Name: simultaneous recording of electroretinography and visual evoked potentials in anesthetized rats
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Funding for this project was provided by the National Health and Medical Research Council (NHMRC) 1046203 (BVB, AJV) and Melbourne Neuroscience Institute Fellowship (CTN).

<ol>
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The authors have no disclosures relevant to this work.

Die ERG und VEP sind objektive Maßnahmen der visuellen Funktion von der Retina und Cortex sind. Der Vorteil der gleichzeitigen Aufnahme ist, dass eine umfassendere Sicht auf den gesamten Sehbahn wird gewährt. Im Konkreten könnte die komplementäre Informationen aus ihrer gleichzeitigen Beurteilung eine klarere Abgrenzung der Stelle der Verletzung in der Sehbahn bereitzustellen (beispielsweise zur Behandlung von Störungen mit überlappenden ERG noch deutliche VEP Manifestationen 18, wenn optische Neuropathie mit primären zerebrale Atrophie koexistieren kann 19, 20, oder wenn VEP Verlust kann durch Manifestation von Verletzungen an mehreren Standorten in Sehbahn 21,22) verwechselt werden. Durch die Messung gleichzeitig die ERG und VEP, ein Index der Verstärkung zwischen Retinal und kortikalen Reaktion kann auch abgeleitet werden. Dies kann ein nützliches Werkzeug zur Verfügung stellen subtile pathologischen Veränderungen zu erkennen. Das aktuelle Protokoll ermöglicht für ERG und VEP-Messung in Labor-Ratten häufig verwendet, kann aber ohne weiteres ad sein23-25 ​​Apted auf andere Säugetierarten. ERG und VEP Wellenformen von Nagetieren eine hinreichend sichere präklinische Surrogat für Reaktionen in menschlichen Augen beobachtet 26-28. Durch einen bestimmten Stimulus - Protokoll Gestaltung, sowohl ERG und VEP Reaktion kann während einer einzigen Aufzeichnungssitzung erhalten werden. Tabelle 1 zeigt eine Progression der Lichtstärke mit entsprechender Berücksichtigung der Erholungszeit zwischen den aufeinanderfolgenden blinkt. Dieses Protokoll bietet ein ausgewogenes Verhältnis zwischen der Notwendigkeit, Signal-zu-Rauscheigenschaften zu maximieren und die Aufnahmezeit im Anästhetikum Fenster durch eine einmalige Gabe von Ketamin zur Verfügung gestellt zu begrenzen: Xylazin. Daher kann diese Technik für eine objektive quantitative Maß der Sehfunktion zur Erforschung grundlegende Physiologie und Krankheiten nützlich sein. Eine umfassende Beurteilung des visuellen Systems kann durch gleichzeitige Beurteilung der bilateralen retinalen Antworten und visuell evozierten kortikalen Antworten erreicht werden. Jedoch kann jede Technik auch in Isolierung und monokular statt binokular durchgeführt werden, um das Verfahren zu vereinfachen. Das aktuelle Protokoll beschreibt scotopic ERG und VEP Signale gewählt, um den stab Weg gegeben, zu isolieren, dass Ratten, die eine Stange dominierten Netzhaut haben. Wenn das Licht angepasst Antworten von mehr Interesse für die Studie sind, ist es auch möglich, photopic ERG und VEP Signale, die von Pre-Anpassung an ein Hintergrundlicht zu leiten. Eine wesentliche Einschränkung dieser Technik ist die Notwendigkeit, das Verfahren unter narkotisiert Bedingungen durchzuführen stabile Platzierung der Elektroden zu ermöglichen. Dennoch bietet dieser Ansatz robustes Signal-zu-Rauscheigenschaften ermöglicht Erkennung von subtilen Behandlung ändert. Aufgrund der geringen Amplitude der STR und ihre Lichtempfindlichkeit Anpassung, müssen mehrere Schritte genau beobachtet werden erfolgreiche Aufnahme dieser Reaktion zu gewährleisten. Erstens muss eine ausreichende Dunkeladaptation umgesetzt werden, die beinhaltetüber Nacht Dunkeladaptation (≥ 8 h), Elektrodenplatzierung bei schwacher roter Beleuchtung (17,4 cd.m -2, λ max = 600 nm) und erneut Dunkeladaptation nach einem dim Test-Flash (10 min für - 0,52 log cd. sm -2). Weiterhin kann das Signal-zu-Rausch - Charakteristika des STR durch Mittelung über mehrere Signale verbessert werden (dh 20 - Signale) mit kurzen Inter-Stimulus - Intervallen gesammelt (dh 2 sec). Einer der Vorteile dieser umfassenden Bewertung beider Augen und cortices 3 Vergleich zur kontralateralen Aufnahme zu ermöglichen. Als solche sollten besondere Sorgfalt (dh gleiche Größe und förmigen Elektroden) in der Elektrodenherstellung genommen werden, um minimal inter Auge und inter kortikalen Variabilität zu gewährleisten. Angesichts der umfangreichen Nutzung beider ERG und VEP Techniken in vivo Maßnahmen der Sehbahn und seiner krankheitsbedingten Prozesse zur Verfügung zu stellen, wäre es sinnvoll sein , andere Weg spezifische p zusammenzutragenrotocols (beispielsweise EIN / AUS oder kegelUnterTyp - spezifische) und gleichzeitige Aufnahmen ERG / VEP zuführen mit unterschiedlichen Reizmodalitäten (beispielsweise Flimmern, Muster, Sägezahn) , um die Anwendung dieser Technik in klinischen Diagnosen zu erweitern. Ein weiterer logischer Schritt dieser Anwendung in der Zukunft wäre auch die ERG und VEP aufzeichnen gleichzeitig von bewussten 29, Tiere frei bewegten Anästhetikum Einflüsse 30 auf neuronale Physiologie zu vermeiden.

