. Se pueden encontrar detalles en esa publicación yel material complementario que acompaña.

Watch this Scientific Journal Video about Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior at JoVE.com

Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior

Rodents are an appropriate model to investigate the molecular substrates of behavior and complex psychiatric disorders. Brain microinjection in awake rodents can be used to elucidate disease substrates. An efficient and customizable brain microinjection method as well as the execution of an operant paradigm that quantifies motivation is presented.

Roedor cerebro microinyección para el Estudio de los sustratos moleculares de Comportamiento Motivado

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as ...

rodent-brain-microinjection-to-study-molecular-substrates-motivated

Research

JoVE Journal

Behavior

14.2K vistas.  Virginia Commonwealth University School of Medicine. Rodents are an appropriate model to investigate the molecular substrates of behavior and complex psychiatric disorders. Brain microinjection in awake rodents can be used to elucidate disease substrates. An efficient and customizable brain microinjection method as well as the execution of an operant paradigm that quantifies motivation is presented. 

Video: Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior

Stereotaxic Microinjection of Viral Vectors Expressing Cre Recombinase to Study the Role of Target Genes in Cocaine Conditioned Place Preference

Microinjecting recombinant adenoassociated viral (rAAV) vectors expressing Cre recombinase into distinct mouse brain regions to selectively knockout genes of interest allows for enhanced temporally- and regionally-specific control of gene deletion, compared to existing methods. While conditional deletion can also be achieved by mating mice that express Cre recombinase under the control of specific gene promoters with mice carrying a floxed gene, stereotaxic microinjection allows for targeting of discrete brain areas at experimenter-determined time points of interest. In the context of cocaine conditioned place preference, and other cocaine behavioral paradigms such as self-administration or psychomotor sensitization that can involve withdrawal, extinction and/or reinstatement phases, this technique is particularly useful in exploring the unique contribution of target genes to these distinct phases of behavioral models of cocaine-induced plasticity. Specifically, this technique allows for selective ablation of target genes during discrete phases of a behavior to test their contribution to the behavior across time. Ultimately, this understanding allows for more targeted therapeutics that are best able to address the most potent risk factors that present themselves during each phase of addictive behavior.

This article describes how to microinject viral vectors into mouse brain and then test in a conditioned place preference paradigm that includes an acquisition, extinction and reinstatement phase.

La microinyección estereotáxica de vectores virales que expresan Cre recombinasa para estudiar el papel de los genes diana de cocaína acondicionado Place Preferencia

Microinjecting recombinant adenoassociated viral (rAAV) vectors expressing Cre recombinase into distinct mouse brain regions to selectively knockout genes ...

Investigación

Comportamiento

Uncovering Beat Deafness: Detecting Rhythm Disorders with Synchronized Finger Tapping and Perceptual Timing Tasks

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here with the goal of uncovering rhythm disorders, such as beat deafness. Beat deafness is characterized by poor performance in perceiving durations in auditory rhythmic patterns or poor synchronization of movement with auditory rhythms (e.g., with musical beats). These tasks include the synchronization of finger tapping to the beat of simple and complex auditory stimuli and the detection of rhythmic irregularities (anisochrony detection task) embedded in the same stimuli. These tests, which are easy to administer, include an assessment of both perceptual and sensorimotor timing abilities under different conditions (e.g., beat rates and types of auditory material) and are based on the same auditory stimuli, ranging from a simple metronome to a complex musical excerpt. The analysis of synchronized tapping data is performed with circular statistics, which provide reliable measures of synchronization accuracy (e.g., the difference between the timing of the taps and the timing of the pacing stimuli) and consistency. Circular statistics on tapping data are particularly well-suited for detecting individual differences in the general population. Synchronized tapping and anisochrony detection are sensitive measures for identifying profiles of rhythm disorders and have been used with success to uncover cases of poor synchronization with spared perceptual timing. This systematic assessment of perceptual and sensorimotor timing can be extended to populations of patients with brain damage, neurodegenerative diseases (e.g., Parkinson&#8217;s disease), and developmental disorders (e.g., Attention Deficit Hyperactivity Disorder).

