Les auteurs tiennent à remercier les bourses salariales, subventions d&#39;équipement et les fonds de fonctionnement des Fonds de la recherche en santé du Québec (FRSQ), les Instituts de recherche en santé du Canada (IRSC), en sciences naturelles et en génie du Canada (CRSNG), et l&#39;Université Concordia Programme des chaires de recherche (CRUC), ainsi que la rétroaction réfléchie sur ce manuscrit du Dr Jane Stewart.

1. Logement des animaux <ol><li> Cage: Afin d&#39;enregistrer l&#39;activité d&#39;une roue d&#39;un rongeur individuel, chaque cage doit accueillir un rongeur unique et une roue d&#39;. Parce que les roues de course peut être considéré comme une forme d&#39;enrichissement, tous les rongeurs dans toute étude devrait avoir un accès similaire à une roue qui tourne. </li><li> Changements Literie: manipulation des animaux ainsi que des changements dans des cages ou de la literie peuvent tous avoir des effets non-photique sur les rythmes circadiens 21-23, oui, des cages avec des mailles-sol sont idéales car elles minimisent le contact avec l&#39;animal. Malgré la disponibilité d&#39;un tel plateau de système, des changements de literie doit être évitée pendant les phases critiques d&#39;une expérience. Les alternatives comprennent l&#39;utilisation des lits de plus longue durée, ce qui permettrait des changements plus rares cages, cages ou changeantes sur un calendrier pseudo-aléatoire. </li><li> Boîtes d&#39;isolement: Les cages devraient être conservés dans des boîtes d&#39;isolement qui sont insonorisée, la lumière-controlled et bien ventilé. En fonction de la taille et de la configuration des boîtiers d&#39;isolation, le nombre de cages dans chaque zone est généralement compris entre 1-8. Lorsque le logement cages multiples dans une boîte d&#39;isolation unique, il faut être conscient que les diverses odeurs et des sons provenant d&#39;animaux autres peuvent avoir des effets confondants sur le comportement circadien des animaux individuels. Pour éviter ces problèmes, il faut essayer de loger une cage d&#39;isolation par boîte. </li><li> Ventilation: débit d&#39;air adéquat est impératif de faire les cases d&#39;un milieu familial confortable pour les rongeurs. Le ventilateur dans chaque zone doit être à capuchon, de manière à empêcher la lumière provenant de l&#39;extérieur de la zone d&#39;atteindre l&#39;intérieur. De plus, les fans pourront généralement éliminer l&#39;air de la boîte et le faire sauter dans la pièce. Petites étanches à la lumière des évents permettent à l&#39;air de pénétrer dans les boîtiers d&#39;isolation à partir de plusieurs points, et contribuer à éviter brises mal à l&#39;aise. Afin de vérifier que la ventilation est adéquate, la température à l&#39;intérieur de la boîte d&#39;isolation (lorsquefermé pendant plusieurs heures, avec des lumières sur) doit être pratiquement identique à la température de la pièce où il est logé. </li><li> Eclairage: l&#39;intensité lumineuse de l&#39;environnement devrait être la même dans toutes les cages. Organiser une lumière unique dans un lieu semblable au-dessus de chaque cage, et de toujours utiliser la même marque / type d&#39;ampoule. Utilisez un éclairage d&#39;intensité modérée (100-300 lux) au niveau de la cage. Eviter les niveaux d&#39;éclairement trop élevés, qui sont plus susceptibles de produire des changements directs sur le comportement attribuables à la lumière plutôt que le système circadien, en soi (masquage, par exemple). </li><li> Darkness / Dim éclairage rouge: S&#39;il est nécessaire de manipuler ou traiter les animaux dans l&#39;obscurité (par exemple dans l&#39;obscurité constante ou la nuit), lunettes de vision nocturne devrait être utilisé. Alternativement, parce que le système circadien est relativement insensible aux longueurs d&#39;onde rouges, dim éclairage rouge peut être utilisé. La lumière rouge spécifique que vous utilisez doit être testé pour s&#39;assurer qu&#39;il ne alter une roue d&#39;activité (masquage par exemple) ou réglez l&#39;horloge circadienne (par exemple produire un décalage de phase). </li></ol> 2. Collecte de données (voir Figure 1 - Configuration matérielle Vitalview) <ol><li> Exécution de roues: Le diamètre et l&#39;ergonomie de la roue de roulement va changer la fréquence d&#39;utilisation 24. Ainsi, utiliser des roues plus petites et plus légères pour les souris, et les grandes roues plus lourdes pour les rats. Lors du lavage et de réinstaller les roues, veiller à ce que les roues sont en mesure de spin libre, ne pas &quot;vaciller&quot;, et que les enregistrements micro-interrupteurs sont activés par chaque tour de roue. </li><li> Micro-interrupteurs: Chaque tour de la roue de roulement doit activer un champ magnétique ou mécanique micro-interrupteur. Informations à partir de la micro-interrupteur est transmise par l&#39;intermédiaire d&#39;un canal unique et enregistrés par un ordinateur qui peut bin les données au cours du temps (par exemple tous les 2, 5, 6, ou 10 min). </li><li> Matériel informatique: Notre running roues enregistrements sont réalisés avec Vitalview, une plateforme matérielle et logicielle développée par Mini Mitter ( <a href="http://www.minimitter.com/vitalview_software.cfm" target="_blank">http://www.minimitter.com/vitalview_software.cfm</a> ). Cependant, il ya des plates-formes d&#39;enregistrement tels que ClockLab, développé par Actimetrics ( <a href="http://www.actimetrics.com/ClockLab/" target="_blank">http://www.actimetrics.com/ClockLab/</a> ). Les deux plates-formes de rassembler des données issues de nombreuses sources à canal unique (par exemple, un seul micro-interrupteur actionné par une seule roue tourne) dans un fichier informatique unique. Les données provenant des canaux individuels peuvent ensuite être représentées graphiquement et analysées séparément à une date ultérieure. </li></ol> 3. Les enregistrements de données <ol><li> Fichiers: Les plates-formes logicielles mentionnées ci-dessus peuvent être utilisées pour séparer les voies simples de sorte que les fichiers individuels sont créés pour chaque enregistrement dans une roue. Ces données sont mieux visualisés graphiquement et avec spécialement designeprogrammes d tels que Actiview (Minimitter, Bend, OR), Circadia ou Clocklab (Actimetrics, Wilmette, IL) qui peuvent tous produire périodogrammes et actograms. Cependant, un seul canal fichiers peuvent également être ouvertes et analysées à l&#39;aide de tableurs généraux tels que Excel (Microsoft, Redmond, WA). </li><li> Calcul du temps du rythme circadien (CT): CT 12 est, par définition, le début de la course roues activité chez les rongeurs nocturnes. Parallèlement à la journée de 24 heures, par convention, un jour circadien est divisé en 24 h circadien. Par conséquent, si le rythme circadien free-running période est de 24 heures 30 min tel que mesuré par l&#39;horloge murale, CT 0 aura lieu environ 12 heures 15 min après CT12. </li></ol>