Messung der Elektroretinogramm (ERG) und visuell evozierte Potential (VEP) liefern nützliche quantitative Beurteilung der Integrität der Sehbahn. Die ERG misst die elektrischen Antworten der Netzhaut auf Licht Stimulation, während der VEP die entsprechende funktionale Integrität der Sehbahn von der Netzhaut zum primären visuellen Kortex nach dem gleichen Lichtereignis misst. Dieses Manuskript beschreibt ein Protokoll für die Erfassung und Analyse von ERG und VEP-Antworten in einem allgemein verwendeten Labormodell, der Ratte. Die ERG liefert einen Index der funktionellen Integrität einer Reihe von wichtigen Netzhautzellklassen durch die Netzhaut des elektrischen Brutto Reaktion auf einen Lichtblitz zu quantifizieren. Eine koordinierte Reihe von Ionenflüssen durch Licht initiierte Einsetzen und Offset erzeugen erkennbaren Änderungen in der Spannung, die gemessen werden können, außerhalb des Auges platziert Oberflächenelektroden verwendet. Die sich ergebende Wellenform stellt die Kombination einer seRies von gut definierten Komponenten, in Amplitude, Zeitsteuerung und Frequenz unterscheiden. Eine erhebliche Anzahl von Studien hat gezeigt, dass diese Komponenten sind relativ gut in vielen wirbel Retinae konservierten und dass die Komponenten voneinander getrennt werden können. Durch umsichtig den Reiz (Flash - Stimulus, Hintergrund, Interstimulus Intervall) Auswählen von Bedingungen und spezifischen Merkmale der zusammengesetzten Wellenform der Wahl zu analysieren kann man ein Maß für eine bestimmte Gruppe von Netzhautzellen von der Rückkehr 1,2 zuversichtlich sein. Diese Eigenschaften unterliegen die Brauchbarkeit und somit die weit verbreitete Anwendungen der ERG als nicht-invasive Messung der Netzhautfunktion. Diese Handschrift konzentriert sich auf die Methodik für die ERG die Messung und Analyse ihrer Funktionen Informationen über einige der wichtigsten Zellklassen in der Netzhaut zurück, nämlich Photorezeptoren (die PIII-Komponente), bipolare Zellen (die PII-Komponente) und retinale Ganglionzellen (die positive scotopic Schwellenreaktion oder pstr). <p class= &quot;Jove_content&quot;&gt; Das VEP stellt einen Assay der kortikalen Reaktion auf Licht; zuerst von der Netzhaut stamm und danach seriell über den Sehnerv, Sehbahn, Thalamus (Corpus geniculatum laterale, LGN) und optische Strahlung Bereich V1 des Kortex 3 mitgeteilt. Bei Nagetieren, die Mehrheit (90-95%) der Sehnervenfasern von jedem Auge decussate 4 und innervate kontralateralen Mittelhirns. Anders als bei der ERG, ist es noch nicht möglich , verschiedene Komponenten der VEP an spezifische Zellklassen zuzuordnen, 5 somit überall ändert sich entlang der Sehbahn die VEP - Wellenform beeinflussen könnten. Dennoch ist die VEP eine nützliche nicht-invasive Messung der Sehleistung und Sehbahn Integrität. Die VEP, wenn sie in Verbindung mit dem ERG verwendet wird , kann eine vollständigere Beurteilung der visuellen Systems bereitzustellen (dh Retina / Sehbahn). ERG und VEP-Aufnahmen können für sich allein oder in Kombination durchgeführt werden, abhängig von der AnwenKation. Die Methodik in diesem Papier beschrieben ermöglicht die gleichzeitige Beurteilung der Netzhaut und im kortikalen visuell evozierten Elektrophysiologie von beiden Augen und die beiden Hemisphären in narkotisierten Ratten. Dies ist ein nützlicher Weg zu umfassender Netzhautfunktion und die Upstream-Effekte zu bewerten, dass Veränderungen der Netzhautfunktion auf visuell evozierten kortikalen Funktion haben kann.

<table><tbody><tr><td>Alligator clip</td><td>generic brand</td><td>HM3022</td><td>Stainless steel 26 mm clip for connecting VEP screw electrodes to cables</td></tr><tr><td>Bioamplifier</td><td>ADInstruments</td><td>ML 135</td><td>For amplifying ERG and VEP signals</td></tr><tr><td>Carboxymethylcellulose sodium 1.0%</td><td>Allergan</td><td>CAS 0009000-11-7</td><td>Viscous fluid for improving signal quality of the active ERG electrode</td></tr><tr><td>Carprofen 0.5%</td><td>Pfizer Animal Health Group</td><td>CAS 53716-49-7</td><td>Proprietary name: Rimadyl injectable (50 mg/ml). For post-surgery analgesia, diluted to 0.5% (5 mg/ml) in normal saline</td></tr><tr><td>Chlorhexadine 0.5%</td><td>Orion Laboratories</td><td>27411, 80085</td><td>For disinfecting surgical instruments</td></tr><tr><td>Circulating water bath</td><td>Lauda-K&amp;ouml;nigshoffen</td><td>MGW Lauda</td><td>For maintaining body temperature of the anesthetized animal during surgery and electrophysiological recordings</td></tr><tr><td>Dental amalgam</td><td>DeguDent GmbH</td><td>64020024</td><td>For encasing the electrode-skull assembly to make it more robust</td></tr><tr><td>Dental burr</td><td>Storz Instruments, Bausch and Lomb</td><td>#E0824A</td><td>A miniature drill head of ~ 0.7 mm diameter for making a small hole in the skull over each hemisphere to implant VEP screws</td></tr><tr><td>Drill</td><td>Bosch</td><td>Dremel 300 series</td><td>An automatic drill for trepanning</td></tr><tr><td>Electrode lead</td><td>Grass Telefactor&amp;nbsp;</td><td>F-E2-30</td><td>Platinum cables for connecting silver wire electrodes to the amplifier</td></tr><tr><td>Faraday Cage</td><td>custom-made</td><td></td><td>Ensures light proof to maintain dark adaptation. Encloses the Ganzfeld setup to improve signal to noise ratio</td></tr><tr><td>Gauze swabs</td><td>Multigate Medical Products Pty Ltd</td><td>57-100B</td><td>For drying the surgical incision and exposed skull surface during surgery</td></tr><tr><td>Ganzfeld integrating sphere</td><td>Photometric Solutions International</td><td></td><td>Custom designed light stimulator: 36 mm diameter, 13 cm aperture size</td></tr><tr><td>Velcro</td><td>VELCRO Australia Pty Ltd</td><td>VELCRO Brand Reusable Wrap</td><td>Hook-and-loop fastener to secure the electrodes and the animal on the recording platform</td></tr><tr><td>Isoflurane 99.9%</td><td>Abbott Australasia Pty Ltd</td><td>CAS 26675-46-7</td><td>Proprietary Name: Isoflo(TM) Inhalation anaaesthetic. Pharmaceutical-grade inhalation anesthetic mixed with oxygen gas for VEP electrode implant surgery</td></tr><tr><td>Ketamine&amp;nbsp;</td><td>Troy Laboratories</td><td>Ilium Ketamil</td><td>Proprietary name: Ketamil Injection, Brand: Ilium. Pharmaceutical-grade anesthetic for electrophysiological recording</td></tr><tr><td>Luxeon LEDs</td><td>Phillips Lighting Co.</td><td></td><td>For light stimulation twenty 5 W and one 1 W LEDs.</td></tr><tr><td>Micromanipulator</td><td>Harvard Apparatus</td><td>BS4 50-2625</td><td>Holds the ERG active electrode during recordings</td></tr><tr><td>Needle electrode</td><td>Grass Telefactor&amp;nbsp;</td><td>F-E2-30</td><td>Subcutaneously inserted in the tail to serve as the ground electrode for both the ERG and VEP</td></tr><tr><td>Phenylephrine 2.5% minims&amp;nbsp;</td><td>Bausch and Lomb</td><td>CAS 61-76-7</td><td>Instilled with Tropicamide to achieve maximal dilation for ERG recording</td></tr><tr><td>Povidone iodine 10%</td><td>Sanofi-Aventis</td><td>CAS 25655-41-8</td><td>Proprietory name: Betadine, Antiseptic to prepare the shaved skin for surgery 10%, 500 ml</td></tr><tr><td>Powerlab data acquisition system</td><td>ADInstruments</td><td>ML 785</td><td>Controls the LEDs</td></tr><tr><td>Proxymetacaine 0.5%</td><td>Alcon Laboratories&amp;nbsp;</td><td>CAS 5875-06-9</td><td>For corneal anaesthesia during ERG recordings</td></tr><tr><td>Saline solution</td><td>Gelflex</td><td></td><td>Non-injectable, for electroplating silver wire electrodes</td></tr><tr><td>Scope Software</td><td>ADInstruments</td><td>version 3.7.6</td><td>Simultaneously triggers the stimulus via the Powerlab system and collects data</td></tr><tr><td>Silver (fine round wire)</td><td>A&amp;amp;E metal</td><td>0.3 mm</td><td>Used to make active and inactive ERG electrodes, and the inactive VEP electrode</td></tr><tr><td>Stainless streel screws&amp;nbsp;</td><td>MicroFasterners</td><td></td><td>0.7 mm shaft diameter, 3 mm in length to be implanted over the primary visual cortex and serve as the active VEP electrodes</td></tr><tr><td>Stereotaxic frame</td><td>David Kopf</td><td>Model 900</td><td>A small animal stereotaxic instrument for locating the primary visual cortices according to Paxinos &amp;amp; Watson&#039;s 2007 rat brain atlas coordinates</td></tr><tr><td>Surgical blade</td><td>Swann-Morton Ltd.</td><td>0206</td><td>For incising the area of skin overlaying the primary visual cortex to implant the VEP electrodes</td></tr><tr><td>Suture</td><td>Shanghai Pudong Jinhuan Medical Products Co.,Ltd</td><td></td><td>3-0 silk braided suture non-absorbable, for skin retraction during VEP electrode implantation surgery</td></tr><tr><td>Tobramycine eye ointment 0.3%</td><td>Alcon Laboratories&amp;nbsp;</td><td>CAS 32986-56-4</td><td>Proprietary name: Tobrex. Prophylactic antibiotic ointment applied around the skin wound after surgery</td></tr><tr><td>Tropicamide 0.5%</td><td>Alcon Laboratories&amp;nbsp;</td><td>CAS 1508-75-4</td><td>Proprietary name: 0.5% Mydriacyl eye drop, Instilled to achieve mydriasis for ERG recording</td></tr><tr><td>Xylazine</td><td>Troy Laboratories</td><td>Ilium Xylazil-100</td><td>Pharmaceutical-grade anesthetic for electrophysiological recording</td></tr><tr><td>Pipette tip</td><td>Eppendorf Pty Ltd</td><td>0030 073.169</td><td>Eppendorf epTIPS 100 - 5,000 ml, for custom-made electrodes</td></tr><tr><td>Microsoft Office Excel</td><td>Microsoft</td><td>version 2010</td><td>spreadsheet software for data analysis</td></tr><tr><td>Lethabarb Euthanazia Injection</td><td>Virbac (Australia) Pty Ltd</td><td>LETHA450</td><td>325 mg/ml pentobarbital sodium for rapid euthanazia</td></tr></tbody></table>

