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.

La procédure présentée ici est un moyen efficace pour la fabrication de micro-injection canules et microinjecteurs qui aideront à élucider les substrats moléculaires de comportement motivé. Cette méthode offre plusieurs avantages. D&#39;abord, par la fabrication propres implants et microinjecteurs de un, de nouveaux paramètres expérimentaux peuvent être rapidement optimisées;-à-dire, on n&#39;a pas besoin d&#39;attendre pour les composants sur mesure pour arriver. Deuxièmement, en raison du faible diamètre de la canule, plus canules peuvent être implantés simultanément. Cela raccourcit le temps chirurgical nécessaire, ce qui peut améliorer la survie, et permet également de multiples implants par animal. Troisièmement, le logiciel utilisé pour contrôler les chambres opératoires accueille facilement les horaires de rapport progressistes depuis un ratio paradigme fixe peut être rapidement converti en un ratio paradigme progressive en appliquant simplement une liste de paramètres de transition de l&#39;événement qui contient le programme de renforcement souhaitée. Êtrelargement utile, un procédé de micro-injection a été présenté générique qui devrait être largement applicable pour la micro-injection de presque n&#39;importe quel réactif actuellement disponibles. Par conséquent, nous prévoyons que cette technique continuera d&#39;être d&#39;une grande utilité similaire à l&#39;avenir avec des modifications mineures. En changeant seulement quelques variables, cette approche peut être appliquée à un grand nombre de réactifs. Les paramètres qui le plus souvent être manipulées comprennent la longueur du micro-injecteur qui fait saillie à partir de la canule, le volume de l&#39;injection et la vitesse d&#39;injection. Par exemple, on peut vouloir l&#39;injecteur pour faire saillie plus loin de la pointe de la canule pour éviter la cicatrice gliale qui forme généralement autour des implants chroniques. En outre, on peut souhaiter pour injecter un volume plus important. Pour les micro-injections de virus du striatum, un volume de 1 ul est habituellement utilisé et ce volume est typiquement injecté sur une période de temps plus longue (souvent 7-10 min plus 3-10 min temps de diffusion supplémentaire) par rapport à l&#39;utilisation queD pour les réactifs pharmacologiques (généralement 0,3 - 0,5 pi plus de 2 - 3 min plus 1 - 3 min temps de diffusion supplémentaire). L&#39;utilisateur devrait consulter la littérature et / ou empiriquement déterminer les paramètres les mieux adaptés à leurs besoins. Peu importe, le succès de cette procédure dépend essentiellement de 4 variables: 1) la longueur de la canule, 2) la longueur de microinjecteur, 3) la qualité du modèle microinjecteur de pulvérisation, et 4) l&#39;intégrité du système avant l&#39;injection. Parce emplacement de micro-injection est fonction de la profondeur que le micro-injecteur fait saillie à partir de la canule, il est impératif que les deux canules (étape 1.2.8) et la longueur de micro-injecteur (post-flexion, l&#39;étape 2.2.1) sont à la fois connue avec précision et uniforme entre tous les sujets . Cela peut facilement être contrôlé par facilement rejeter tout mettre en œuvre ce ne est pas la longueur nécessaire à la ré-mesure finale. En outre, l&#39;emplacement d&#39;injection ne peut être prédit si elle se produit immédiatement en dessous de la canule de guidage. Ainsi, toute microinjecteur That Does pas vaporiser une longue, mince filet sur des tests (étapes 2.4.6) devraient être rejetées. Une injection de qualité est également liée à l&#39;intégrité du système avant l&#39;injection. Si après la distribution toute l&#39;eau de l&#39;injecteur (avant le remplissage avec le réactif) multiples taches sont observées sur le laboratoire-essuyer, puis une fuite doit être corrigée (Note sur l&#39;étape 2.4.8). En outre, si la bulle (étape 2.4.9) qui sépare le médicament à partir de l&#39;eau dans la tubulure PE20 est pas une, bulle unique (après remplissage de la micro-injecteur ayant réactif), puis l&#39;injecteur est partiellement obstrué. Ce sabot pourrait prévenir ou détourner l&#39;injection. Cela aussi peut être aisément remédié (Note sur l&#39;étape 2.4.8). Si l&#39;on veut micro-injection alors que l&#39;animal est dans le cadre stéréotaxique il ya trois alternatives. En premier lieu, on pourrait augmenter la longueur du col de micro-injecteur de telle sorte qu&#39;il peut être tenu fermement par le manipulateur stéréotaxique et également prolonger suffisamment loin pour permettre la connexion à un tube PE20. D&#39;autre part, sure temporellement pourrait implanter une canule et d&#39;utiliser la norme microinjecteur présenté ici. Troisièmement, on pourrait utiliser dessinée et des pipettes en verre poli. 16,17 Une limitation importante de la procédure présentée ici est qu&#39;il est mieux menée chez le rat bien gérées qui sont familiers avec la procédure. Les rats utilisés pour les données décrites dans la section des résultats nécessaires pas de procédures spéciales de manipulation parce que le même enquêteur manipulé des rats sur une base quotidienne pendant plus de 2 mois. Cela comprenait l&#39;observation quotidienne et la manipulation de l&#39;implant chirurgical pendant au moins 2 semaines. Cependant, les rats peuvent être habitués rapidement par un certain nombre de techniques qui sont utilisées pour avant l&#39;essai d&#39;inhibition de pré-impulsion, qui peut être affecté par le stress. Ces techniques d&#39;habituation spéciales ont été bien détaillé précédemment. 43 En plus de ces procédures, il est conseillé que les rats sont habitués à la procédure de microinjection où microinjecteurs raccourcis sont utilisés duanneau injections «fictives». Au cours de ces injections simulées, il est essentiel que le microinjecteur pas dépasser dans le tissu afin de limiter les dommages aux tissus. En d&#39;autres termes, le micro-injecteur ne doit pas être plié plus de 14 mm. Ainsi, l&#39;accoutumance complet requis pour la mise en oeuvre optimale de cette technique peut être considérée comme une limitation. Bien que plusieurs paradigmes comportementaux existent pour mesurer la motivation, le rapport progressif est couramment utilisé pour quantifier l&#39;effort que l&#39;objet est disposé à exercer pour obtenir un renforcement. Le ratio paradigme progressif produit une mesure connue sous le nom de point d&#39;arrêt, qui est souvent défini comme le nombre maximal de presses à levier dans le dernier rapport complété;..-À-dire, répondre maximale qui a généré un renforçateur 21 Le ratio progressive est sensible à renforçateur grandeur. Par exemple, la cocaïne supérieur (ou saccharose) doses produisent un point d&#39;arrêt supérieur et de la cocaïne inférieure (ou saccharose) doses donnent une breakp inférieureoint. 21,22 conséquent, point d&#39;arrêt est un proxy utilisé couramment pour la motivation et / ou l&#39;efficacité de renforcement. 21,23-26 Parce que l&#39;intention du point d&#39;arrêt est de déterminer quand l&#39;animal cesse de répondre, un paramètre important du rapport progressive paradigme est durée de la session. Longueurs de session finis peuvent mettre un faux plafond sur les valeurs de point d&#39;arrêt, ce qui peut être exacerbée par des pré-traitements qui diminuent anormalement le taux d&#39;auto-administration ou cette augmentation post-renforcement pause. Cette confondre peut être surmonté par un certain nombre d&#39;approches;.. Par exemple, des séances qui se terminent lorsque l&#39;animal a retenu répondre par un multiple de l&#39;intervalle moyen entre perfusion 44 Une variante plus couramment appliquée de cette approche est de mettre fin à des sessions fois répondre a été retenu par une valeur déterminée de façon empirique qui est maintenue constante dans toutes les disciplines. Nous avons fourni la méthode à appliquer cette approche à l&#39;étape 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.

