Name: implantation and recording of wireless electroretinogram and visual evoked potential in conscious rats
Uploaded: 2016-06-29T15:00:00
Description: Watch this Scientific Journal Video about Implantation and Recording of Wireless Electroretinogram and Visual Evoked Potential in Conscious Rats at JoVE.com

JC would like to acknowledge the David Hay Memorial Fund, The University of Melbourne for financial support in writing this manuscript. Funding for this project was provided by an ARC Linkage grant 100200129 (BVB, AJV, CTON).

Etik uttalande: Djurförsök genomfördes i enlighet med den australiensiska koden för skötsel och användning av djur för vetenskapliga ändamål (2013). Djuretik godkännande erhölls från Animal etikkommitté, University of Melbourne. Materialen häri är för laboratorieförsök, och inte är avsedda för medicinskt eller veterinärt bruk. 1. Förbereda Elektroder Obs: Tre kanalsändare används för kirurgisk implantation som möjliggör två ERG och en VEP inspelning ska genomfö...

<ol>
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På grund av den minimalinvasiv karaktär visuell elektro är ERG och VEP inspelningar i humanpatienter genomförs under medvetna förhållanden och endast kräver användning av lokalbedövningsmedel för elektrodplacering. Däremot är visuell elektro i djurmodeller konventionellt utförs under narkos för att möjliggöra en stabil elektrodplacering genom att eliminera frivilliga ögon och kroppsrörelser. Men vanligen använda narkos ändra ERG och VEP svar som framgår av vår tidigare publikation 24 och...