simultaneous recording of electroretinography and visual evoked potentials in anesthetized rats

The electroretinogram (ERG) and visual evoked potential (VEP) are commonly used to assess the integrity of the visual pathway. The ERG measures the electrical responses of the retina to light stimulation, while the VEP measures the corresponding functional integrity of the visual pathways from the retina to the primary visual cortex following the same light event. The ERG waveform can be broken down into components that reflect responses from different retinal neuronal and glial cell classes. The early components of the VEP waveform represent the integrity of the optic nerve and higher cortical centers. These recordings can be conducted in isolation or together, depending on the application. The methodology described in this paper allows simultaneous assessment of retinal and cortical visual evoked electrophysiology from both eyes and both hemispheres. This is a useful way to more comprehensively assess retinal function and the upstream effects that changes in retinal function can have on visual evoked cortical function.

Die ERG a-Welle (&gt; -1,38 log cd.sm -2), b-Wellen (&gt; - 4,99 log cd.sm -2) STRs (&lt;- 4,99 log cd.sm -2) und die VEP (&gt; - 0.52 log cd.sm -2) wurden gleichzeitig aufgezeichnet (Abbildung 1 und 3). Bei sehr dim Wallungen, eine positive STR ​​(pstr) bei etwa 110 msec nach dem Blitz zu sehen ist , und eine negative STR ​​(nSTR) bei etwa 220 msec (1 und 2). Eine ERG mit einem großen b-Welle, Spitzen zwischen 50 bis 100 ms nach dem Beginn eines moderaten Blitz , die für ihre PII Reaktion analysiert werden können (Abbildungen 1 und 2). An dieser Reizenergie, die negative a-Welle vor dem Gipfel ist vernachlässigbar. Bei helleren Lichtenergien die negative Auslenkung a-Welle wird mehr im Vordergrund , die mit dem PIII - Antwort (2) quantifiziert werden kann. Die scotopic VEP Wellenform zeigt eine negative Antwort (P1N1; 15-70 msec Fenster), gefolgt von<html> eine positive Auslenkung (N1P2; 30-100 msec) (3 und 4). <img alt="Abbildung 1" src="/files/ftp_upload/54158/54158fig1.jpg" /> Abbildung 1. Gruppe Durchschnitt ERG Kurven. Die ERG verändert mit Reizintensität zu erhöhen. Zahlen auf der linken Seite der Wellenform zeigen die Lichtexposition verwendet, um die Wellenform hervorzurufen. Beachten Sie die unterschiedlichen Amplituden und Zeitskalen für jede Platte. Bei Dimmer Leucht Energien die positiven und negativen Komponenten des skotopischen Schwellenreaktion kann ausgelöst werden (pstr, nSTR). Als Stimulus Energien heller zu bekommen, können die a und b-Wellenantwort untersucht werden, und ein gekoppeltes-Flash - Paradigma ermöglicht die Kegel Reaktion gemessen werden. <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54158/54158fig1large.jpg" target="_blank">Bitte hier klicken</a> , um <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54158/54158fig1large.jpg" target="_blank">eine größere Version dieser Figur zu sehen.</a> pload / 54158 / 54158fig2.jpg &quot;/&gt; Abbildung 2. ERG - Analyse. (A) Rod Photorezeptorfunktion kann durch die Verwendung eines PIII getestet werden , um die a-Welle modellieren. A-Wellen bei 1,22 und 1,52 log cd.sm -2 (ungefüllte Kreise, ○) sind fit als Ensemble mit einem PIII (graue Linien, Gleichung 1) auf 90% des Mindestlohns , die Rm PIII (gesättigte Amplitude, &amp; mgr ; V) liefert S (Empfindlichkeit, m 2 .s -1 .cd -3) und td (Zeitverzögerung msec) Parameter. (B) Rod bipolaren Zellfunktion (Mittelwert ± SEM) kann durch Modellieren der Intensität Antwortserie des Stabes PII untersucht werden (leere Kreise ○) mit einer Naka-Rushton-Funktion (graue Linie). Dies gibt V max (gesättigte Amplitude, &amp; mgr ; V), k (1 / Empfindlichkeit, log cd sm -2) und n (Steigung). (C) Retinal Ganglion Zellfunktion bei schwachem Licht Energien untersucht und durch pstr Spitzenamplitude (pstr quantifiziert amp) und Timing (pstr es </sub&gt;). (D) Cone bipolaren Zellfunktion für ein gekoppeltes-Flash - Paradigma quantifiziert durch Kegel PII Spitzenamplitude (Kegel PII amp) und Timing (Kegel PII it) ausgelöst wird. <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54158/54158fig2large.jpg" target="_blank">Bitte klicken Sie hier</a> , um <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54158/54158fig2large.jpg" target="_blank">eine größere Version dieser Figur zu sehen.</a> <img alt="Figur 3" src="/files/ftp_upload/54158/54158fig3.jpg" /> Abbildung 3. Gruppe Durchschnitt VEP Kurven. Die Form der VEP - Wellenform ändert sich mit Reizenergie zu erhöhen. Zahlen auf der linken Seite der Wellenform zeigen die Lichtexposition verwendet , um die Wellenform zu entlocken. <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54158/54158fig3large.jpg" target="_blank">Bitte klicken Sie hier</a> , um <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54158/54158fig3large.jpg" target="_blank">eine größere Version dieser Figur zu sehen.</a> <img alt="Abbildung 4" src="/files/ftp_upload/54158/54158fig4.jpg" /> Figure 4. VEP Analyse und Intensität Antwortfunktion. (A) Amplitudenanalyse des VEP als Spitze genommen Trog (P1N1) und Trog zu Peak (N1P2) Amplituden. Die impliziten Zeiten (es) dieser Antworten wird auch zurückgegeben (P1 es, N1 es, P2 it). (B) Die VEP P1N1 Amplitude (Mittelwert ± SEM) steigt mit Reizenergie zu erhöhen. <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54158/54158fig4large.jpg" target="_blank">Bitte klicken Sie hier</a> , um <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54158/54158fig4large.jpg" target="_blank">eine größere Version zu sehen diese Figur.</a>

Watch this Scientific Journal Video about Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats at JoVE.com

Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats

This protocol describes simultaneous measurement of electroretinogram and visual evoked potentials in anesthetized rats.