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W., Robinson, B., Stewart, J., Amir, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+central+and+basolateral+nuclei+of+the+amygdala+exhibit+opposite+diurnal+rhythms+of+expression+of+the+clock+protein+Period2.">The central and basolateral nuclei of the amygdala exhibit opposite diurnal rhythms of expression of the clock protein Period2.</a> Proc. Natl. Acad. Sci. U.S.A. 102, 4180-4184 (2005).</li><li>Amir, S., Lamont, E. 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U.S.A. 101, 5339-5346 (2004).</li><li>Guilding, C., Piggins, H. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Challenging+the+omnipotence+of+the+suprachiasmatic+timekeeper:+are+circadian+oscillators+present+throughout+the+mammalian+brain?.">Challenging the omnipotence of the suprachiasmatic timekeeper: are circadian oscillators present throughout the mammalian brain?.</a> Eur. J. Neurosci. 25, 3195-3216 (2007).</li><li>Boulos, Z., Terman, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Food+availability+and+daily+biological+rhythms.">Food availability and daily biological rhythms.</a> Neurosci. Biobehav. Rev. 4, 119-131 (1980).</li><li>Boulos, Z., Rosenwasser, A. 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S., Yamazaki, S., Menaker, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Is+the+food-entrainable+circadian+oscillator+in+the+digestive+system?.">Is the food-entrainable circadian oscillator in the digestive system?.</a> Genes Brain Behav. 2, 32-39 (2003).</li><li>Hara, R., 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=Restricted+feeding+entrains+liver+clock+without+participation+of+the+suprachiasmatic+nucleus.">Restricted feeding entrains liver clock without participation of the suprachiasmatic nucleus.</a> Genes Cells. 6, 269-278 (2001).</li><li>Damiola, F., 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=Restricted+feeding+uncouples+circadian+oscillators+in+peripheral+tissues+from+the+central+pacemaker+in+the+suprachiasmatic+nucleus.">Restricted feeding uncouples circadian oscillators in peripheral tissues from the central pacemaker in the suprachiasmatic nucleus.</a> Genes Dev. 14, 2950-2961 (2000).</li><li>Mrosovsky, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phase+response+curves+for+social+entrainment.">Phase response curves for social entrainment.</a> J. 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Aucun conflit d&#39;intérêt déclaré.