Behavioral tasks that allow for the assessment of perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) are presented. Synchronization of finger tapping to the beat of an auditory stimuli and detecting rhythmic irregularities provides a means of uncovering rhythm disorders.

El descubrimiento de golpe Sordera: Trastornos Detección de ritmo con los dedos sincronizada Tapping y tareas de sincronización de percepción

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here ...

A Dual Task Procedure Combined with Rapid Serial Visual Presentation to Test Attentional Blink for Nontargets

When viewers search for targets in a rapid serial visual presentation (RSVP) stream, if two targets are presented within about 500 msec of each other, the first target may be easy to spot but the second is likely to be missed. This phenomenon of attentional blink (AB) has been widely studied to probe the temporal capacity of attention for detecting visual targets. However, with the typical procedure of AB experiments, it is not possible to examine how the processing of non-target items in RSVP may be affected by attention. This paper describes a novel dual task procedure combined with RSVP to test effects of AB for nontargets at varied stimulus onset asynchronies (SOAs). In an exemplar experiment, a target category was first displayed, followed by a sequence of 8 nouns. If one of the nouns belonged to the target category, participants would respond &#8216;yes&#8217; at the end of the sequence, otherwise participants would respond &#8216;no&#8217;. Two 2-alternative forced choice memory tasks followed the response to determine if participants remembered the words immediately before or after the target, as well as a random word from another part of the sequence. In a second exemplar experiment, the same design was used, except that 1) the memory task was counterbalanced into two groups with SOAs of either 120 or 240 msec and 2) three memory tasks followed the sequence and tested remembrance for nontarget nouns in the sequence that could be anywhere from 3 items prior the target noun position to 3 items following the target noun position. Representative results from a previously published study demonstrate that our procedure can be used to examine divergent effects of attention that not only enhance targets but also suppress nontargets. Here we show results from a representative participant that replicated the previous finding.&#160;

This paper describes a novel dual task procedure combined with Rapid Serial Visual Presentation to look at attentional blink at varied stimulus onset asynchronies. This experimental procedure differed from others in that it tested attentional blink for nontargets.

Una Tarea Procedimiento Dual Combinado con Rapid Presentación Visual Serial a Test atencional Blink para no-objetivos

When viewers search for targets in a rapid serial visual presentation (RSVP) stream, if two targets are presented within about 500 msec of each other, the ...

Using the Activity-based Anorexia Rodent Model to Study the Neurobiological Basis of Anorexia Nervosa

Anorexia nervosa (AN) is a psychiatric illness characterized by excessively restricted caloric intake and abnormally high levels of physical activity. A challenging illness to treat, due to the lack of understanding of the underlying neurobiology, AN has the highest mortality rate among psychiatric illnesses. To address this need, neuroscientists are using an animal model to study how neural circuits may contribute toward vulnerability to AN and may be affected by AN. Activity-based anorexia (ABA) is a bio-behavioral phenomenon described in rodents that models the key symptoms of anorexia nervosa. When rodents with free access to voluntary exercise on a running wheel experience food restriction, they become hyperactive &#8211; running more than animals with free access to food. Here, we describe the procedures by which ABA is induced in adolescent female C57BL/6 mice. On postnatal day 36 (P36), the animal is housed with access to voluntary exercise on a running wheel. After 4 days of acclimation to the running wheel, on P40, all food is removed from the cage. For the next 3 days, food is returned to the cage (allowing animals free food access) for 2 hr daily. After the fourth day of food restriction, free access to food is returned and the running wheel is removed from the cage to allow the animals to recover. Continuous multi-day analysis of running wheel activity shows that mice become hyperactive within 24 hr following the onset of food restriction. The mice run even during the limited time during which they have access to food. Additionally, the circadian pattern of wheel running becomes disrupted by the experience of food restriction. We have been able to correlate neurobiological changes with various aspects of the animals&#8217; wheel running behavior to implicate particular brain regions and neurochemical changes with resilience and vulnerability to food-restriction induced hyperactivity.