Gleichzeitige Aufnahme von Elektroretinographie und visuell evozierten Potentialen in narkotisierten Ratten

The electroretinogram (ERG) and visual evoked potential (VEP) are commonly used to assess the integrity of the visual pathway. The ERG measures the ...

simultaneous-recording-electroretinography-visual-evoked-potentials

Universidad San Sebastian

Research

JoVE Journal

Neuroscience

12.3K Aufrufe.  University of Melbourne. This protocol describes simultaneous measurement of electroretinogram and visual evoked potentials in anesthetized rats. 

Serie: Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats

Implantation and Recording of Wireless Electroretinogram and Visual Evoked Potential in Conscious Rats

The full-field electroretinogram (ERG) and visual evoked potential (VEP) are useful tools to assess retinal and visual pathway integrity in both laboratory and clinical settings. Currently, preclinical ERG and VEP measurements are performed with anesthesia to ensure stable electrode placements. However, the very presence of anesthesia has been shown to contaminate normal physiological responses. To overcome these anesthesia confounds, we develop a novel platform to assay ERG and VEP in conscious rats. Electrodes are surgically implanted sub-conjunctivally on the eye to assay the ERG and epidurally over the visual cortex to measure the VEP. A range of amplitude and sensitivity/timing parameters are assayed for both the ERG and VEP at increasing luminous energies. The ERG and VEP signals are shown to be stable and repeatable for at least 4 weeks post surgical implantation. This ability to record ERG and VEP signals without anesthesia confounds in the preclinical setting should provide superior translation to clinical data.

We show surgical implantation and Recording procedures to measure visual electrophysiological signals from the eye (electroretinogram) and brain (visual evoked potential) in conscious rats, which is more analogous to the human condition where recordings are conducted without anesthesia confounds.

Implantations- und Aufzeichnung von Wireless-Elektroretinogramm und visuell evozierten Potenziale in wachen Ratten

The full-field electroretinogram (ERG) and visual evoked potential (VEP) are useful tools to assess retinal and visual pathway integrity in both ...

Forschung

Verhalten

Using the Electroretinogram to Assess Function in the Rodent Retina and the Protective Effects of Remote Limb Ischemic Preconditioning

The ERG is the sum of all retinal activity. The ERG is usually recorded from the cornea, which acts as an antenna that collects and sums signals from the retina. The ERG is a sensitive measure of changes in retinal function that are pan-retinal, but is less effective for detecting damage confined to a small area of retina. In the present work we describe how to record the &#8216;flash&#8217; ERG, which is the potential generated when the retina is exposed to a brief light flash. We describe methods of anaesthesia, mydriasis and corneal management during recording; how to keep the retina dark adapted; electrode materials and placement; the range and calibration of stimulus energy; recording parameters and the extraction of data. We also describe a method of inducing ischemia in one limb, and how to use the ERG to assess the effects of this remote-from-the-retina ischemia on retinal function after light damage. A two-flash protocol is described which allows isolation of the cone-driven component of the dark-adapted ERG, and thereby the separation of the rod and cone components. Because it can be recorded with techniques that are minimally invasive, the ERG has been widely used in studies of the physiology, pharmacology and toxicology of the retina. We describe one example of this usefulness, in which the ERG is used to assess the function of the light-damaged retina, with and without a neuroprotective intervention; preconditioning by remote ischemia.

The electroretinogram (ERG) is an electrical potential generated by the retina in response to light. This paper describes how to use the ERG to assess retinal function, in dark-adapted rats, and how it can be can be used to assess a neuroprotective intervention, in the present case remote ischemic preconditioning.

Verwenden der Funktion zur Elektroretinogramm im Nager Retina und die schützende Wirkung des Fern Limb ischämischen Präkonditionierung bewerten

The ERG is the sum of all retinal activity. The ERG is usually recorded from the cornea, which acts as an antenna that collects and sums signals from the ...

