Vorremmo ringraziare Helmut Scheu per l'opportunità di condurre la nostra ricerca nel porcile presso Hofgut Neumühle.

Tutte le procedure sono state approvate dal comitato etico locale (n. 23177-07/G10-1-010/G 15-15-011) e seguirono l'europea e disposizioni nazionali tedesche (Comunità Consiglio direttiva europea 86/609/CEE; Tierschutzgesetz).
Animale tutte le procedure sono state eseguite in conformità con il Medical Center di regolamenti del Comitato Johannes Gutenberg-Universität Mainz cura degli animali.
1. installazione
<ol><li>Prima dell'esperimento, è necessario controllare per eventuali disturbi sulla linea e trovare una posizione adeguata per il set-up e l'antenna. Rumore di linea è visibile come una sinusoide 60 o 50 Hz. Nota: Il posizionamento dell'antenna e soprattutto la distanza tra il trasmettitore e il ricevitore dipende la forza di trasmissione del sistema. Il sistema utilizzato qui è regolabile. È stato registrato a una potenza relativamente bassa, con un raggio di trasmissione circa 3. Inoltre, le recinzioni metalliche in porcile possono smorzare il segnale e causare interferenze. In questo caso, è necessario posizionare l'antenna all'interno della gabbia di metallo.</li>
	<li>Utilizzare un cavo tamburo per fornire il set-up con alimentazione di rete. Collegare il computer portatile, l'unità di ricezione e il convertitore analogico-digitale (se necessario) per il sistema di telemetria specifico che viene utilizzato. Nota: Il sistema di telemetria utilizzato qui inviati dati digitali al ricevitore. Questo potrebbe essere diverso per altri sistemi.</li>
	<li>Posizionare gli elettrodi, i collanti, i Q-tips e le salviettine, come pure i blocchi di miscelazione, su una tabella separata.</li>
	<li>Preparare gli elettrodi con cavi corti. Per effettuare questa operazione, tagliare gli elettrodi e saldarli nuovamente con una durata più breve possibile, a seconda delle dimensioni dell'animale. I cavi devono essere abbastanza a lungo per collegare le posizioni di registrazione desiderata sulla testa con l'unità di EEG telemetrico, trasmissione dei dati. Cavi troppo lunghi devono essere indietreggiò e rivestiti di pelle in silicone adesivo elastomero. Cavi più lunghi che devono essere indietreggiò rendono la patch in silicone più grandi e pesanti.</li>
</ol>2. porcellino
<ol><li>Prendere un maialino afferrando e alla gamba o al torace. Tenerlo ed essere consapevoli di ogni defecazione o la minzione.</li>
	<li>Se necessario, contrassegnare il maialino con un numero.</li>
	<li>Avvolgere il porcellino in un asciugamano. Il maialino si calmerà. Essere consapevoli di surriscaldamento il maialino.</li>
	<li>Tenere il maialino con una mano il corpo o l'avambraccio. Usare l'altra mano per tenere il muso. Essere consapevoli di surriscaldamento il maialino e assicurarsi che sia libera di respirare correttamente.</li>
</ol>3. elettrodi
<ol><li>Avere una seconda persona collegare gli elettrodi.</li>
	<li>Pulire la pelle dallo sporco con acqua o etanolo. Se necessario, radersi la testa.</li>
	<li>Rimuovere le cellule morte della pelle con un gel di EEG abrasivo e un q-tip. Rimuovere il gel abrasivo in seguito. In alternativa, utilizzare carta vetrata.</li>
	<li>Difficoltà elettrodi autoadesivi nella posizione desiderata. Posizionare l'elettrodo di terra sopra il cervelletto (tra le orecchie) e l'elettrodo di riferimento sul naso. Posizionare l'elettrodo di registrazione nella posizione desiderata. Nota: In questo caso, una registrazione unipolare è stata effettuata, perché il riferimento è stato collocato in una posizione neutrale (naso). Non c'è nessun sistema standardizzato disponibile per suinetti fino ad ora. Qui, una posizione di registrazione parietale è stata usata (tra l'occhio e l'orecchio) sull'emisfero del cervello destro.</li>
	<li>Collegare i cavi per l'unità in telemetria. Accendere l'unità. A seconda del sistema di telemetria utilizzato, questo potrebbe essere un interruttore magnetico o un segnale di sveglia radio frequenza.</li>
	<li>Coprire l'unità di telemetria e tutti i cavi così come tutti gli elettrodi con gomma di silicone adesivo bicomponente pelle (Vedi Tabella materiali). Mescolando quantità uguali di entrambi i componenti, il tempo di polimerizzazione sarà nella gamma di 1 min. occhi e ciglia non devono essere coperto con la gomma.</li>
	<li>Attendere che la gomma di silicone è completamente guarita.</li>
	<li>Riposizionare il porcellino nel porcile.</li>
	<li>Osservare il maialino per vedere se sta mostrando segni di disagio su un periodo più lungo di tempo (alcuni minuti).</li>
</ol>4. misurazione
<ol><li>Attendere che il maialino ha recuperato e inizia a sincronizzare il comportamento con quello dei suoi fratelli (alimentazione, giocare, dormire), di solito dopo 30 s (Figura 1).</li>
	<li>Attendere per fasi di sonno, se lo si desidera. Il tempo di registrazione dipende la questione scientifica specifica. Qui, sono state usate le sessioni di registrazione di 10 min.</li>
	<li>Se l'unità in telemetria è coperto da più di 2 altri suinetti, il segnale potrebbe essere troppo basso per il ricevitore. Spingere delicatamente i suinetti distanza se stanno dormendo sulla parte superiore. Essere consapevoli della scrofa; esso potrebbe reagire aggressivamente.</li>
	<li>Avviare la registrazione con il software di acquisizione dati (Vedi Tabella materiali).</li>
</ol>5. finitura
<ol><li>Dopo la registrazione (di solito diverse ore), prendere nuovamente il maialino come è stato descritto nel passaggio 2. Essere consapevoli della scrofa; esso potrebbe reagire aggressivamente.</li>
	<li>Sollevare delicatamente la gomma di silicone a un bordo. Quindi, rimuovere la patch intera della gomma di silicone contenente gli elettrodi e l'unità in telemetria. Siate cauti con gli occhi di porcellino.</li>
	<li>Riposizionare il porcellino nel porcile.</li>
</ol>