Here we present a protocol to induce activity-based anorexia (ABA) in female adolescent mice. ABA is a condition of hyperactivity evoked by imposing food restriction on rodents with access to a running wheel. This phenomenon is being used as a model to study the underlying neurobiology of anorexia nervosa.

Utilizando la anorexia Roedor modelo basado en actividades para el Estudio de las bases neurobiológicas de la anorexia nerviosa

Anorexia nervosa (AN) is a psychiatric illness characterized by excessively restricted caloric intake and abnormally high levels of physical activity. A ...

Combined Shuttle-Box Training with Electrophysiological Cortex Recording and Stimulation as a Tool to Study Perception and Learning

Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the necessary equipment is now available by several commercial companies. This protocol provides a detailed description of a two-way shuttle-box avoidance learning paradigm in rodents (here Mongolian gerbils; Meriones unguiculatus) in combination with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical electrophysiological in vivo recordings. The detailed protocol is applicable to study multiple aspects of learning behavior and perception in different rodent species.
Site-specific ICMS of auditory cortical circuits as conditioned stimuli here is used as a tool to test the perceptual relevance of specific afferent, efferent and intracortical connections. Distinct activation patterns can be evoked by using different stimulation electrode arrays for local, layer-dependent ICMS or distant ICMS sites. Utilizing behavioral signal detection analysis it can be determined which stimulation strategy is most effective for eliciting a behaviorally detectable and salient signal. Further, parallel multichannel-recordings using different electrode designs (surface electrodes, depth electrodes, etc.) allow for investigating neuronal observables over the time course of such learning processes. It will be discussed how changes of the behavioral design can increase the cognitive complexity (e.g. detection, discrimination, reversal learning).

Shuttle-box avoidance learning is well-established in behavioral neuroscience. This protocol describes how shuttle-box learning in rodents can be combined with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical in vivo recordings as a tool to study multiple aspects of learning and perception.

Combinado de traslado-Box Entrenamiento con electrofisiológico corteza Grabación y estimulación como una herramienta para estudiar la percepción y aprendizaje

Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

La prueba de roedores psicomotor Vigilancia (rPVT): Un método para evaluar el rendimiento neuroconductual en ratas y ratones

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

Reversible Cooling-induced Deactivations to Study Cortical Contributions to Obstacle Memory in the Walking Cat

On complex, naturalistic terrain, sensory information about an environmental obstacle can be used to rapidly adjust locomotor movements for avoidance. For example, in the cat, visual information about an impending obstacle can modulate stepping for avoidance. Locomotor adaptation can also occur independent of vision, as sudden tactile inputs to the leg by an expected obstacle can modify the stepping of all four legs for avoidance. Such complex locomotor coordination involves supraspinal structures, such as the parietal cortex. This protocol describes the use of reversible, cooling-induced cortical deactivation to assess parietal cortex contributions to memory-guided obstacle locomotion in the cat. Small cooling loops, known as cryoloops, are specially shaped to deactivate discrete regions of interest to assess their contributions to an overt behavior. Such methods have been used to elucidate the role of parietal area 5 in memory-guided obstacle avoidance in the cat.

Complex locomotion in naturalistic environments requiring careful coordination of the limbs involves regions of the parietal cortex. The following protocol describes the use of reversible cooling-induced deactivation to demonstrate the role of parietal area 5 in memory-guided obstacle avoidance in the walking cat.

Desactivaciones reversibles inducida por enfriamiento a estudiar contribuciones corticales a la memoria de obstáculo en el gato caminando

On complex, naturalistic terrain, sensory information about an environmental obstacle can be used to rapidly adjust locomotor movements for avoidance. For ...