Neurowissenschaften

Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to monitor the disease course in the follow-up period. Changes in the VEP parameters closely correlate with pathological damage in the optic nerve. This protocol provides a detailed description about the rodent model of optic nerve microinjection, in which a partial demyelination lesion is produced in the optic nerve. VEP recording techniques are also discussed. Using skull implanted electrodes, we are able to acquire reproducible intra-session and between-session VEP traces. VEPs can be recorded on individual animals over a period of time to assess the functional changes in the optic nerve longitudinally. The optic nerve demyelination model, in conjunction with the VEP recording protocol, provides a tool to investigate the disease processes associated with demyelination and remyelination, and can potentially be employed to evaluate the effects of new remyelinating drugs or neuroprotective therapies.

Focal demyelination is induced in the optic nerve using lysolecithin microinjection. Visual evoked potentials are recorded via skull electrodes implanted over the visual cortex to examine the signal conduction along the visual pathway in vivo. This protocol details the surgical procedures underlying electrode implantation and optic nerve microinjection.

Visuell evozierten Potential-Aufnahme in einem Rattenmodell für Experimentelle Optic Nerve Demyelinisierung

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to ...

Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent

Although electroencephalography (EEG) is widely used as a non-invasive technique for recording neural activities of the brain, our understanding of the neurogenesis of EEG is still very limited. Local field potentials (LFPs) recorded via a multi-laminar microelectrode can provide a more detailed account of simultaneous neural activity across different cortical layers in the neocortex, but the technique is invasive. Combining EEG and LFP measurements in a pre-clinical model can greatly enhance understanding of the neural mechanisms involved in the generation of EEG signals, and facilitate the derivation of a more realistic and biologically accurate mathematical model of EEG. A simple procedure for acquiring concurrent and co-localized EEG and multi-laminar LFP signals in the anesthetized rodent is presented here. We also investigated whether EEG signals were significantly affected by a burr hole drilled in the skull for the insertion of a microelectrode. Our results suggest that the burr hole has a negligible impact on EEG recordings.

This protocol describes a simple method for concurrent recording of co-localized electroencephalography (EEG) and multi-laminar local field potential in an anesthetized rat. A burr hole drilled in the skull for the insertion of a microelectrode is shown to produce negligible distortion of the EEG signal.

Gleichzeitige Aufnahme von Co lokalisierte Elektroenzephalographie und lokales Feld Potential in Nagetier

Although electroencephalography (EEG) is widely used as a non-invasive technique for recording neural activities of the brain, our understanding of the ...

Electroretinogram Recording for Infants and Children under Anesthesia to Achieve Optimal Dark Adaptation and International Standards

Electroretinogram (ERG) is the only clinical objective test available to assess retinal function. Full-field ERG (ffERG) measures the panretinal rod and cone photoreceptor function as well as inner retinal function and is an important measure in the diagnosis and management of inherited retinal diseases as well as inflammatory, toxic, and nutritional retinopathies. Adhering to international standards and maintaining retinal dark adaptation are critical to acquire valid and reliable dark-adapted (scotopic) and light-adapted (photopic) ffERG responses. Performing ffERG in infants and children is challenging and often requires general anesthesia in the operating room. However, maintaining retinal dark adaptation in the operating room is becoming increasingly difficult given the numerous light sources from anesthesiology monitoring systems and other equipment. A practical and widely applicable method for ffERG testing is described in the operating room that optimizes retinal dark adaptation. The method reduces operating room time by dark-adapting the patient before general anesthesiology is instituted. The operating room is modified for dark adaptation and any remaining light source in the darkened operating room is minimized with the use of a modified portable foldable darkroom that encloses the patient&#8217;s head and the ERG examiner during ffERG scotopic recordings. The simple method adheres to ffERG international standards and provides valid reliable scotopic and photopic ffERG recordings that are critical to assess objective retinal function in this young age group where subjective assessment of visual function such as visual acuity and visual fields are not possible. Furthermore, the ffERG is the gold standard clinical test in detecting early onset inherited retinal diseases including Leber congenital amaurosis where approved gene therapy has become available. In sedated conditions, very low amplitude ffERG signals can be detected due to minimal orbicularis muscle activity interference, which is particularly relevant in patients after gene therapy to detect improved amplitude responses.

Adhering to international standards and maintaining retinal dark adaptation are critical to acquire valid full-field electroretinogram responses in the diagnosis and management of inherited retinal diseases. A practical protocol using a portable darkroom is provided to obtain full-field&#160;electroretinogram for infants and children under sedation or general anesthesia in the operating room setting.

Electroretinogram Recording für Säuglinge und Kinder unter Anästhesie, um optimale dunkle Anpassung und internationale Standards zu erreichen

Electroretinogram (ERG) is the only clinical objective test available to assess retinal function. Full-field ERG (ffERG) measures the panretinal rod and ...