ERG och VEP är minimalt invasiva in vivo verktyg för att bedöma integriteten av retinala och visuella vägar respektive i både laboratoriet och kliniken. Den fullständiga fält ERG ger en karakteristisk vågform som kan brytas ner i olika komponenter, med varje element representerar olika cellklasser näthinnans vägen 1,2. Den klassiska fullständiga fältet ERG vågform består av en initial negativ lutning (a-vågen), vilket har visat sig representera fotoreceptoraktivitet inlägget ljusexponering 2-4. A-vågen följs av en betydande positiv vågform (b-vågen) som avspeglar elektrisk aktivitet i medel näthinnan, främst de ON-bipolära celler 5-7. Dessutom kan man variera ljusenergi och inter stimulus-intervall för att isolera kon från stav svar 8. Blixten VEP representerar elektriska potentialerna syncentrum och hjärnstammen som svar på näthinnan ljus stimulering9,10. Denna vågform kan delas upp i tidiga och sena komponenter, med den tidiga komponenten reflekterande aktivitet av nervceller i Retino-geniculo-striate väg 11-13 och den sena komponenten representerar kortikala behandlingen som utförs i olika V1 lamellerna hos råttor 11,13. Därför samtidig mätning av ERG och VEP returnerar omfattande bedömning av de strukturer som deltar i den visuella vägen. För närvarande, för att spela in elektrofysiologi i djur, är anestesi används för att möjliggöra en stabil placering av elektroderna. Det har gjorts försök att mäta ERG och VEP i medvetna råttor 14-16 men dessa studier användes en fast installation, som kan vara besvärligt och kan leda till djur stress genom att begränsa djurförflyttningar och naturliga beteende 17. Med de senaste framstegen inom trådlös teknik, inklusive förbättrad miniatyrisering och batteritid, är det nu möjligt att genomföra en telemetri strategi för ERG end VEP inspelning, minskar stress i samband med trådbundna inspelningar och förbättra lång sikt. Fullt interna stabila implantationer av telemetri sonder har visat sig vara framgångsrik för kronisk övervakning av temperatur, blodtryck 18, aktivitet 19 liksom elektroencefalografi 20. Sådana tekniska framsteg kommer också att hjälpa med repeterbarhet och stabilitet vid medvetande inspelningar, öka plattformens verktyg för kroniska studier.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Bioamplifier</td>
			<td>ADInstruments</td>
			<td>ML 135</td>
			<td>Amplifies ERG and VEP signals</td>
		</tr>
		<tr>
			<td>Carboxymethylcellulose sodium 1.0%</td>
			<td>Allergan</td>
			<td>CAS 0009000-11-7</td>
			<td>Maintain corneal hydration during surgery</td>
		</tr>
		<tr>
			<td>Carprofen 0.5%</td>
			<td>Pfizer Animal Health Group</td>
			<td>CAS 53716-49-7</td>
			<td>Post-surgery analgesia, given with injectable saline for fluid replenishment</td>
		</tr>
		<tr>
			<td>Chlorhexadine 0.5%</td>
			<td>Orion Laboratories</td>
			<td>27411, 80085</td>
			<td>Disinfection of surgical instrument</td>
		</tr>
		<tr>
			<td>Cyanoacrylate gel activator</td>
			<td>RS components</td>
			<td>473-439</td>
			<td>Quickly dries cyanoacrylate gel</td>
		</tr>
		<tr>
			<td>Cyanocrylate gel&#160;</td>
			<td>RS components</td>
			<td>473-423</td>
			<td>Fix stainless screws to skull</td>
		</tr>
		<tr>
			<td>Dental burr</td>
			<td>Storz Instruments, Bausch and Lomb</td>
			<td>E0824A</td>
			<td>Miniature drill head of ~0.7mm 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>Automatic drill for trepanning</td>
		</tr>
		<tr>
			<td>Enrofloxin</td>
			<td>Troy Laboratories</td>
			<td></td>
			<td>Prophylactic antibiotic post surgey</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>Gauze swabs</td>
			<td>Multigate Medical Products Pty Ltd</td>
			<td>57-100B</td>
			<td>Dries surgical incision and exposed skull surface during surgery</td>
		</tr>
		<tr>
			<td>Isoflurane 99.9%</td>
			<td>Abbott Australasia Pty Ltd</td>
			<td>CAS 26675-46-7</td>
			<td>Proprietory Name: Isoflo(TM) Inhalation anaaesthetic. Pharmaceutical-grade inhalation anesthetic mixed with oxygen gas for VEP electrode implant surgery</td>
		</tr>
		<tr>
			<td>Kenacomb ointment</td>
			<td>Aspen Pharma Pty Ltd</td>
			<td></td>
			<td>To reduce skin irritation and itching after surgery</td>
		</tr>
		<tr>
			<td>Luxeon LEDs</td>
			<td>Phillips Lighting Co.</td>
			<td></td>
			<td>For light stimulation, twenty 5 watt and one 1 watt LEDs, controlled by Scope software</td>
		</tr>
		<tr>
			<td>Needle (macrosurgery)</td>
			<td>World Precision Instruments</td>
			<td>501959</td>
			<td>for suturing abdominal and head surgery, used with 3-0 suture, eye needle, cutting edge 5/16 circle Size 1, 15mm</td>
		</tr>
		<tr>
			<td>Needle holder (macrosurgery)</td>
			<td>World Precision Instruments</td>
			<td>500224</td>
			<td>To hold needle during abdominal and head surgery</td>
		</tr>
		<tr>
			<td>Needle holder (microsurgery)</td>
			<td>World Precision Instruments</td>
			<td>555419NT</td>
			<td>To hold needle during ocular surgery</td>
		</tr>
		<tr>
			<td>Optiva catheter</td>
			<td>Smiths Medical International LTD</td>
			<td>16 or 21 G</td>
			<td>Guide corneal active electrodes from skull to conjunctiva</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>Acquire signal from telemetry transmitter, paired to telemetry data converter</td>
		</tr>
		<tr>
			<td>Proxymetacaine 0.5%</td>
			<td>Alcon Laboratories&#160;</td>
			<td>CAS 5875-06-9</td>
			<td>Topical ocular analgesia</td>
		</tr>
		<tr>
			<td>Restrainer</td>
			<td>cutom made</td>
			<td></td>
			<td>Front of the restrainer is tapered to minimize head movement, length can be adjusted to accommodate different rat length, overall diameter is 60 mm.&#160;</td>
		</tr>
		<tr>
			<td>Scapel blade</td>
			<td>R.G. Medical Supplies</td>
			<td>SNSM0206</td>
			<td>For surgical incision</td>
		</tr>
		<tr>
			<td>Scissors (macrosurgery)</td>
			<td>World Precision Instruments</td>
			<td>501225</td>
			<td>for cutting tissue on the abodmen and forhead</td>
		</tr>
		<tr>
			<td>Scissors (microsurgery)</td>
			<td>World Precision Instruments</td>
			<td>501232</td>
			<td>To dissect the conjunctiva for electrode attachment</td>
		</tr>
		<tr>
			<td>Scope Software</td>
			<td>ADInstruments</td>
			<td>version 3.7.6</td>
			<td>Simultaneously triggers the stimulus via the ADI Powerlab system and collects data</td>
		</tr>
		<tr>
			<td>Shaver</td>
			<td>Oster</td>
			<td>Golden A5</td>
			<td>Shave fur from surgical areas</td>
		</tr>
		<tr>
			<td>Stainless streel screws&#160;</td>
			<td>MicroFasteners</td>
			<td>L001.003CS304</td>
			<td>0.7 mm shaft diameter, 3 mm in length&#160;</td>
		</tr>
		<tr>
			<td>Stereotaxic frame</td>
			<td>David Kopf</td>
			<td>Model 900</td>
			<td>A small animal stereotaxic instrument for locating the implantation landmarks on the skull</td>
		</tr>
		<tr>
			<td>Surgical drape</td>
			<td>Vital Medical Supplies</td>
			<td>GM29-612EE</td>
			<td>Ensure sterile enviornment during surgery</td>
		</tr>
		<tr>
			<td>Suture (macrosurgery)</td>
			<td>Ninbo medical needles</td>
			<td>3-0</td>
			<td>for suturing abdominal and head surgery, sterile silk braided, 60cm</td>
		</tr>
		<tr>
			<td>Suture needle (microsurgery)</td>
			<td>Ninbo medical needles</td>
			<td>8-0 or 9-0</td>
			<td>for ocular surgery including, suturing electrode to sclera and closing conjunctival wound, nylon suture, 3/8 circle 1&#215;5, 30cm</td>
		</tr>
		<tr>
			<td>Telemetry data converter&#160;</td>
			<td>DataSciences International</td>
			<td>R08</td>
			<td>allows telemetry signal to interface with data collection software</td>
		</tr>
		<tr>
			<td>Telemetry Data Exchange Matrix</td>
			<td>DataSciences International</td>
			<td></td>
			<td>Gathers data from transmitters, pair with receiver</td>
		</tr>
		<tr>
			<td>Telemetry data receiver</td>
			<td>DataSciences International</td>
			<td>RPC-1</td>
			<td>Receives telemetry data from transmitter</td>
		</tr>
		<tr>
			<td>Telemetry transmitter</td>
			<td>DataSciences International</td>
			<td>F50-EEE</td>
			<td>3 channel telemetry transmitter</td>
		</tr>
		<tr>
			<td>Tropicamide 0.5%</td>
			<td>Alcon Laboratories&#160;</td>
			<td></td>
			<td>Iris dilation</td>
		</tr>
		<tr>
			<td>Tweezers (macrosurgery)</td>
			<td>World Precision Instruments</td>
			<td>500092</td>
			<td>Manipulate tissues during abdominal and head surgery</td>
		</tr>
		<tr>
			<td>Tweezers (microsurgery)</td>
			<td>World Precision Instruments</td>
			<td>500342</td>
			<td>Manipulate tissues during ocular surgery</td>
		</tr>
	</tbody>
</table>