<ol>
	<li>Conrad, M. S., Sutton, B. P., Dilger, R. N., Johnson, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+vivo+three-dimensional+magnetic+resonance+imaging-based+averaged+brain+collection+of+the+neonatal+piglet+(Sus+scrofa).">An in vivo three-dimensional magnetic resonance imaging-based averaged brain collection of the neonatal piglet (Sus scrofa).</a> PLoS ONE. 9 (9), e107650 (2014).</li><li>de Camp, N. V., Hense, F., Lecher, B., Scheu, H., Bergeler, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+for+preterm+cortical+development+using+non+invasive+clinical+EEG.">Models for preterm cortical development using non invasive clinical EEG.</a> Translational Neuroscience. 8, 211-224 (2017).</li><li>Lapray, D., Bergeler, J., Dupont, E., Thews, O., Luhmann, H. J., Barculo, D., Daniels, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+telemetric+system+for+recording+brain+activity+in+small+animals.">A novel telemetric system for recording brain activity in small animals.</a> Telemetry: Research, Technology and Applications. , 195-203 (2009).</li><li>Kim, D., Yeon, C., Kim, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+experimental+validation+of+a+dry+non-+invasive+multi-channel+mouse+scalp+EEG+sensor+through+visual+evoked+potential+recordings.">Development and experimental validation of a dry non- invasive multi-channel mouse scalp EEG sensor through visual evoked potential recordings.</a> Sensors. 17, 326 (2017).</li><li>Moshayedi, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationship+between+glial+cell+mechanosensitivity+and+foreign+body+reactions+in+the+central+nervous+system.">The relationship between glial cell mechanosensitivity and foreign body reactions in the central nervous system.</a> Biomaterials. 35, 3919-3925 (2014).</li><li>Barrese, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Failure+mode+analysis+of+silicon-based+intracortical+microelectrode+arrays+in+non-human+primates.">Failure mode analysis of silicon-based intracortical microelectrode arrays in non-human primates.</a> Journal of Neural Engineering. 10, 066014 (2013).</li><li>Hellstr&#246;m-Westas, L., Ros&#233;n, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electroencephalography+and+brain+damage+in+preterm+infants.">Electroencephalography and brain damage in preterm infants.</a> Early Human Development. 81, 255-261 (2005).</li><li>Lloyd, R. O., Goulding, R. M., Filan, P. M., Boylan, G. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Overcoming+the+practical+challenges+of+electroencephalography+for+very+preterm+infants+in+the+neonatal+intensive+care+unit.">Overcoming the practical challenges of electroencephalography for very preterm infants in the neonatal intensive care unit.</a> Acta Paediatrica. , 152-157 (2015).</li><li>Clancy, B., Finlay, B. L., Darlington, R. B., Anand, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extrapolating+brain+development+from+experimental+species+to+humans.">Extrapolating brain development from experimental species to humans.</a> Neurotoxicology. 28, 931-937 (2007).</li><li>Iyer, K. K., 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=Cortical+burst+dynamics+predict+clinical+outcome+early+in+extremely+preterm+infants.">Cortical burst dynamics predict clinical outcome early in extremely preterm infants.</a> Brain. 138, 2206-2218 (2015).</li><li>Luhmann, H., de Camp, N., Bergeler, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+brain+activity+in+preterms:+mathematics+helps+to+predict+clinical+outcome.">Monitoring brain activity in preterms: mathematics helps to predict clinical outcome.</a> Brain. 138, 2114-2125 (2015).</li><li>Dragomir, A., Akay, Y., Curran, A. K., Akay, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complexity+measures+of+the+central+respiratory+networks+during+wakefulness+and+sleep.">Complexity measures of the central respiratory networks during wakefulness and sleep.</a> Journal of Neural Engineering. 5, 254-261 (2008).</li><li>Peever, J., Fuller, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biology+of+REM+sleep.">The biology of REM sleep.</a> Current Biology. 27, R1237-R1248 (2017).</li><li>Robert, S., Dallaire, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polygraphic+Analysis+of+the+sleep-wake+states+and+the+REM+Sleep+periodicity+in+domesticated+pigs+(Sus+scrofa).">Polygraphic Analysis of the sleep-wake states and the REM Sleep periodicity in domesticated pigs (Sus scrofa).</a> Physiology &amp; Behavior. 37 (2), 289-293 (1986).</li></ol>