Ici, nous montrons des exemples de résultats qui peuvent être obtenus avec le procédé ci-dessus. Montré premier est un quartier typique qui est transfectées par des vecteurs viraux (Figure 3). En général, un volume de ~ 1 transfectées 3 mm est obtenue dans un tissu striatal. Le volume de la région transfectée peut être quantifiée à travers des sections en série à l&#39;aide de la méthode de Cavalieri 32 Fait important, le volume transfectées dépend de nombreux facteurs tels que le type de tissu étant transfectée. sérotype viral; promoteur; débit et le volume injecté; nombre de particules injectées; injecter pH; et si réactifs hyperosmolaires, tels que le mannitol, ont été utilisés. 33,34 Typiquement, nous micro-injection 10 12 particules / ml, 1 pl / latéraux plus de 10 min, et laissez 7 min supplémentaires avant de remplacer obturateurs. En outre, le produit à injecter est généralement autour de pH 8. Ensuite, nous montrons que la motivation peut être manipulé par soit l&#39;expression du transgène (figure 4)ou une micro-injection de réactif pharmacologique (figure 5). Dans la figure 4, nous avons exprimé un récepteur Designer Exclusivement Activé par Designer Drugs (de DREADDs). La région codante du récepteur a été suivie par DREADD un site d&#39;entrée de ribosome interne et une cassette mCitrine. Le mCitrine permet la visualisation commode de cellules transfectées. Le DREADD a été couplé à la protéine G hétérotrimérique-Gaq. Activation du couplage DREADD Gaq peut stimuler les astrocytes, 28,35 et la DREADD lui-même peut être activé par l&#39;administration systémique de la clozapine N-oxyde (CNO, 3 mg / kg, ip). Les rats ont été formés 14,22,36 à levier répondre pour le renforcement de l&#39;éthanol, où 3 presses à levier ont donné une chance de boire pendant les jours 1 h séances de plus de 60 jours consécutifs. Ensuite, les rats ont été contraints à l&#39;abstinence, et DREADDs Gaq couplés ont été exprimés dans le noyau accumbens astrocytes de base. Après trois semaines d&#39;abstinence, la motivation à l&#39;auto-ADMI<html> nistre de l&#39;éthanol a été mesurée par point d&#39;arrêt. 21,27-30 activation du noyau accumbens astrocytes de base, par l&#39;intermédiaire de l&#39;administration systémique CNO, a diminué la motivation des rats pour reprendre l&#39;éthanol auto-administration après l&#39;abstinence par rapport à un véhicule. Surtout, le CNO n&#39;a eu aucun effet dans une cohorte aussi formé qui exprimait la protéine fluorescente verte au lieu de la DREADD. <img alt="Figure 3" src="/files/ftp_upload/53018/53018fig3.jpg" /> Figure 3. striatum ventral région Représentant transfectées par le virus microinjecté. Cette image illustre la région du noyau accumbens astrocytes qui ont été transfectées en suivant la méthode ci-dessus. Les données sont reproduites à partir de Bull et al. 13 avec la permission du titulaire du droit d&#39;auteur. Les détails peuvent être trouvés dans cette publication et le matériel complémentaire d&#39;accompagnement.blank &quot;&gt; S&#39;il vous plaît cliquer ici pour voir une version plus grande de cette figure. <img alt="Figure 4" src="/files/ftp_upload/53018/53018fig4.jpg" /> Figure 4. Motivation des rats à auto-administrer de l&#39;éthanol a été réduite par l&#39;activation d&#39;un transgène qui a été sur-exprimé dans le noyau accumbens astrocytes de base. Le virus a été microinjecté et la motivation mesurée par point d&#39;arrêt après avoir laissé une semaine pour le virus d&#39;exprimer. Le transgène exprimé était un créateur Receptor Exclusivement Activé par Designer Drugs (de DREADDs). Une ANOVA à sens unique (F (2,30) = 3,29, p = 0,04), suivie par un post-hoc Scheffé révélé que l&#39;activation de la DREADD par l&#39;administration systémique de la clozapine N-oxyde (CNO, 3 mg / kg, ip ) réduit de façon significative la motivation des rats à auto-administrer de l&#39;éthanol après l&#39;abstinence par rapport à un véhicule CNO n&#39;a eu aucun effet dans une cohorte aussi formé qui exprimait la protéine fluorescente verte (GFP) au lieu de la DREADD. Les données sont reproduites à partir de Bull et al. 13 avec la permission du titulaire du droit d&#39;auteur. Les détails peuvent être trouvés dans cette publication et le matériel complémentaire d&#39;accompagnement. <img alt="Figure 5" src="/files/ftp_upload/53018/53018fig5.jpg" /> Figure 5. microinjection de Gap bloqueurs de jonction a augmenté la motivation des rats à l&#39;auto-administrer éthanol après l&#39;abstinence Deux bloquants des jonctions lacunaires ont été évalués pour leur effet sur ​​la motivation (mesurée par l&#39;intermédiaire de points d&#39;arrêt) de rats à l&#39;auto-administrer de l&#39;éthanol:. Méfloquine et 18- a-acide glycyrrhétinique (18-α). Une ANOVA à deux voies a révélé que la micro-injection de bloqueurs écart-jonction dans le noyau accumbens base a augmenté la motivation des rats à l&#39;auto-administrer éthanol après l&#39;abstinence (F (3,40) = 5,56, p = 0,003). Les données sont reproduites à partir de Bull et al. 13 avec la permission du titulaire du droit d&#39;auteur. Les détails peuvent être trouvés dans cette publication etle matériel complémentaire d&#39;accompagnement.