implantation and recording of wireless electroretinogram and visual evoked potential in conscious rats

Den fullständiga fält elektroretinogram (ERG) och visuellt framkallat potential (VEP) är användbara verktyg för att bedöma retinal och visuella vägen integritet i både laboratorie- och kliniska miljöer. För närvarande är prekliniska ERG och VEP mätningar utfördes med anestesi att säkerställa stabila elektrodplaceringar. Emellertid har mycket närvaro av anestesi visats förorena normala fysiologiska responser. För att övervinna dessa anestesi blandar ihop, utvecklar vi en ny plattform för att analysera ERG och VEP i råttor vid medvetande. Elektroder inopererade under conjunctivally på ögat för att analysera ERG och epiduralt över syncentrum för att mäta VEP. En rad amplitud och känslighet / tidsparametrar analyseras för både ERG och VEP öka självlysande energier. ERG och VEP signaler visas sig vara stabil och repeterbar under åtminstone 4 veckor efter kirurgisk implantation. Denna förmåga att spela ERG och VEP signaler utan bedövning confounds i prekliniska sKomma bör ge överlägsen översättning till kliniska data.

Fotoreceptorsvar analyseras genom att montera en fördröjd Gauss till framkanten av den första fallande delen av ERG svar på de översta 2 lysande energier (1,20, 1,52 log csm -2) för varje djur, baserat på modellen av lamm och Pugh 22, utarbetat av Hood och Birch 23. Denna formel returnerar en amplitud och en känslighetsparameter (Figur 1C och 1D, respektive). En hyperbolisk funktion anpassades till ljusenergin sva...

Watch this Scientific Journal Video about Implantation and Recording of Wireless Electroretinogram and Visual Evoked Potential in Conscious Rats at JoVE.com

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

Vi visar kirurgiska implantation och inspelning förfaranden för att mäta visuella elektrofysiologiska signaler från ögat (elektroretinogram) och hjärna (visuell evoked potential) hos råttor vid medvetande, vilket är mer analogt med det mänskliga tillståndet där inspelningar görs utan bedövning blandar ihop.

Implantation och inspelning av Wireless elektroretinogram och Visual Evoked Potential hos råttor vid medvetande

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

The overall goal of this procedure is to measure conscious ERG and VEP signals following surgical implantation of wireless telemetry probes. This technique can help answer key questions in the region of neuroscience field, such as how neurons behave without the limitation of general anesthesia under both healthy conditions and diseased state. The main advantage of this technique is that it allows simultaneous recording of the electro physiology for the retina and for the visual cortex in conscious animal.To begin this procedure prepare the electrodes under a microscope. To do so untwist a double stranded, stainless steel electrode with two fine tipped pliers. Trim one of the stainless steel strands at approximately one centimeter from the tip, leaving a single longer, straight strand to shape the ring electrode.Then, fold the single stainless steel strand back into it self and twist to form a smooth ring at the tip of the electrode. Next shape an ERG active electrode by twisting the base of the loop to form a circle with a diameter of point two to point five millimeters. For the two ERG inactive and one VEP inactive electrodes make the loop diameter to point eight millimeters.In this example, prepare one active ERG and three inactive electrodes. After that, hook the circular VEP active electrode around a stainless steel screw so the electrode rests against the screw head. Then, hook the three inactive electrodes around a second stainless steel screw.The ground electrode will go directly into the peritoneal cavity and there is no need to make a loop at the tip. After shaving and disinfecting the surgical site of an anesthetized animal, make a 10 millimeter incision on the head along he mid line between the ears with a surgical scalpel. Then, make a five millimeter incision on the abdomen through the skin layer, along the mid line below the sternum.After that, tunnel a cannula of five millimeter diameter subcutaneously, from the abdomen incision to the head incision. Feed the electrode wires of the tansmitter through the cannula from the abdomen to the head. Leave the reference electrode with the transmitter base.Then, cover all the electrode tips with aseptic gauze. Next, secure the rat's head to a stereotaxic platform. Extend the forehead incision to 30 millimeters with surgical scissors.Then, expose a surgical area by retracting the skin with two sutures. Scrape off the periosteum overlying the skull to expose the bregma, lambda and mid line sutures. Subsequently, drill two holes through the skull at the VEP active and ERG, VEP inactive stereotaxic coordinates.Attach the active and inactive electrodes with stainless steel screws to the skull to about one millimeter in depth. To implant the ERG active electrodes, use a suture to temporarily retract the upper eye lid. Then, insert a 16 to 21 gauge cannula subcutaneously from behind the eye through the superior conjunctival fornix to the forehead.Subsequently, remove the guiding needle. Next, feed the active electrode through plastic catheter from the forehead towards the eye. Then, remove the plastic catheter.Use a temporary suture, which is threaded through the electrode loop to prevent the electrode from retracting back into the tunnel. After that, make a point five millimeter incision on the superior conjunctiva one millimeter behind the limbus. Use blunt dissection to expose the underlying sclera.Then, implant a suture immediately behind the limbus at half scleral thickness and be careful not to puncture the eyeball. Subsequently, remove the temporary suture from the ERG active electrode. Anchor the ERG active electrode to the half scleral thickness suture by tying three consecutive knots and ensure the electrode tip is situated close to the limbus for good signal to noise ratio.If necessary pull back the excess electrode lead so the active electrode doesn't impinge on the cornea. Then, close the conjuntival flap using one to two interrupted sutures to completely cover the ERG electrode. Afterward, remove the eyelid retracting suture.Repeat the procedure for the contralateral eye. Now, apply cyanoacrylate gel over the skull to secure all the stainless steel screws and electrode wires. Ensure the ERG active electrodes are not tightly pulled before being secured to enable eye movements.Next, close the incision using interrupted sutures. Then, rotate the rodent to expose the abdominal area. Extend the abdominal dermal incision to 40 millimeters along the linea alba with surgical scissors.After that, make a 35 millimeter incision through the inner muscle wall, to expose the inner abdominal cavity. Using two sutures, attach the transmitter body to the animal's inner abdominal wall at the right and avoid contacting the liver. Fold the ground electrode by securing it in a loop with two sutures and place it, free floating, in the abdominal cavity along with the other electrode leads.Subsequently, close the peritoneum using a continuous suture. Then close the skin incision using interrupted sutures. For recording dark adapted signals animals are dark adapted for 12 hours and all procedures are preformed in a dark room with the aide of dim red light.For illustration purposes recordings will be conducted under normal room lighting conditions here. Before recording, apply topical anesthesia and dilating drops to the cornea of the rat. Next, guide the conscious animal into a custom made, clear restrainer.Place the rat in front of the Ganzfeld bowl with it's eyes aligned with the opening of the bowl. Turn on the indwelling transmitter by passing a magnet within five centimeters of it. Verify that the transmitter is on by checking the LED status light on the receiver base.Close the Faraday cage which house's the receiver pad to improve the signal to noise ratio. Then, collect signals over a range of luminous energies. This figure shows the ERG wave forms in conscious rats on day seven post surgery.ERG responses were recorded to range of stimulus energy with the response to the dim flash shown at the bottom and the brightest flash at the top. On day 28 post surgery, these wave forms remained similar to those on day seven. Over four weeks there was no significant time effect for photo receptor amplitude and sensitivity.Rod bi polar cell amplitude and sensitivity. Cone bi polar cell amplitude. And implicit time.Similarly, VEP wave forms appear to be comparable seven and 28 days post surgery. With amplitude and timing parameters showing no significant time effect. These results indicated robust ERG and VEP signal stability.Once mastered, the surgical component of the technique will be completed in one and half to two hours. The conscious recording component, if preformed properly can be completed in 45 minutes. Following this procedure, other methods like pharmacological or disease manipulation can also be preformed.This will answer additional questions in drug discovery as well as chronicle longitudinal experiments. Additionally this procedure can also be preformed in other animals if required. After watching this video, you should have a good understanding of how to surgically implant and subsequently measure conscious ERG and VEP telemetry signals.