Surveillance des rythmes d&#39;activité par jour à l&#39;aide des roues de roulement est la méthode la plus couramment utilisée et fiable pour évaluer la sortie de l&#39;horloge circadienne chez les rongeurs nocturnes. Roue marche à l&#39;activité, cependant, n&#39;est qu&#39;un des nombreux aspects du comportement et de la physiologie qui peuvent être surveillés en permanence. Bien que la grande majorité des une roue d&#39;activité a lieu pendant la nuit, plus de 30% de l&#39;éveil totale survient au cours ...

<table border="1"&gt;<tbody&gt;<tr&gt;<td<strong&gt; Nom du réactif</strong</td&gt;<td<strong&gt; Société</strong</td&gt;<td<strongNuméro de catalogue&gt;</strong</td&gt;<td<strong&gt; Commentaires (optionnel)</strong</td&gt;</tr&gt;<tr&gt;<td&gt; Vitalview Carte et logiciels</td&gt;<td&gt; Mini Mitter</td&gt;<td&gt; # 855-0030-00</td&gt;<td&gt; (Bend, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; DP24 Dataport</td&gt;<td&gt; Mini Mitter</td&gt;<td&gt; # 840-0024-00</td&gt;<td&gt; (Bend, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; QA4-Module</td&gt;<td&gt; Mini Mitter</td&gt;<td&gt; # 130-0050-00</td&gt;<td&gt; (Bend, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; Interrupteur magnétique</td&gt;<td&gt; Mini Mitter</td&gt;<td&gt; # 130-0015-00</td&gt;<td&gt; (Bend, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; Assembly C-50 Câble</td&gt;<td&gt; Mini Mitter</td&gt;<td&gt; # 060-0045-10</td&gt;<td&gt; (Bend, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; Rat courir roue</td&gt;<td&gt; Mini Mitter</td&gt;<td&gt; # 640-0700-00</td&gt;<td&gt; (Bend, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; Support Cage et le bac</td&gt;<td&gt; Mini Mitter</td&gt;<td&gt; # 640-0400-00</td&gt;<td&gt; (Bend, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; Utilisable couper cage</td&gt;<td&gt; Mini Mitter</td&gt;<td&gt; # 664-2154-00</td&gt;<td&gt; (Bend, OR, USA)</td&gt;</tr&gt;<tr&gt;<tdRez-de-&gt; Grid pour cage</td&gt;<td&gt; Mini Mitter</td&gt;<td&gt; # 676-2154-00</td&gt;<td&gt; (Bend, OR, USA)</td&gt;</tr&gt;<tr&gt;<tdLe bac à déchets&gt;</td&gt;<td&gt; Mini Mitter</td&gt;<td&gt; # 684-2154-00</td&gt;<td&gt; (Bend, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; Boîtier de lampe</td&gt;<td&gt; Microlites scientifique</td&gt;<td&gt; # R-101</td&gt;<td&gt; (Toronto, ON, Canada)</td&gt;</tr&gt;<tr&gt;<td&gt; 4W Lampes fluorescentes</td&gt;<td&gt; Microlites scientifique</td&gt;<td&gt; # F4T5/CW</td&gt;<td&gt; (Toronto, ON, Canada)</td&gt;</tr&gt;<tr&gt;<td&gt; Isolation des chambres</td&gt;<td&gt; Sur mesure</td&gt;<td</td&gt;<td&gt; 28 &quot;H x 20&quot; W x 28 &quot;D ½&quot; Black mélamine.</td&gt;</tr&gt;</tbody&gt;</table