Gli autori non hanno nulla a rivelare.

Un passo fondamentale nel protocollo è il contatto di pelle adeguata con gli elettrodi, soprattutto l'elettrodo di terra, per realizzare registrazioni stabile con a basso rumore. Inoltre, poiché i suinetti sono molto agili, è importante coprire l'intero sistema con gomma di silicone per proteggere gli elettrodi e l'unità in telemetria. Inoltre, se gli esperimenti vengono condotti in una stalla con un pavimento a doghe, essere cauti con dispositivi di piccole dimensioni o connettori.
In cas...

Suinetti sono un sistema di modello emergente per neuroscienza1. Al fine di rafforzare la ricerca traslazionale, abbiamo inventato un metodo per registrare EEGs non invasiva, clinica da sfrenato suinetti2 (Figura 1 e Figura 2). Due prerequisiti per un uso traslazionale di registrazioni di EEG, per quanto riguarda i modelli di EEG connessi con maturazione corticale, sono una metodologia non invasiva, paragonabile alla regolazione clinica e l'astinenza di sedativi o anestesia. Un canale telemetria system3 in combinazione con elettrodi autoadesivi può essere fissato in circa 5 min. in seguito, i suinetti saranno recuperare rapidamente dalla procedura di gestione e sincronizzare la loro alimentazione e sonno comportamento a quello degli altri la scrofa e porcellini.
Anche se ci sono già i tentativi di utilizzare le registrazioni di EEG non invasivo da animali sedati4, maggior parte dei studi di elettroencefalografia da animali sono condotti con approcci invasivi. Questi metodi hanno effetti collaterali per quanto riguarda i processi infiammatori intorno al5,di elettrodi impiantati6 e, nella maggior parte dei casi, richiedono una separazione sociale degli animali a causa di componenti esterni del sistema EEG impiantato. Quindi, la traduzione di questi dati nel contesto clinico è difficile. La necessità di approcci traslazionale sta diventando chiaro dal fatto che non è ancora noto come una maturazione "normale" del cervello durante lo sviluppo corticale iniziale è rappresentata da elettroencefalografia clinica, non-invasivo7. Questo divario di conoscenza è causato da problemi tecnici connessi con le registrazioni di EEG da pretermine babies8. Nei sistemi di modello animali, modelli di sviluppo corticale precoce meglio sono accessibili, poiché la maggior parte degli animali sono nati con un "cervello pretermine" rispetto umano sviluppo corticale9. Oltre a modelli conservati dello sviluppo corticale attraverso specie2, recentemente è stato dimostrato che le registrazioni di EEG da bambini prematuri possono anche prevedere il risultato clinico individuale durante la successiva vita10,11. Il metodo qui descritto è particolarmente utile per gli aspetti traslazionali della neuroscienza inerente allo sviluppo.

<table border="0" cellpadding="0" cellspacing="0" width="867">
	<tbody>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Disposable adhesive 
			surface silver/silver chloride electrodes</td>
			<td style="width:309px;">Spes 
			Medica S.r.l., Genova, Italy</td>
			<td style="width:119px;"></td>
			<td style="width:223px;">Self adhesive hydrogel electrode</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Abralyt HiCl</td>
			<td style="width:309px;">Easycap GmbH</td>
			<td style="width:119px;"></td>
			<td>Abrasive cream</td>
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			<td height="42" style="height:42px;width:216px;">Body Double fast</td>
			<td style="width:309px;">Smooth On Inc.</td>
			<td style="width:119px;"></td>
			<td>Skin adhesive silicone</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Telemetry system</td>
			<td style="width:309px;"></td>
			<td style="width:119px;"></td>
			<td>Internal development</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Picolog 1216</td>
			<td style="width:309px;">Pico Technology</td>
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			<td>AD converter</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Laptop</td>
			<td style="width:309px;">Panasonic</td>
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			<td>Rugged laptop</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Receiver</td>
			<td style="width:309px;"></td>
			<td style="width:119px;"></td>
			<td>Internal development</td>
		</tr>
	</tbody>
</table>

noninvasive eeg recordings from freely moving piglets

Il metodo permette la registrazione di alta qualità elettroencefalogrammi (EEG) di muoversi liberamente suinetti direttamente nel porcile. Usiamo un sistema telemetrico elettroencefalografia un canale in combinazione con gli elettrodi autoadesivi standard idrogel. I suinetti sono calmati giù senza l'uso di sedativi. Dopo il loro rilascio in porcile, i suinetti si comportano normalmente — bevono e dormire nello stesso ciclo come i loro fratelli. Loro fasi di sonno sono utilizzati per le registrazioni di EEG.

Siamo stati in grado di registrare tipico pattern EEG associate con non - REM del sonno, come mandrino scoppi o spazzole di delta, di muoversi liberamente suinetti (Figura 1 e Figura 2). Siamo stati per lo più interessati a modelli di rappresentanza durante non - REM del sonno, ma sono state anche fasi di sonno REM-come12 con una bassissima ampiezza registrata (Figura 3). La ...

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

Noninvasive EEG Recordings from Freely Moving Piglets

Qui, presentiamo un protocollo per registrare telemetrici elettroencefalogrammi (EEG) di muoversi liberamente suinetti direttamente nel porcile senza l'uso di sedativi, rendendo possibile registrare tipico pattern EEG durante non - REM del sonno, come mandrino scoppia.

Registrazioni di EEG non invadente da muoversi liberamente suinetti

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel ...

noninvasive-eeg-recordings-from-freely-moving-piglets

Research

JoVE Journal

Neuroscience

7.2K Views.  Free University Berlin. Qui, presentiamo un protocollo per registrare telemetrici elettroencefalogrammi (EEG) di muoversi liberamente suinetti direttamente nel porcile senza l'uso di sedativi, rendendo possibile registrare tipico pattern EEG durante non - REM del sonno, come mandrino scoppia.

Video: Noninvasive EEG Recordings from Freely Moving Piglets

High-density EEG Recordings of the Freely Moving Mice using Polyimide-based Microelectrode

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a noninvasive brain monitoring tool in cognitive neuroscience as well as a diagnostic tool for epilepsy and sleep disorders in neurology. However, the underlying mechanism of EEG rhythm generation is still under the veil. Recently introduced polyimide-based microelectrode (PBM-array) for high resolution mouse EEG1 is one of the trials to answer the neurophysiological questions on EEG signals based on a rich genetic resource that the mouse model contains for the analysis of complex EEG generation process. This application of nanofabricated PBM-array to mouse skull is an efficient tool for collecting large-scale brain activity of transgenic mice and accommodates to identify the neural correlates to certain EEG rhythms in conjunction with behavior. However its ultra-thin thickness and bifurcated structure cause a trouble in handling and implantation of PBM-array. In the presented video, the preparation and surgery steps for the implantation of PBM-array on a mouse skull are described step by step. Handling and surgery tips to help researchers succeed in implantation are also provided.

In this article, we described the surgery procedure and handling tips for implantation of ultra-thin polyimide-based microelectrode array (PBM-array) on the mouse skull for acquisition of high-density encephalography (EEG) in a mouse model.

Le registrazioni EEG ad alta densità dei Topi muoversi liberamente utilizzando Polyimide a base di microelettrodi

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

Ricerca

Neuroscienze

Patch Clamp Recordings from Embryonic Zebrafish Mauthner Cells

Mauthner cells (M-cells) are large reticulospinal neurons located in the hindbrain of teleost fish. They are key neurons involved in a characteristic behavior known as the C-start or escape response that occurs when the organism perceives a threat. The M-cell has been extensively studied in adult goldfish where it has been shown to receive a wide range of excitatory, inhibitory and neuromodulatory signals1. We have been examining M-cell activity in embryonic zebrafish in order to study aspects of synaptic development in a vertebrate preparation. In the late 1990s Ali and colleagues developed a preparation for patch clamp recording from M-cells in zebrafish embryos, in which the CNS was largely intact2,3,4. The objective at that time was to record synaptic activity from hindbrain neurons, spinal cord neurons and trunk skeletal muscle while maintaining functional synaptic connections within an intact brain-spinal cord preparation. This preparation is still used in our laboratory today. To examine the mechanisms underlying developmental synaptic plasticity, we record excitatory (AMPA and NMDA-mediated)5,6 and inhibitory (GABA and glycine) synaptic currents from developing M-cells. Importantly, this unique preparation allows us to return to the same cell (M-cell) from preparation to preparation to carefully examine synaptic plasticity and neuro-development in an embryonic organism. The benefits provided by this preparation include 1) intact, functional synaptic connections onto the M-cell, 2) relatively inexpensive preparations, 3) a large supply of readily available embryos 4) the ability to return to the same cell type (i.e. M-cell) in every preparation, so that synaptic development at the level of an individual cell can be examined from fish to fish, and 5) imaging of whole preparations due to the transparent nature of the embryos.

We have developed an intact brain-spinal cord preparation to record and monitor electrical activity via patch clamp recording from the Mauthner neurons and other reticulospinal cells in zebrafish embryos. Thus, we are able to record excitatory and inhibitory synaptic currents, voltage-gated channel activity and action potentials from key neurons in a developing embryo.

Bloccare Registrazioni patch da cellule embrionali Zebrafish Mauthner

Mauthner cells (M-cells) are large  reticulospinal neurons located in the hindbrain of teleost fish. They are key  neurons involved in a characteristic ...

Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed single-neuron activity while animals are acquiring an operantly conditioned response, or when that response is extinguished. But even in these cases, observation periods usually encompass only a single stage of learning, i.e. acquisition or extinction, but not both (exceptions include protocols employing reversal learning; see Bingman&#160;et al.1 for an example). However, acquisition and extinction entail different learning mechanisms and are therefore expected to be accompanied by different types and/or loci of neural plasticity.
Accordingly, we developed a behavioral paradigm which institutes three stages of learning in a single behavioral session and which is well suited for the simultaneous recording of single neurons&#39; action potentials. Animals are trained on a single-interval forced choice task which requires mapping each of two possible choice responses to the presentation of different novel visual stimuli (acquisition). After having reached a predefined performance criterion, one of the two choice responses is no longer reinforced (extinction). Following a certain decrement in performance level, correct responses are reinforced again (reacquisition). By using a new set of stimuli in every session, animals can undergo the acquisition-extinction-reacquisition process repeatedly. Because all three stages of learning occur in a single behavioral session, the paradigm is ideal for the simultaneous observation of the spiking output of multiple single neurons. We use pigeons as model systems, but the task can easily be adapted to any other species capable of conditioned discrimination learning.