Research

JoVE Journal

Behavior

The Successive Alleys Test of Anxiety in Mice and Rats

De successiva Gränderna Test av ångest hos möss och råttor

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones.  ...

Automated Visual Cognitive Tasks for Recording Neural Activity Using a Floor Projection Maze

Automatiserade Visual kognitiva uppgifter för inspelning Neural aktivitet Använda en Floor Projection Maze

Neuropsychological tasks used in primates to investigate mechanisms of learning and memory are typically visually guided cognitive tasks. We have ...

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

Den modifierade Hole Board - Mätning Beteende, Kognition och social interaktion i möss och råttor

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

Conscious and Non-conscious Representations of Emotional Faces in Asperger's Syndrome

Medvetna och icke-medvetna representationer av emotionella ansikten i Aspergers syndrom

Several neuroimaging studies have suggested that the low spatial frequency content in an emotional face mainly activates the amygdala, pulvinar, and ...

Manipulation of Epileptiform Electrocorticograms (ECoGs) and Sleep in Rats and Mice by Acupuncture

Manipulation of Epileptiform Electrocorticograms ECoGs and Sleep in Rats and Mice by Acupuncture

Manipulering av Epileptiforma Electrocorticograms (ECoGs) och sömn hos råttor och möss genom akupunktur

Ancient Chinese literature has documented that acupuncture possesses efficient therapeutic effects on epilepsy and insomnia. There is, however, little ...

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

En trådlös, dubbelriktad Interface för In Vivo inspelning och stimulering av Neural aktivitet i fritt bete råtta

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

Using Electroencephalography Measurements and High-quality Video Recording for Analyzing Visual Perception of Media Content

Med hjälp av elektroencefalografi mätningar och hög kvalitet videoinspelning för att analysera visuell Perception av medieinnehåll

This article explores a method to detect differences in visual perception in humans. The method used is based on the psychological (or "cognitive") ...

Eye Tracking During Visually Situated Language Comprehension: Flexibility and Limitations in Uncovering Visual Context Effects

Eye Tracking under visuellt ligger språkförståelse: Flexibilitet och begränsningar i avslöjande visuella sammanhang effekter

The present work is a description and an assessment of a methodology designed to quantify different aspects of the interaction between language processing ...

Visual Classical Conditioning in Wood Ants

Visuella klassisk betingning i trä myror

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, ...

Investigating the Deployment of Visual Attention Before Accurate and Averaging Saccades via Eye Tracking and Assessment of Visual Sensitivity

Utreda utplaceringen av visuell uppmärksamhet innan korrekt och genomsnitt Saccades via Eye Tracking och bedömning av visuella känslighet

This experimental protocol was designed to investigate whether visual attention is obligatorily deployed at the endpoint of saccades. To this end, we ...