Medizin

Full-Circle Cauterization of Limbal Vascular Plexus for Surgically Induced Glaucoma in Rodents

Glaucoma, the second leading cause of blindness worldwide, is a heterogeneous group of ocular disorders characterized by structural damage to the optic nerve and retinal ganglion cell (RGC) degeneration, resulting in visual dysfunction by interrupting the transmission of visual information from the eye to the brain. Elevated intraocular pressure is the most important risk factor; thus, several models of ocular hypertension have been developed in rodents by either genetic or experimental approaches to investigate the causes and effects of the disease. Among those, some limitations have been reported such as surgical invasiveness, inadequate functional assessment, requirement of extensive training, and highly variable extension of retinal damage. The present work characterizes a simple, low-cost, and efficient method to induce ocular hypertension in rodents, based on low-temperature, full-circle cauterization of the limbal vascular plexus, a major component of aqueous humor drainage. The new model provides a technically easy, noninvasive, and reproducible subacute ocular hypertension, associated with progressive RGC and optic nerve degeneration, and a unique post-operative clinical recovery rate that allows in vivo functional studies by both electrophysiological and behavioral methods.

The goal of this protocol is to characterize a novel model of glaucomatous neurodegeneration based on 360&#176; thermic cauterization of limbal vascular plexus, inducing subacute ocular hypertension.

Vollkreiskauterisierung des limbalen Gefäßplexus bei chirurgisch induziertem Glaukom bei Nagetieren

Glaucoma, the second leading cause of blindness worldwide, is a heterogeneous group of ocular disorders characterized by structural damage to the optic ...

Preparing a Rat Model to Assess Visual Pathway Integrity Using an Electrophysiology Setup

Source: Nguyen, C. T. et al., <a href="https://app-jove-com.bdigitaluss.remotexs.co/v/54158">Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats.</a>&#160;J. Vis. Exp. (2016)
This video demonstrates the preparation of a rat model to study retinal health and visual pathway integrity. Visual evoked potential and electroretinography reference and active electrodes are connected to the rat, followed by aligning the platform with the rat in front of the Ganzfeld bowl for even retinal illumination and closing the Faraday cage to avoid external interference.

Vorbereitung eines Rattenmodells zur Beurteilung der Integrität der Sehbahn unter Verwendung eines elektrophysiologischen Aufbaus

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

JoVE Encyclopedia of Experiments

Neurophysiology

Electrophysiology Techniques

Direct-Coupled Electroretinogram Recording of the Retinal Pigment Epithelium in a Mouse

Source: Miyagishima, K. J., et al. <a href="https://app-jove-com.bdigitaluss.remotexs.co/v/61491">Direct-Coupled Electroretinogram (DC-ERG) for Recording the Light-Evoked Electrical Responses of the Mouse Retinal Pigment Epithelium.</a> J. Vis. Exp. (2020).
This video demonstrates the recording of a direct-coupled electroretinogram (DC-ERG) on a dark-adapted, anesthetized mouse retinal pigment epithelium (RPE). After applying dilating drops, the animal is placed in a recording setup, and the capillary electrode filled with a salt buffer is placed on the cornea. After establishing the baseline, DC-ERG is recorded.

Direkt gekoppeltes Elektroretinogramm Aufzeichnung des retinalen Pigmentepithels bei einer Maus

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Mouse ...

Imaging and Optical Techniques

Recording of Visual Evoked Potentials via the Skull Electrodes in a Rat Model

Source: You, Y., et al. <a href="https://app-jove-com.bdigitaluss.remotexs.co/v/52934">Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination.</a> J. Vis. Exp. (2015).
This video demonstrates the process of measuring visual evoked potentials or VEPs in a dark-adapted, anesthetized rat with pre-implanted skull electrodes, which record the electrical signals from the brain&#39;s visual cortex in response to light stimuli.

Aufzeichnung visuell evozierter Potentiale über die Schädelelektroden in einem Rattenmodell

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. VEP ...

Concurrent Electroencephalographic and Local Field Potential Recordings in an Anesthetized Rat

Source: Kang, S., et al. <a href="https://app-jove-com.bdigitaluss.remotexs.co/v/56447">Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent.</a> J. Vis. Exp. (2017).
This video demonstrates the procedure for concurrent recordings of co-localized electroencephalography (EEG) and local field potentials (LFPs) in an anesthetized rat. The process involves preparing the rat and attaching stimulating electrodes to its whisker pads. An EEG spider electrode is placed on the rat&#39;s skull for surface EEG measurements. At the same time, a multi-channel microelectrode is inserted into a specific brain region through a burr hole in the skull to record neural activity from a localized brain area.