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.

Le cerveau de rongeurs microinjection pour l&#39;étude de substrats moléculaires de Motivé Comportement

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 vues.  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. 

Vidéo: 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 micro-injection stéréotaxique de vecteurs viraux exprimant la recombinase Cre pour étudier le rôle des gènes cibles de cocaïne préférence de place conditionnée

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

Recherche

Sciences du comportement

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.

Découvrir beat Surdité: détection des troubles du rythme avec le doigt taraudage synchronisé et tâches de temporisation perceptuelles

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.

Une procédure de double tâche combinée avec Rapid Serial Visual Presentation tester attentionnel pour Blink organismes non visés

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.

Utilisation de l&#39;anorexie rongeurs modèle basée sur les activités à l&#39;étude des bases neurobiologiques de l&#39;anorexie mentale

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.

Combiné navette-Box formation avec électrophysiologique Cortex enregistrement et la stimulation comme un outil pour étudier perception et l&#39;apprentissage

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.

Le Rongeur psychomoteur Vigilance Test (RPVT): une méthode d&#39;évaluation des performances neurocomportementales chez les rats et les souris

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.

Réversible par refroidissement désactivations pour étudier les Contributions corticales à Obstacle mémoire chez le chat marche

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.

Présentation du champ visuel central et divisée d’Images émotionnelles pour mesurer les différences hémisphériques dans Attention motivée

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.

Combinant les dosages quantitatifs-la prise alimentaire et activation par la force des neurones pour étudier l’appétit chez la drosophile

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.

Peur stress-Enhanced Learning, un modèle robuste rongeur du syndrome de Stress post-traumatique

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