Learning new stimulus-response associations engages a wide range of neural processes which are ultimately reflected in changing spike output of individual neurons. Here we describe a behavioral protocol allowing for the continuous registration of single-neuron activity while animals acquire, extinguish, and reacquire a conditioned response within a single experimental session.

Registrazione di potenziali d&#39;azione singoli neuroni &#39;di muoversi liberamente Piccioni attraverso tre fasi di apprendimento

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed ...

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees (Bombus terrestris)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from flowering plants. Only a few studies have described the gustatory responses of bees toward toxins in food, and these experiments have mainly used the proboscis extension response on restrained honey bees. Here, a new behavioral assay is presented for measuring the feeding responses of freely-moving, individual worker bumble bees to nutrients and toxins. This assay measures the amount of solution ingested by each bumble bee and identifies how tastants in food influence the microstructure of the feeding behavior.
The solutions are presented in a microcapillary tube to individual bumble bees that have been previously starved for 2-4 hr. The behavior is captured on digital video. The fine structure of the feeding behavior is analyzed by continuously scoring the position of the proboscis (mouthparts) from video recordings using event logging software. The position of the proboscis is defined by three different behavioral categories: (1) proboscis is extended and in contact with the solution, (2) proboscis is extended but not in contact with the solution and (3) proboscis is stowed under the head. Furthermore the speed of the proboscis retracting away from the solution is also estimated.
In the present assay the volume of solution consumed, the number of feeding bouts, the duration of the feeding bouts and the speed of the proboscis retraction after the first contact is used to evaluate the phagostimulatory or the deterrent activity of the compounds tested.
This new taste assay will allow researchers to measure how compounds found in nectar influence the feeding behavior of bees and will also be useful to pollination biologists, toxicologists and neuroethologists studying the bumble bee&#39;s taste system.

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees Bombus terrestris

A novel behavioral assay is described for investigating the short term gustatory responses of the mouthparts of freely-moving bumble bees (Bombus terrestris) toward nutrients and toxins in solution.

A Novel comportamentale test per Indagare Risposte gustativa individuale, liberamente rigiro bombi (<em&gt; Bombus terrestris</em&gt;)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from ...

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such as epilepsy and neurodegenerative disorders. However, it is technically difficult to obtain EEG recordings in neonates as it requires specialized handling and great care. Here, we present a novel method to record EEG in neonatal rat pups (P8-P15). We designed a simple and reliable electrode using computer pin loci; it can be easily implanted into the skull of a rat pup to record high-quality EEG signals in the normal and epileptic brain. Pups were given an intraperitoneal (i.p.) injection of the neurotoxin kainic acid (KA) to induce epileptic seizures. The surgical implantation performed in this procedure is less expensive than other EEG procedures for neonates. This method allows one to record high-quality and stable EEG signals for more than 1 week. Furthermore, this procedure can also be applied to adult rats and mice to study epilepsy or other neurological disorders.

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

Registrazione di EEG nei ratti neonatali liberamente in movimento utilizzando un metodo novello

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

Visual Evoked Potential Recordings in Mice Using a Dry Non-invasive Multi-channel Scalp EEG Sensor

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable because of the plunger-spring-barrel structural facet and mechanical strengths resulting from metal materials. The whole process for acquiring the VEP responses in vivo from a mouse consists of four steps: (1) sensor assembly, (2) animal preparation, (3) VEP measurement, and (4) signal processing. This paper presents representative measurements of VEP responses from multiple mice with a submicro-voltage signal resolution and sub-hundred millisecond temporal resolution. Although the proposed method is safer and more convenient compared to other previously reported animal EEG acquiring methods, there are remaining issues including how to enhance the signal-to-noise ratio and how to apply this technique with freely moving animals. The proposed method utilizes easily available resources and shows a repetitive VEP response with a satisfactory signal quality. Therefore, this method could be utilized for longitudinal experimental studies and reliable translational research exploiting non-invasive paradigms.

We designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable. This paper describes the whole process from manufacturing the proposed EEG electrode to signal processing of visual evoked potential (VEP) signals measured on a mouse scalp using a dry non-invasive multi-channel EEG sensor.