Central and Divided Visual Field Presentation of Emotional Images to Measure Hemispheric Differences in Motivated Attention

Two dominant theories on lateralized processing of emotional information exist in the literature. One theory posits that unpleasant emotions are processed by right frontal regions, while pleasant emotions are processed by left frontal regions. The other theory posits that the right hemisphere is more specialized for the processing of emotional information overall, particularly in posterior regions.
Assessing the different roles of the cerebral hemispheres in processing emotional information can be difficult without the use of neuroimaging methodologies, which are not accessible or affordable to all scientists. Divided visual field presentation of stimuli can allow for the investigation of lateralized processing of information without the use of neuroimaging technology. 
This study compared central versus divided visual field presentations of emotional images to assess differences in motivated attention between the two hemispheres. The late positive potential (LPP) was recorded using electroencephalography (EEG) and event-related potentials (ERPs) methodologies to assess motivated attention. Future work will pair this paradigm with a more active behavioral task to explore the behavioral impacts on the attentional differences found.

This study compared central versus divided visual field presentations of emotional images to assess differences in motivated attention between the two hemispheres. The late positive potential (LPP) was recorded using electroencephalography (EEG) and event-related potentials (ERPs) methodologies to assess motivated attention.

Presentación de campo Visual central y se divide de imágenes emocionales para medir diferencias hemisféricas en atención motivado

Two dominant theories on lateralized processing of emotional information exist in the literature. One theory posits that unpleasant emotions are processed ...

Combining Quantitative Food-intake Assays and Forcibly Activating Neurons to Study Appetite in Drosophila

Food consumption is under the tight control of the brain, which integrates the physiological status, palatability, and nutritional contents of the food, and issues commands to start or stop feeding. Deciphering the processes underlying the decision-making of timely and moderate feeding carries major implications in our understanding of physiological and psychological disorders related to feeding control. Simple, quantitative, and robust methods are required to measure the food ingestion of animals after experimental manipulation, such as forcibly increasing the activities of certain target neurons. Here, we introduced dye-labeling-based feeding assays to facilitate the neurogenetic study of feeding control in adult fruit flies. We review available feeding assays, and then describe our methods step-by-step from setup to analysis, which combine thermogenetic and optogenetic manipulation of neurons controlling feeding motivation with dye-labeled food intake assay. We also discuss the advantages and limitations of our methods, compared with other feeding assays, to help readers choose an appropriate assay.

Combining Quantitative Food-intake Assays and Forcibly Activating Neurons to Study Appetite in Drosophila

Quantitative food-intake assays with dyed food provide a robust and high-throughput means to evaluate feeding motivation. Combining the food consumption assay with thermogenetic and optogenetic screens is a powerful approach to investigate the neural circuits underlying appetite in adult Drosophila melanogaster.

Combinar el análisis cuantitativo de la toma de comida y por la fuerza activando las neuronas para estudiar apetito en Drosophila

Food consumption is under the tight control of the brain, which integrates the physiological status, palatability, and nutritional contents of the food, ...

Stress-Enhanced Fear Learning, a Robust Rodent Model of Post-Traumatic Stress Disorder

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders such as post-traumatic stress disorder (PTSD). To understand the biological mechanisms of fear dysregulation in PTSD, it is important to start with a valid animal model of the disorder. This protocol describes the methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of PTSD, in both rats and mice. SEFL was developed to recapitulate critical aspects of PTSD, including long-term sensitization of fear learning caused by an acute stressor. SEFL uses aspects of Pavlovian fear conditioning but produces a distinct and robust sensitized fear response far greater than normal conditional fear responses. The trauma procedure involves placing a rodent in a conditioning chamber and administering 15 unsignaled shocks randomly distributed over 90 minutes (for rat experiments; for mouse experiments, 10 unsignaled shocks randomly distributed over 60 minutes are used). On day 2, rodents are placed in a novel conditioning context where they receive a single shock; then, on day 3 they are placed back in the same context as on day 2 and tested for changes in freezing levels. Rodents that previously received the trauma display enhanced levels of freezing on the test day compared to those that received no shocks on the first day. Thus, with this model, a single highly stressful experience (the trauma) produces extreme fear of the stimuli associated with the traumatic event.

Here we describe the detailed methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of post-traumatic stress disorder, in both rats and mice. The model utilizes aspects of Pavlovian fear conditioning and freezing as an index of enhanced fear in rodents.

Miedo estrés mayor aprendizaje, un modelo de roedor robusto del trastorno por estrés postraumático

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders ...