Diese Arbeit wurde vom National Institute on Taubheit und andere Communication Disorders Grants R21DC013915 und R01DC012502, die Natur- und Ingenieurwissenschaften Research Council of Canada und der Europäische Forschungsrat des Siebten Rahmenprogramms der Europäischen Gemeinschaft (FP7 / 2007-2013 Finanzhilfevereinbarung Nr 339.152 unterstützt. ).

Die aktuelle experimentelle Protokoll folgt den Richtlinien für ethisches Verhalten nach der Yale University Menschenuntersuchungsausschuss. 1. Electroenchephalopgaphy (EEG) Herstellung <ol><li> Messen Sie die Kopfgröße, um die entsprechende EEG-Haube zu bestimmen. </li><li> Identifizieren Sie die Lage der Scheitelpunkt von der Suche nach der Mitte zwischen Nasion und Inion mit einem Maßband. </li><li> Legen Sie die EEG-Haube auf dem Kopf mit dem vorher festgelegten Scheitelpunkt als Cz. Untersuchen Cz wieder, nachdem Sie den Deckel, indem Sie ein Maßband wie in 1.2 getan. Man beachte, daß das EEG Kappe ist mit Elektrodenhalterungen ausgestattet ist, und die Platzierung der Elektroden 64 (oder Halter) auf einem modifizierten 10-20-System mit im Voraus festgelegten Koordinatensystem mit Cz 11 basiert. Hinweis: Diese repräsentative Anwendung eine 64-Elektroden-Konfiguration, um die Kopfhaut Verteilung Änderungen und zur Quellenanalyse. Für einfachere Anwendungen (ereignisbezogene möglichen Veränderungen in der Amplitude und Latenz) using weniger Elektroden sind möglich. Es gibt zwei zusätzliche Elektroden zum Boden in der hier verwendeten EEG-System. Diese Elektrodenhalter werden auch in der Kappe enthalten. </li><li> Bewerben Elektroden-Gel in den Elektrodenhalter mit einer Einwegspritze. </li><li> Befestigen EEG-Elektroden (einschließlich Masseelektroden) in den Elektroden Halter passend zu den Markierungen der Elektroden und der Elektrodenhalter auf der Elektrodenkappe. </li><li> Reinigen Sie die Oberfläche der Haut mit Alkohol-Pads. Anmerkung: Für Elektroden zum Erfassen Augenbewegung (elektro Okulographie) sind die Haut Stellen oberhalb und unterhalb des rechten Auges (vertikale Augenbewegung) und lateral von der äußeren Augenwinkel der beiden Augen (horizontale Augenbewegung); für somatosensorische Stimulation die Haut seitlich der Mundwinkel wird gereinigt. </li><li> Füllen Sie die vier elektro Okulographie Elektroden mit der Elektroden-Gel und sichern Sie die Elektroden mit doppelseitigem Klebeband auf die Standorte in 1.6 festgestellt. </li><li> Sichern Sie alle Elektrodenkabel mit einem Klettverschluss. Wenn required, kleben Sie die Kabel an Körper Teilnehmers oder die anderen Standorte, die keine zusätzliche elektrische oder mechanische Rauschen einführen. </li><li> Positionieren Sie den Teilnehmer vor dem Monitor und den Roboter zur somatosensorischen Stimulation. Sichern Sie alle Elektrodenkabel wieder wie in 1.8. </li><li> Schließen Sie das EEG und elektro Okulographie Elektroden (einschließlich der Masseelektroden) in die entsprechenden Verbindungsstücke (passenden Etikett und connecter Form) am Verstärker Feld des EEG-Systems. </li><li> Überprüfen Sie, dass die EEG-Signale artefaktfreie und dass der Offset-Wert in einem akzeptablen Bereich (&lt;50 &amp; mgr; V oder kleiner) ist sind. Wenn verrauschte Signale oder große Offsets, die normalerweise anzeigt hochohmig sind gefunden werden, diese richtige Elektrodensignale durch Hinzufügen zusätzlicher EEG-Gel und / oder Neupositionierung Haare, die direkt unter der Elektrode. </li><li> Legen Sie die EEG-kompatible Kopfhörer und bestätigen, dass der Schallpegel in einem angenehmen Bereich basierend auf Thema Bericht. </li></ol><pclass = &quot;jove_title&quot;&gt; 2. Somatosensorischen Stimulation Hinweis: Die aktuelle Protokoll gilt Gesichtshaut Dehnungs zum Zwecke der somatosensorische Stimulation. Der Versuchsaufbau mit der EEG-System ist in Figur 1 dargestellt ist. Die Einzelheiten der somatosensorischen Stimulationsvorrichtung sind in den früheren Studien 1,7,12-14 beschrieben. Kurz gesagt, werden zwei kleine Plastiklaschen (2 cm breite und 3 cm Höhe) mit doppelseitigem Klebeband auf die Gesichtshaut befestigt. Die Registerkarten werden in der Robotereinrichtung verbunden mit String. Der Roboter generiert systematische Hautdehnungsbelastungen nach experimentellen Designs. Das Setup-Protokoll für die ERP-Aufnahme ist wie folgt: <ol><li> Legen Sie den Kopf des Teilnehmers in der Kopfstütze, um die Kopfbewegung während der Stimulation zu minimieren. Entfernen Sie vorsichtig die Elektrodenkabel zwischen Kopf und Kopfstütze des Teilnehmers. </li><li> Bitten Sie die Teilnehmer, um den Sicherheitsschalter für den Roboter zu halten. </li><li> Befestigen Sie den Kunststofflaschendie Zielhautstelle mit doppelseitigem Klebeband für somatosensorische Stimulation. Für die repräsentative Ergebnisse 12,13, bei dem das Ziel ist die Haut lateral der Mundwinkel, mit der Mitte der Laschen an der Schneckenachse, wenige mm seitlich der Mundwinkel in der Mitte der Laschen etwa in der gleichen Höhe der Mundwinkel. </li><li> Einstellen der Konfiguration der Schnur, Bindfaden Stützen und des Roboters, um EEG-Elektroden und Leitungen zu vermeiden. </li><li> Geben Sie einige Gesichtshaut Strecken (ein Zyklus Sinuskurve bei 3 Hz mit einer maximalen Kraft von 4 N), um Artefakte zu überprüfen aufgrund der Stimulation (in der Regel als relativ große Amplitude und niedrigere Frequenz beobachtet, verglichen mit der elektrophysiologischen Antwort). Wenn Artefakte in den EEG-Signale beobachtet, gehen Sie zurück auf 2,4. </li></ol> 3. ERP-Recording <ol><li> Erläutern Sie die experimentelle Aufgabe, das Thema und bieten Praxis-Studien (ein Block = 10 Studien oder weniger), um zu bestätigen, wenn das Thema understands die Aufgabe klar. Hinweis: Die experimentelle Aufgabe und Reizdarbietung für ERP Aufnahme sind vorprogrammiert in Software für die Reizdarbietung. <ol><li> In der repräsentativen Test mit kombinierten somatosensorischen und auditive Stimulation 12, gelten die somatosensorische Stimulation mit Hautverformung in die Mundwinkel zugeordnet ist, um die Haut lateral. Das Muster der Strecke ist ein ein Zyklus Sinuskurve (3 Hz) mit einer maximalen Kraft von 4 N. Eine einzelne synthetisierte Sprachäußerung, die auf halber Strecke befindet sich in einem 10-Schritt-Sound Kontinuum zwischen &quot;Kopf&quot; und &quot;war&quot; ist für Gehörstimulation eingesetzt. </li><li> Präsentieren sowohl Stimulationen einzeln oder in Kombination. In der kombinierten Stimulation, Test drei Einsetzen Zeiten (90 ms Blei und Verzögerung, und gleichzeitiges in somatosensorischen und auditive Onsets: siehe 3A). </li><li> Randomize die Präsentation von fünf Stimulierungen (somatosensorische allein, auditive allein und drei kombinierten:. Blei, simult und lag). Variieren Sie die inter-trial-Intervall zwischen 1000 und 2000 ms, um die Vorfreude und Gewöhnung zu vermeiden. Die experimentelle Aufgabe ist es, durch Drücken einer Taste auf einer Tastatur zu identifizieren, ob die vorgelegten Sprachklang, die der Ton, der akustisch in der Mitte zwischen &quot;Kopf&quot; ist und &quot;musste&quot;, war &quot;Kopf&quot;. In der somatosensorischen allein Zustand, in dem es keine Hörstimulation, die Teilnehmer sind angewiesen, nicht &quot;Kopf&quot; zu beantworten. </li><li> Rekordteilnehmer Urteilen und die Reaktionszeit vom Beginn des Stimulus an den Tastendruck mit der Software für die Reizdarbietung. Bitten Sie die Teilnehmer, um einen Befestigungspunkt auf dem Bildschirm, um Artefakte aufgrund von Augenbewegungen zu reduzieren bestaunen. </li><li> Entfernen Sie den Befestigungspunkt alle 10 Stimulationen für einen Kurzurlaub. (Siehe auch die anderen Beispiel der Aufgabe und Reizdarbietung 12,13) </li></ol></li><li> Starten Sie die Software für ERP Aufnahme bei 512 HzProbenahme, die auch zeichnet die Anfangszeit der Stimulation in der Timeline von ERP-Daten. Beachten Sie, dass die Zeitstempel der Stimulation, die auch die Information über die Art der Stimulation, werden für jeden Stimulus von der Software für die Stimuluspräsentation gesendet. Die beiden Programme (ERP Aufnahme und für die Reizdarbietung) auf zwei getrennten PCs, die über einen Parallelanschluss verbunden sind aktiv. </li><li> Legen Sie die Software für den somatosensorischen Stimulation auf den Trigger-Wartemodus und starten Sie dann Reizdarbietung durch die Aktivierung der Software für die Reizdarbietung. Beachten Sie, dass die Software für den somatosensorischen Stimulation auch auf einem separaten PC laufen von den beiden anderen PCs. Nehmen Sie 100 ERPs pro Bedingung. Hinweis: Ein Triggersignal für den somatosensorischen Stimulation über eine analoge Eingabevorrichtung, die an ein digitales Ausgabegerät im PC für sensorische Stimulation verbunden wird empfangen. Einzel somatosensorischen Stimulation pro Trigger produziert. &lt;/ li&gt; </ol>

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The authors have nothing to disclose.

Die hier ausgewiesenen Studien belegen, die präzise gesteuert somatosensorischen Stimulation, die durch die Gesichtshaut Verformung erzeugt wird, induziert kortikalen ERPs. Kutanen Afferenzen werden als eine reiche Quelle von kinästhetische Informations 3,4 in der menschlichen Bewegung des Körpers 5,6 und Sprachbewegung 7,8,21 bekannt. Dehnen der Gesichtshaut in der Weise, dass die tatsächliche Bewegungsrichtung während des Sprechens reflektiert induziert eine kinästhetische Sinne ...

Sprachproduktion hängt sowohl von auditiven und somatosensorischen Informationen. Die auditiven und somatosensorischen Feedback treten in Kombination von den ersten Laute von einem Säugling produziert und beide sind in der Rede motorischen Lernens beteiligt. Jüngste Ergebnisse deuten darauf hin, dass somatosensorische Prozesse tragen zur Wahrnehmung sowie Produktion. So klingt die Identifikation der Sprache verändert, wenn ein Roboter-Vorrichtung erstreckt sich die Gesichtshaut als Teilnehmer hören auf akustische Reize 1. Air Puffs an der Wange, die mit auditiven Sprachreizen überein verändern Wahrnehmungsurteile Teilnehmer 2. Diese somatosensorischen Effekte beinhalten die Aktivierung von kutanen Mechanorezeptoren in Reaktion auf Hautverformung. Die Haut wird auf verschiedene Weisen während der Bewegung verformt und kutane Mechanoreceptoren führen bekanntlich zu kinästhetische Sinne 3,4 beizutragen. Der kinästhetische Rolle der kutanen Mechanorezeptoren ist demondurch die jüngsten Feststellungen 5-7, dass die Bewegung bedingte Hautstämme werden in geeigneter Weise, wie Beugung oder Streckung Bewegung je nach dem Muster der Haut Stretch 6 wahrgenommen strated. Im Laufe der Sprechmotorik Ausbildung, die die Wiederholung von bestimmten Sprachäußerung mit begleitender Gesichtshaut Dehnungs Rede ist, ändern Artikulationsmuster adaptiv 7. Diese Studien zeigen, dass die Modulation der Haut Stretch während der Aktion ist ein Verfahren zur Bewertung des Beitrags der kutanen Afferenzen auf die kinästhetische Funktion des sensomotorischen Systems. Der kinästhetische Funktion der orofazialen kutanen Mechanorezeptoren wurde vor allem mit psychophysiologischen Methoden 7,8 und Mikroelektroden-Umkodierung von sensorischen Nerven 9,10 sucht. Hier konzentriert sich das aktuelle Protokoll auf der Kombination von orofazialen somatosensorischen Stimulation mit Gesichtshaut Verformung und ereignisbezogenen Potenzial (ERP) Aufnahme verbunden. Thist Vorgehensweise hat genaue experimentelle Kontrolle über die Richtung und den Zeitpunkt der Gesichtshaut Verformung mit Hilfe eines computergesteuerten Robotervorrichtung. Dies erlaubt uns, spezifische Hypothesen über die somatosensorischen Beitrag zur Sprachproduktion und Wahrnehmung durch selektive und präzise Verformungsgesichtshaut in einer Vielzahl von Orientierungen sowohl während der Rede motorischen Lernens zu testen und direkt bei der Sprachproduktion und Wahrnehmung. ERP Aufzeichnung werden zur nichtinvasiven Auswertung der zeitlichen Muster und Zeitpunkt der Einfluß der somatosensorischen Stimulation auf orofacial Verhaltensweisen. Das aktuelle Protokoll kann dann die neuronalen Korrelate der kinästhetische Funktion auszuwerten und zu bewerten den Beitrag des somatosensorischen Systems sowohl für Sprachverarbeitung, Sprachproduktion und Sprachwahrnehmung. Um die Nützlichkeit der Anwendung von Hautdehnungs Stimulation ERP Aufnahme zeigen, konzentriert sich die folgende Protokoll auf der Wechselwirkung von somatosensorischen und auditive Eingabe in Rede perception. Die Ergebnisse zeigen eine potenzielle Methode zur somatosensorischen-auditive Interaktion in der Rede zu bewerten.

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somatosensory event-related potentials from orofacial skin stretch stimulation

Cortical processing associated with orofacial somatosensory function in speech has received limited experimental attention due to the difficulty of providing precise and controlled stimulation. This article introduces a technique for recording somatosensory event-related potentials (ERP) that uses a novel mechanical stimulation method involving skin deformation using a robotic device. Controlled deformation of the facial skin is used to modulate kinesthetic inputs through excitation of cutaneous mechanoreceptors. By combining somatosensory stimulation with electroencephalographic recording, somatosensory evoked responses can be successfully measured at the level of the cortex. Somatosensory stimulation can be combined with the stimulation of other sensory modalities to assess multisensory interactions. For speech, orofacial stimulation is combined with speech sound stimulation to assess the contribution of multi-sensory processing including the effects of timing differences. The ability to precisely control orofacial somatosensory stimulation during speech perception and speech production with ERP recording is an important tool that provides new insight into the neural organization and neural representations for speech.

Dieser Abschnitt stellt repräsentative Ereignis-korrelierte Potenziale in Reaktion auf somatosensorische Stimulation von Gesichtshaut Verformung resultieren. Der experimentelle Aufbau ist in Abbildung 1 dargestellt. Sinusoidal Stimulierung wurde auf die Gesichtshaut seitlich der Mundwinkel (siehe 3A als Referenz) aufgetragen. Einhundert Dehnungsversuche wurden für jeden Teilnehmer mit 12 Teilnehmern in insgesamt getestet aufgezeichnet....

Watch this Scientific Journal Video about Somatosensory Event-related Potentials from Orofacial Skin Stretch Stimulation at JoVE.com

Somatosensory Event-related Potentials from Orofacial Skin Stretch Stimulation

This paper introduces a method for obtaining somatosensory event-related potentials following orofacial skin stretch stimulation. The current method can be used to evaluate the contribution of somatosensory afferents to both speech production and speech perception.

Somatosensorischen Ereignis-korrelierte Potenziale von Orofaziale Haut Stretch Stimulation

Cortical processing associated with orofacial somatosensory function in speech has received limited experimental attention due to the difficulty of ...

somatosensory-event-related-potentials-from-orofacial-skin-stretch

Research

JoVE Journal

Neuroscience

13.1K Aufrufe.  Haskins Laboratories. This paper introduces a method for obtaining somatosensory event-related potentials following orofacial skin stretch stimulation. The current method can be used to evaluate the contribution of somatosensory afferents to both speech production and speech perception.

Serie: Somatosensory Event-related Potentials from Orofacial Skin Stretch Stimulation

High Density Event-related Potential  Data Acquisition in Cognitive Neuroscience

Functional magnetic resonance imaging (fMRI) is currently the standard method of evaluating brain function in the field of Cognitive Neuroscience, in part because fMRI data acquisition and analysis techniques are readily available. Because fMRI has excellent spatial resolution but poor temporal resolution, this method can only be used to identify the spatial location of brain activity associated with a given cognitive process (and reveals virtually nothing about the time course of brain activity). By contrast, event-related potential (ERP) recording, a method that is used much less frequently than fMRI, has excellent temporal resolution and thus can track rapid temporal modulations in neural activity. Unfortunately, ERPs are under utilized in Cognitive Neuroscience because data acquisition techniques are not readily available and low density ERP recording has poor spatial resolution. In an effort to foster the increased use of ERPs in Cognitive Neuroscience, the present article details key techniques involved in high density ERP data acquisition. Critically, high density ERPs offer the promise of excellent temporal resolution and good spatial resolution (or excellent spatial resolution if coupled with fMRI), which is necessary to capture the spatial-temporal dynamics of human brain function.

Event-related potential (ERP) recording is under utilized in Cognitive Neuroscience because data acquisition techniques are not readily available and this method often has poor spatial resolution. To foster the increased use of ERPs in Cognitive Neuroscience, the present article details key techniques involved in high density ERP data acquisition.

High Density Event-Related Potential Data Acquisition in Cognitive Neuroscience

Functional magnetic resonance imaging (fMRI) is currently the standard method of evaluating brain function in the field of Cognitive Neuroscience, in part ...

Forschung

Neurowissenschaften

Combining Transcranial Magnetic Stimulation and fMRI to Examine the Default Mode Network

The default mode network is a group of brain regions that are active when an individual is not focused on the outside world and the brain is at "wakeful rest."1,2,3 It is thought the default mode network corresponds to self-referential or "internal mentation".2,3
It has been hypothesized that, in humans, activity within the default mode network is correlated with certain pathologies (for instance, hyper-activation has been linked to schizophrenia 4,5,6 and autism spectrum disorders 7 whilst hypo-activation of the network has been linked to Alzheimer's and other neurodegenerative diseases 8). As such, noninvasive modulation of this network may represent a potential therapeutic intervention for a number of neurological and psychiatric pathologies linked to abnormal network activation. One possible tool to effect this modulation is Transcranial Magnetic Stimulation: a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability (either increasing or decreasing it) via the application of localized magnetic field pulses.9
In order to explore the default mode network's propensity towards and tolerance of modulation, we will be combining TMS (to the left inferior parietal lobe) with functional magnetic resonance imaging (fMRI). Through this article, we will examine the protocol and considerations necessary to successfully combine these two neuroscientific tools.

In this article, we examine the methodology and considerations relevant to the combination of TMS and fMRI to examine the effects of brain stimulation on the default network.

Die Kombination TMS und fMRT dem Standard-Modus Netzwerk Untersuchen

The default mode network is a group of brain regions that are active when an individual is not focused on the outside world and the brain is at "wakeful ...

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS display abnormalities in the expression of FMR1 - a protein required for typical, healthy neural development.3 Recent data has suggested that the loss of this protein can cause the cortex to be hyperexcitable thereby affecting overall patterns of neural plasticity.4,5
In addition, Fragile X shows a strong comorbidity with autism: in fact, 30% of children with FXS are diagnosed with autism, and 2 - 5% of autistic children suffer from FXS.6
Transcranial Magnetic Stimulation (a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability via the application of localized magnetic field pulses 7,8) represents a unique method of exploring plasticity and the manifestations of FXS within affected individuals. More specifically, Theta-Burst Stimulation (TBS), a specific stimulatory protocol shown to modulate cortical plasticity for a duration up to 30 minutes after stimulation cessation in healthy populations, has already proven an efficacious tool in the exploration of abnormal plasticity.9,10 
Recent studies have shown the effects of TBS last considerably longer in individuals on the autistic spectrum - up to 90 minutes.11 This extended effect-duration suggests an underlying abnormality in the brain's natural plasticity state in autistic individuals - similar to the hyperexcitability induced by Fragile X Syndrome.
In this experiment, utilizing single-pulse motor-evoked potentials (MEPs) as our benchmark, we will explore the effects of both intermittent and continuous TBS on cortical plasticity in individuals suffering from FXS and individuals on the Autistic Spectrum.

In this article, we examine the effects of Theta-Burst TMS stimulation on cortical plasticity in individuals suffering from Fragile X syndrome and individuals on the autistic spectrum.

TMS: Mit dem Theta-Burst-Protokoll zu Mechasnism der Plastizität in Individuen sich mit Fragile X-Syndrom und Autismus

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS ...

Electrode Positioning and Montage in Transcranial Direct Current Stimulation

Transcranial direct current stimulation (tDCS) is a technique that has been intensively investigated in the past decade as this method offers a non-invasive and safe alternative to change cortical excitability2. The effects of one session of tDCS can last for several minutes, and its effects depend on polarity of stimulation, such as that cathodal stimulation induces a decrease in cortical excitability, and anodal stimulation induces an increase in cortical excitability that may last beyond the duration of stimulation6. These effects have been explored in cognitive neuroscience and also clinically in a variety of neuropsychiatric disorders &#x2013; especially when applied over several consecutive sessions4. One area that has been attracting attention of neuroscientists and clinicians is the use of tDCS for modulation of pain-related neural networks3,5. Modulation of two main cortical areas in pain research has been explored: primary motor cortex and dorsolateral prefrontal cortex7. Due to the critical role of electrode montage, in this article, we show different alternatives for electrode placement for tDCS clinical trials on pain; discussing advantages and disadvantages of each method of stimulation.

Transcranial direct current stimulation (tDCS) is an established technique to modulate cortical excitability1,2. It has been used as an investigative tool in neuroscience due to its effects on cortical plasticity, easy operation, and safe profile. One area that tDCS has been showing encouraging results is pain alleviation 3-5.

Electrode Positionierung und Montage in Transkranielle Direct Current Stimulation

Transcranial direct current stimulation (tDCS) is a technique that has been intensively investigated in the past decade as this method offers a ...

Applications of EEG Neuroimaging Data: Event-related Potentials, Spectral Power, and Multiscale Entropy

When considering human neuroimaging data, an appreciation of signal variability represents a fundamental innovation in the way we think about brain signal. Typically, researchers represent the brain's response as the mean across repeated experimental trials and disregard signal fluctuations over time as "noise". However, it is becoming clear that brain signal variability conveys meaningful functional information about neural network dynamics. This article describes the novel method of multiscale entropy (MSE) for quantifying brain signal variability. MSE may be particularly informative of neural network dynamics because it shows timescale dependence and sensitivity to linear and nonlinear dynamics in the data.

Neuroimaging researchers typically consider the brain's response as the mean activity across repeated experimental trials and disregard signal variability over time as "noise". However, it is becoming clear that there is signal in that noise. This article describes the novel method of multiscale entropy for quantifying brain signal variability in the time domain.

Anwendungen von EEG Neuroimaging Daten: Ereignis-korrelierte Potenziale, Spectral Power und Multiscale Entropy

When considering human neuroimaging data, an appreciation of signal variability represents a fundamental innovation in the way we think about brain ...

Optogenetic Activation of Zebrafish Somatosensory Neurons using ChEF-tdTomato

Larval zebrafish are emerging as a model for describing the development and function of simple neural circuits. Due to their external fertilization, rapid development, and translucency, zebrafish are particularly well suited for optogenetic approaches to investigate neural circuit function. In this approach, light-sensitive ion channels are expressed in specific neurons, enabling the experimenter to activate or inhibit them at will and thus assess their contribution to specific behaviors. Applying these methods in larval zebrafish is conceptually simple but requires the optimization of technical details. Here we demonstrate a procedure for expressing a channelrhodopsin variant in larval zebrafish somatosensory neurons, photo-activating single cells, and recording the resulting behaviors. By introducing a few modifications to previously established methods, this approach could be used to elicit behavioral responses from single neurons activated up to at least 4 days post-fertilization (dpf). Specifically, we created a transgene using a somatosensory neuron enhancer, CREST3, to drive the expression of the tagged channelrhodopsin variant, ChEF-tdTomato. Injecting this transgene into 1-cell stage embryos results in mosaic expression in somatosensory neurons, which can be imaged with confocal microscopy. Illuminating identified cells in these animals with light from a 473 nm DPSS laser, guided through a fiber optic cable, elicits behaviors that can be recorded with a high-speed video camera and analyzed quantitatively. This technique could be adapted to study behaviors elicited by activating any zebrafish neuron. Combining this approach with genetic or pharmacological perturbations will be a powerful way to investigate circuit formation and function.

Optogenetic techniques have made it possible to study the contribution of specific neurons to behavior. We describe a method in larval zebrafish for activating single somatosensory neurons expressing a channelrhodopsin variant (ChEF) with a diode-pumped solid state (DPSS) laser and recording the elicited behaviors with a high-speed video camera.

Optogenetische Aktivierung der Zebrafisch Somatosensory Neuronen mit Chef-tdTomato

Larval  zebrafish are emerging as a model for describing the development and function  of simple neural circuits. Due to their  external fertilization, ...

Extracting Visual Evoked Potentials from EEG Data Recorded During fMRI-guided Transcranial Magnetic Stimulation

Transcranial Magnetic Stimulation (TMS) is an effective method for establishing a causal link between a cortical area and cognitive/neurophysiological effects. Specifically, by creating a transient interference with the normal activity of a target region and measuring changes in an electrophysiological signal, we can establish a causal link between the stimulated brain area or network and the electrophysiological signal that we record. If target brain areas are functionally defined with prior fMRI scan, TMS could be used to link the fMRI activations with evoked potentials recorded. However, conducting such experiments presents significant technical challenges given the high amplitude artifacts introduced into the EEG signal by the magnetic pulse, and the difficulty to successfully target areas that were functionally defined by fMRI. Here we describe a methodology for combining these three common tools: TMS, EEG, and fMRI. We explain how to guide the stimulator&#39;s coil to the desired target area using anatomical or functional MRI data, how to record EEG during concurrent TMS, how to design an ERP study suitable for EEG-TMS combination and how to extract reliable ERP from the recorded data. We will provide representative results from a previously published study, in which fMRI-guided TMS was used concurrently with EEG to show that the face-selective N1 and the body-selective N1 component of the ERP are associated with distinct neural networks in extrastriate cortex. This method allows us to combine the high spatial resolution of fMRI with the high temporal resolution of TMS and EEG and therefore obtain a comprehensive understanding of the neural basis of various cognitive processes.

This paper describes a method for collecting and analyzing electroencephalography (EEG) data during concurrent transcranial magnetic stimulation (TMS) guided by activations revealed with functional magnetic resonance imaging (fMRI). A method for TMS artifact removal and extraction of event related potentials is described as well as considerations in paradigm design and experimental setup.

Extrahieren von visuell evozierten Potentiale von EEG-Daten aufzeichnen Während fMRI-geführte Transkranielle Magnetstimulation

Transcranial Magnetic Stimulation (TMS) is an effective method for establishing a causal link between a cortical area and cognitive/neurophysiological ...

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.

Recording Single Neurons&#039; Action Potentials from Freely Moving Pigeons Across Three Stages of 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.

Aufnahme von Aktionspotentialen einzelner Neuronen &quot;sich frei zu bewegen Tauben auf drei Stages of Learning

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

Analysis of Developing Tooth Germ Innervation Using Microfluidic Co-culture Devices

Innervation plays a key role in the development, homeostasis and regeneration of organs and tissues. However, the mechanisms underlying these phenomena are not well understood yet. In particular, the role of innervation in tooth development and regeneration is neglected.
Several in vivo studies have provided important information about the patterns of innervation of dental tissues during development and repair processes of various animal models. However, most of these approaches are not optimal to highlight the molecular basis of the interactions between nerve fibres and target organs and tissues.
Co-cultures constitute a valuable method to investigate and manipulate the interactions between nerve fibres and teeth in a controlled and isolated environment. In the last decades, conventional co-cultures using the same culture medium have been performed for very short periods (e.g., two days) to investigate the attractive or repulsive effects of developing oral and dental tissues on sensory nerve fibres. However, extension of the culture period is required to investigate the effects of innervation on tooth morphogenesis and cytodifferentiation.
Microfluidics systems allow co-cultures of neurons and different cell types in their appropriate culture media. We have recently demonstrated that trigeminal ganglia (TG) and teeth are able to survive for a long period of time when co-cultured in microfluidic devices, and that they maintain in these conditions the same innervation pattern that they show in vivo.
On this basis, we describe how to isolate and co-culture developing trigeminal ganglia and tooth germs in a microfluidic co-culture system.This protocol describes a simple and flexible way to co-culture ganglia/nerves and target tissues and to study the roles of specific molecules on such interactions in a controlled and isolated environment.

Co-cultures represent a valuable method to study the interactions between nerves and target tissues and organs. Microfluidic systems allow co-culturing ganglia and whole developing organs or tissues in different culture media, thus representing a valuable tool for the in vitro study of the crosstalk between neurons and their targets.

Die Analyse der Entwicklung von Zahnkeim Innervation Mit Microfluidic Co-Kultur-Geräte

Innervation plays a key role in the development, homeostasis and regeneration of organs and tissues. However, the mechanisms underlying these phenomena ...

How to Find Effects of Stimulus Processing on Event Related Brain Potentials of Close Others when Hyperscanning Partners

The partners of each pair must be able to pass the McGill Friendship Questionnaire without communicating. Each partner is then seated in front of a screen in one of two adjacent rooms. These rooms are separated by a glass window through which participants communicate to maintain feelings of togetherness while being fitted with the EEG cap. After checking for adequate EEG signals, the glass is covered by a curtain to prevent visual communication. Then, partners must be silent but are instructed to try to feel in the presence of their partner during the entire experiment. Just before it starts, participants are told that each of them will be presented with one image at a time and that these images will occur at the same time for both of them on their own screen. They are also instructed that, for each trial, the simultaneous images will always be different. However, unbeknownst to them, trials are randomized: only half of them are consistent with this instruction and actually include two different images. These trials form the DSC, that is, the different-stimuli condition. The other half of the trials are inconsistent with the instruction. They include two identical images and form the ISC (identical-stimuli condition). After the experiment, participants are sorted into two groups: those who reported that they felt in the presence of their partner during the majority of the trials and those who reported they did not. The impact of the stimulus processing of the partner is found by subtracting the mean voltages of the ERPs of the ISC (inconsistent with the instructions) from the ERPs of the DSC (consistent with the instructions) in at least two time windows (TWs): firstly, in the 75 to 150 ms TW, where the absolute values of these subtractions are greater, especially at right frontal sites, in those who felt in the presence of their partner than in those who did not; secondly, in the LPP time window (i.e., from 650 to 950 ms post onset), where ERPs are significantly less positive in the DSC than in the ISC in those in whom the raw results of the early (75-150ms) subtractions are negative.

This protocol describes key steps involved in assessing the sensitivity of the brain of one person to the stimulus processing of a close other by selecting pairs of partners, recording their electroencephalogram (EEG) simultaneously and computing their event-related brain potentials (ERPs).

Wie Sie finden Effekte des Stimulus Verarbeitung auf Veranstaltung Verwandte Gehirn Potenziale der enge anderen wenn Hyperscanning Partner

The partners of each pair must be able to pass the McGill Friendship Questionnaire without communicating. Each partner is then seated in front of a screen ...