recording and analysis of circadian rhythms in running-wheel activity in rodents

Lorsque les rongeurs ont accès gratuitement à une roue qui tourne dans leur cage, l&#39;utilisation volontaire de cette roue dépend du moment de la journée 1-5. Rongeurs nocturnes, dont les rats, les hamsters et les souris, sont actifs pendant la nuit et relativement inactif pendant la journée. De nombreuses autres mesures comportementales et physiologiques présentent également des rythmes quotidiens, mais chez les rongeurs, la course roues activité constitue une mesure particulièrement fiable et commode de la sortie de l&#39;horloge circadienne principale, le noyau suprachiasmatique (NSC) de l&#39;hypothalamus. En général, par le biais d&#39;un processus appelé entraînement, le modèle journalier de courir roues activité sera naturellement s&#39;aligner sur l&#39;environnement cycle lumière-obscurité (cycle LD, par exemple 12 heures de lumière: 12 h-foncé). Rythmes circadiens sont cependant générés de manière endogène les modèles de comportement qui présentent une période de 24 heures ~, et persister dans l&#39;obscurité constante. Ainsi, en l&#39;absence d&#39;un cycle LD, l&#39;enregistrement et l&#39;analyse de l&#39;activité dans une roue peutêtre utilisée pour déterminer le temps subjectif-de-jour. Parce que ces rythmes sont dirigés par l&#39;horloge circadienne la subjectivité du temps de la journée est appelée rythme circadien (CT). En revanche, quand un cycle LD est présent, le temps de la journée qui est déterminée par le cycle de l&#39;environnement LD est appelé le temps zeitgeber (ZT). Bien que les rythmes circadiens dans une roue d&#39;activité sont généralement liés à l&#39;horloge 6-8 SCN, oscillateurs circadiens dans de nombreuses autres régions du cerveau et du corps 14.9 pourrait également être impliquée dans la régulation des rythmes d&#39;activité quotidienne. Par exemple, les rythmes quotidiens en nourriture anticipation activité ne nécessite pas le SCN 15,16 et à la place, sont en corrélation avec des changements dans l&#39;activité des extra-SCN oscillateurs 17-20. Ainsi, les enregistrements d&#39;activité en cours d&#39;exécution roues peuvent fournir d&#39;importants renseignements sur le comportement non seulement à propos de la sortie de l&#39;horloge du SCN maître, mais aussi sur l&#39;activité des extra-SCN oscillateurs. Ci-dessous nous descrBIE Le matériel et les méthodes utilisés pour enregistrer, analyser et afficher des rythmes circadiens d&#39;activité locomotrice des rongeurs de laboratoire.

<ol><li> Programmes informatiques: programmes informatiques spécialisés sont généralement utilisés dans la production de actograms et le calcul de la période circadienne. Ces programmes comprennent, mais ne sont pas limités à, Actiview (Minimitter, Bend, OR) et Circadia. </li><li> Actograms: Actograms fournir une illustration graphique des tendances quotidiennes de fonctionnement roues activité. Il ya un seul tracé (axe x = 24 h) et double-tracé (axe x = 48 h) actograms. Les deux jours parcelle méth...

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Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

Les rythmes circadiens en cours d&#39;exécution volontaire roues activité chez les mammifères sont étroitement associés aux oscillations moléculaires d&#39;une horloge maître dans le cerveau. En tant que tel, ces rythmes quotidiens du comportement peut être utilisée pour étudier l&#39;influence de facteurs génétiques, pharmacologiques et environnementaux sur le fonctionnement de cette horloge circadienne.

Enregistrement et analyse des rythmes circadiens dans la course roue-activité chez les rongeurs

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

recording-analysis-circadian-rhythms-running-wheel-activity

Research

JoVE Journal

Neuroscience

21.1K vues.  McGill University. Les rythmes circadiens en cours d'exécution volontaire roues activité chez les mammifères sont étroitement associés aux oscillations moléculaires d'une horloge maître dans le cerveau. En tant que tel, ces rythmes quotidiens du comportement peut être utilisée pour étudier l'influence de facteurs génétiques, pharmacologiques et environnementaux sur le fonctionnement de cette horloge circadienne.

Vidéo: Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) pacemakers or clocks. Malfunctions in the human circadian system are associated with numerous diseases or disorders. Much progress towards our understanding of the mechanisms underlying circadian rhythms has emerged from genetic screens whereby an easily measured behavioral rhythm is used as a read-out of clock function. Studies using Drosophila have made seminal contributions to our understanding of the cellular and biochemical bases underlying circadian rhythms. The standard circadian behavioral read-out measured in Drosophila is locomotor activity. In general, the monitoring system involves specially designed devices that can measure the locomotor movement of Drosophila. These devices are housed in environmentally controlled incubators located in a darkroom and are based on using the interruption of a beam of infrared light to record the locomotor activity of individual flies contained inside small tubes. When measured over many days, Drosophila exhibit daily cycles of activity and inactivity, a behavioral rhythm that is governed by the animal's endogenous circadian system. The overall procedure has been simplified with the advent of commercially available locomotor activity monitoring devices and the development of software programs for data analysis. We use the system from Trikinetics Inc., which is the procedure described here and is currently the most popular system used worldwide. More recently, the same monitoring devices have been used to study sleep behavior in Drosophila. Because the daily wake-sleep cycles of many flies can be measured simultaneously and only 1 to 2 weeks worth of continuous locomotor activity data is usually sufficient, this system is ideal for large-scale screens to identify Drosophila manifesting altered circadian or sleep properties.

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

We describe procedures for recording daily locomotor activity rhythms of Drosophila and subsequent data analysis. Locomotor activity rhythms are a reliable behavioral output of animal circadian clocks and are used as the standard readout of clock function when studying circadian mutants or examining how the environment regulates the circadian system.

Dosage activité locomotrice d&#39;étudier les rythmes circadiens et le sommeil dans les paramètres Drosophile</em

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) ...

Recherche

Neurosciences

Measuring Circadian and Acute Light Responses in Mice using Wheel Running Activity

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2. They are responsible for timing our sleep/wake cycles and hormone secretion. Since this timing is not precisely 24-hours, it is synchronized to the solar day by light input. This is accomplished via photic input from the retina to the suprachiasmatic nucleus (SCN) which serves as the master pacemaker synchronizing peripheral clocks in other regions of the brain and peripheral tissues to the environmental light dark cycle3-7. The alignment of rhythms to this environmental light dark cycle organizes particular physiological events to the correct temporal niche, which is crucial for survival8. For example, mice sleep during the day and are active at night. This ability to consolidate activity to either the light or dark portion of the day is referred to as circadian photoentrainment and requires light input to the circadian clock9. Activity of mice at night is robust particularly in the presence of a running wheel. Measuring this behavior is a minimally invasive method that can be used to evaluate the functionality of the circadian system as well as light input to this system. Methods that will covered here are used to examine the circadian clock, light input to this system, as well as the direct influence of light on wheel running behavior.

This article will review methods that can be used to determine circadian function and light responsiveness in mice.

Mesurant les réponses Lumière circadiens et aiguë chez la souris en utilisant l&#39;activité roue qui tourne

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2.  They ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects1-11. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design.
Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (<a href="http://www.neuronexustech.com/" >http://www.neuronexustech.com/</a>; <a href="http://www.sbmicrosystems.com/">http://www.sbmicrosystems.com/</a>; <a href="http://www.acreo.se/">http://www.acreo.se/</a>) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.

We describe methods for large-scale recording of multiple single units and local field potential in behaving rodents with silicon probes. Drive fabrication, probe attachment to the drive and probe implantation processes are illustrated in sufficient details for easy replication.

À grande échelle d&#39;enregistrement des neurones par des sondes de silicium mobiles à se comporter Rongeurs

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

A Fully Automated and Highly Versatile System for Testing Multi-cognitive Functions and Recording Neuronal Activities in Rodents

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be investigated in a single task sequence. The unique feature of this system is a custom-made, acoustically transparent chamber that eliminates many of the issues associated with auditory cue control in most commercially available chambers. The ease with which operant devices can be added or replaced makes this system quite versatile, allowing for the implementation of a variety of auditory, visual, and olfactory behavioral tasks. Automation of the system allows fine temporal (10 ms) control and precise time-stamping of each event in a predesigned behavioral sequence. When combined with a multi-channel electrophysiology recording system, multiple cognitive brain functions, such as motivation, attention, decision-making, patience, and rewards, can be examined sequentially or independently.

In this report, we present a fully automated and highly versatile system capable of simultaneously testing multi-cognitive behaviors and recording neuronal activities for rodents.

Un système entièrement automatisé et hautement polyvalent pour Test Multi-Fonctions cognitives et l&#39;enregistrement des activités neuronales chez les rongeurs

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be ...

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from individual neurons are critical for understanding how neural circuits function under natural conditions. Traditionally, these recordings have been performed 'blind', meaning the identity of the recorded cell is unknown at the start of the recording. Cellular identity can be subsequently determined via intracellular1, juxtacellular2 or loose-patch3 iontophoresis of dye, but these recordings cannot be pre-targeted to specific neurons in regions with functionally heterogeneous cell types. Fluorescent proteins can be expressed in a cell-type specific manner permitting visually-guided single-cell electrophysiology4-6. However, there are many model systems for which these genetic tools are not available. Even in genetically accessible model systems, the desired promoter may be unknown or genetically homogenous neurons may have varying projection patterns. Similarly, viral vectors have been used to label specific subgroups of projection neurons7, but use of this method is limited by toxicity and lack of trans-synaptic specificity. Thus, additional techniques that offer specific pre-visualization to record from identified single neurons in vivo are needed. Pre-visualization of the target neuron is particularly useful for challenging recording conditions, for which classical single-cell recordings are often prohibitively difficult8-11. The novel technique described in this paper uses retrograde transport of a fluorescent dye applied using tungsten needles to rapidly and selectively label a specific subset of cells within a particular brain region based on their unique axonal projections, thereby providing a visual cue to obtain targeted electrophysiological recordings from identified neurons in an intact circuit within a vertebrate CNS.
The most significant novel advancement of our method is the use of fluorescent labeling to target specific cell types in a non-genetically accessible model system. Weakly electric fish are an excellent model system for studying neural circuits in awake, behaving animals12. We utilized this technique to study sensory processing by "small cells" in the anterior exterolateral nucleus (ELa) of weakly electric mormyrid fish. "Small cells" are hypothesized to be time comparator neurons important for detecting submillisecond differences in the arrival times of presynaptic spikes13. However, anatomical features such as dense myelin, engulfing synapses, and small cell bodies have made it extremely difficult to record from these cells using traditional methods11, 14. Here we demonstrate that our novel method selectively labels these cells in 28% of preparations, allowing for reliable, robust recordings and characterization of responses to electrosensory stimulation.

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

Marquage fluorescent rétrograde Permet ciblée extracellulaire seule unité d&#39;enregistrement de neurones identifiés<em&gt; In vivo</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

Méthodes d&#39;enregistrement multi-unités pour caractériser l&#39;activité neuronale dans le Criquet (<em&gt; Schistocerca americana</em&gt; Circuits olfactifs)

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for using blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) to image rodents with simultaneous DBS. DBS fMRI presents a number of technical challenges, including accuracy of electrode implantation, MR artifacts created by the electrode, choice of anesthesia and paralytic to minimize any neuronal effects while simultaneously eliminating animal motion, and maintenance of physiological parameters, deviation from which can confound the BOLD signal.&#160;Our laboratory has developed a set of procedures that are capable of overcoming most of these possible issues. For electrical stimulation, a homemade tungsten bipolar microelectrode is used, inserted stereotactically at the stimulation site in the anesthetized subject. In preparation for imaging, rodents are fixed on a plastic headpiece and transferred to the magnet bore. For sedation and paralysis during scanning, a cocktail of dexmedetomidine and pancuronium is continuously infused, along with a minimal dose of isoflurane; this preparation minimizes the BOLD ceiling effect of volatile anesthetics. In this example experiment, stimulation of the subthalamic nucleus (STN) produces BOLD responses which are observed primarily in ipsilateral cortical regions, centered in motor cortex. Simultaneous DBS and fMRI allows the unambiguous modulation of neural circuits dependent on stimulation location and stimulation parameters, and permits observation of neuronal modulations free of regional bias. This technique may be used to explore the downstream effects of modulating neural circuitry at nearly any brain region, with implications for both experimental and clinical DBS.

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

La stimulation cérébrale profonde avec simultanée IRMf chez les rongeurs

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments

Afferent nerves not only convey information concerning normal physiology, but also signal disturbed homeostasis and pathophysiological processes of the different organ systems from the periphery towards the central nervous system. As such, the increased activity or &#39;sensitization&#39; of mesenteric afferent nerves has been allocated an important role in the pathophysiology of visceral hypersensitivity and abdominal pain syndromes.
Mesenteric afferent nerve activity can be measured in vitro in an isolated intestinal segment that is mounted in a purpose-built organ bath and from which the splanchnic nerve is isolated, allowing researchers to directly assess nerve activity adjacent to the gastrointestinal segment. Activity can be recorded at baseline in standardized conditions, during distension of the segment or following the addition of pharmacological compounds delivered intraluminally or serosally. This technique allows the researcher to easily study the effect of drugs targeting the peripheral nervous system in control specimens; besides, it provides crucial information on how neuronal activity is altered during disease. It should be noted however that measuring afferent neuronal firing activity only constitutes one relay station in the complex neuronal signaling cascade, and researchers should bear in mind not to overlook neuronal activity at other levels (e.g., dorsal root ganglia, spinal cord or central nervous system) in order to fully elucidate the complex neuronal physiology in health and disease.
Commonly used applications include the study of neuronal activity in response to the administration of lipopolysaccharide, and the study of afferent nerve activity in animal models of irritable bowel syndrome. In a more translational approach, the isolated mouse intestinal segment can be exposed to colonic supernatants from IBS patients. Furthermore, a modification of this technique has been recently shown to be applicable in human colonic specimens.

In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments

Mesenteric afferent nerves convey information from the gastrointestinal tract towards the brain regarding normal homeostasis as well as pathophysiology. Gastrointestinal afferent nerve activity can be assessed by mounting isolated intestinal segments with attached afferent nerves into an organ bath, isolating the nerve, and assessing basal as well as stimulated activity.

In vitro Enregistrement de mésentérique Afférent activité nerveuse chez la souris jéjunale et Segments colique

Afferent nerves not only convey information concerning normal physiology, but also signal disturbed homeostasis and pathophysiological processes of the ...

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic tectum, is a popular model to study how neural circuits self-assemble. The ability to carry out whole cell patch clamp recordings from tectal neurons and to record RGC-evoked responses, either in vivo or using a whole brain preparation, has generated a large body of high-resolution data about the mechanisms underlying normal, and abnormal, circuit formation and function. Here we describe how to perform the in vivo preparation, the original whole brain preparation, and a more recently developed horizontal brain slice preparation for obtaining whole cell patch clamp recordings from tectal neurons. Each preparation has unique experimental advantages. The in vivo preparation enables the recording of the direct response of tectal neurons to visual stimuli projected onto the eye. The whole brain preparation allows for the RGC axons to be activated in a highly controlled manner, and the horizontal brain slice preparation allows recording from across all layers of the tectum.

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

In this paper, we discuss three brain preparations used for whole cell patch clamp recording to study the retinotectal circuit of Xenopus laevis tadpoles. Each preparation, with its own specific advantages, contributes to the experimental tractability of the Xenopus tadpole as a model to study neural circuit function.

Préparations et protocoles pour toute cellule Patch Clamp enregistrement des neurones Tectal Xenopus laevis

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in neurocognitive disorders. Despite the popularization of prepulse inhibition (PPI) in recent years, many current protocols promote using a percent of control measure, thereby precluding the assessment of temporal processing. The present study used cross-modal PPI and gap prepulse inhibition (gap-PPI) to demonstrate the benefits of employing a range of interstimulus intervals (ISIs) to delineate effects of sensory modality, psychostimulant exposure, and age. Assessment of sensory modality, psychostimulant exposure, and age reveals the utility of an approach varying the interstimulus interval (ISI) to establish the shape of the ISI function, including increases (sharper curve inflections) or decreases (flattening of the response amplitude curve) in startle amplitude. Additionally, shifts in peak response inhibition, suggestive of a differential sensitivity to the manipulation of ISI, are often revealed. Thus, the systematic manipulation of ISI affords a critical opportunity to evaluate temporal processing, which may reveal the underlying neural mechanisms involved in neurocognitive disorders.

Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.