Registrazioni di potenziali evocate visive nei topi utilizzando un multicanale Non invasiva a secco del cuoio capelluto EEG sensore

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

Registrazioni simultanee di potenziali di campo locale corticale, elettrocardiogramma, elettromiogramma e respirazione ritmo da un ratto liberamente commovente

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain interstitial space with the behavior and/or with the specific outcome of a pathology (e.g., seizures for epilepsy). When studying epilepsy, the microdialysis technique is often combined with short-term or even long-term video-electroencephalography (EEG) monitoring to assess spontaneous seizure frequency, severity, progression and clustering. The combined microdialysis-EEG is based on the use of several methods and instruments. Here, we performed in vivo microdialysis and continuous video-EEG recording to monitor glutamate and aspartate outflow over time, in different phases of the natural history of epilepsy in a rat model. This combined approach allows the pairing of changes in the neurotransmitter release with specific stages of the disease development and progression. The amino acid concentration in the dialysate was determined by liquid chromatography. Here, we describe the methods and outline the principal precautionary measures one should take during in vivo microdialysis-EEG, with particular attention to the stereotaxic surgery, basal and high potassium stimulation during microdialysis, depth electrode EEG recording and high-performance liquid chromatography analysis of aspartate and glutamate in the dialysate. This approach may be adapted to test a variety of drug or disease induced changes of the physiological concentrations of aspartate and glutamate in the brain. Depending on the availability of an appropriate analytical assay, it may be further used to test different soluble molecules when employing EEG recording at the same time.

Here, we describe a method for in vivo microdialysis to analyze aspartate and glutamate release in the ventral hippocampus of epileptic and non-epileptic rats, in combination with EEG recordings. Extracellular concentrations of aspartate and glutamate may be correlated with the different phases of the disease.

Microdialisi degli aminoacidi eccitanti durante le registrazioni di EEG in liberi di muoversi

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain ...

Long-term Sensory Conflict in Freely Behaving Mice

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning in mice. By permanently wearing a device fixed on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages. Therefore, this protocol readily enables the study of the visual system and multisensory interactions over an extended timeframe that would not be accessible otherwise. In addition to lowering the experimental costs of long-term sensory learning in naturally behaving mice, this approach accommodates the combination of in vivo and in vitro experiments. In the reported example, video-oculography is performed to quantify the vestibulo-ocular reflex (VOR) and optokinetic reflex (OKR) before and after learning. Mice exposed to this long-term sensory conflict between visual and vestibular inputs presented a strong VOR gain decrease but exhibited few OKR changes. Detailed steps of device assembly, animal care, and reflex measurements are hereby reported.

The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning. By permanently wearing a fixed device on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages.

A lungo termine conflitto sensoriale a comportarsi liberamente topi

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for ...

Hybrid Microdrive System with Recoverable Opto-Silicon Probe and Tetrode for Dual-Site High Density Recording in Freely Moving Mice

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, conventional microdrive designs often only allow use of one type of electrode to record from single or multiple regions, limiting the yield of single-unit or depth profile recordings. It also often limits the ability to combine electrode recordings with optogenetic tools to target pathway and/or cell type specific activity. Presented here is a hybrid microdrive array for freely moving mice to optimize yield and a description of its fabrication and reuse of the microdrive array. The current design employs nine tetrodes and one opto-silicon probe implanted in two different brain areas simultaneously in freely moving mice. The tetrodes and the opto-silicon probe are independently adjustable along the dorsoventral axis in the brain to maximize the yield of unit and oscillatory activities. This microdrive array also incorporates a set-up for light, mediating optogenetic manipulation to investigate the regional- or cell type-specific responses and functions of long-range neural circuits. In addition, the opto-silicon probe can be safely recovered and reused after each experiment. Because the microdrive array consists of 3D-printed parts, the design of microdrives can be easily modified to accommodate various settings. First described is the design of the microdrive array and how to attach the optical fiber to a silicon probe for optogenetics experiments, followed by fabrication of the tetrode bundle and implantation of the array into a mouse brain. The recording of local field potentials and unit spiking combined with optogenetic stimulation also demonstrate feasibility of the microdrive array system in freely moving mice.

This protocol describes the construction of a hybrid microdrive array that allows implantation of nine independently adjustable tetrodes and one adjustable opto-silicon probe in two brain regions in freely moving mice. Also demonstrated is a method for safely recovering and reusing the opto-silicon probe for multiple purposes.

Sistema microdrive ibrido con sonda Opto-Silicon recuperabile e tetrode per la registrazione ad alta densità a doppio sito in mouse in movimento libero

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