Name: simultaneous recording of electroretinography and visual evoked potentials in anesthetized rats
Uploaded: 2016-07-01T14:00:00
Description: Watch this Scientific Journal Video about Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats at JoVE.com

Funding for this project was provided by the National Health and Medical Research Council (NHMRC) 1046203 (BVB, AJV) and Melbourne Neuroscience Institute Fellowship (CTN).

모든 실험 절차는 호주 국립 보건 의료 연구위원회에 의해 규정 된 관리 및 과학적인 목적을위한 동물의 사용을위한 연습 호주 코드에 따라 실시 하였다. 윤리 통관은 멜버른 대학 과학 학부, 동물 윤리위원회 (승인 번호 0911322.1)로부터 얻은 것입니다. 만성 VEP 전극 1. 사전 주입 주 : 동시 ERG와 VEP 신호가 수술 VEP 이식해야한다 동물은 적어도 일주 컬렉션 신호 전에 전극 수집 할 경우. </p...

<ol>
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르와 VEP는 각각 망막 및 피질에서 시각 기능의 객관적 측정합니다. 동시 기록의 장점은 전체 시각적 경로의보다 포괄적 인보기가 수득된다는 점이다. 특히, 자신의 동시 평가에서 보완적인 정보는 예를 들어, 시신경 병증 차 뇌 위축 (19)와 공존 할 수 ERG 아직 뚜렷한 VEP 18 발현 중복과 장애에 대한 (시각 경로에 부상의 사이트의 명확한 묘사를 제공 할 수있다, 20, 또?...

전위도 (ERG) 및 시각 유발 전위 (VEP)의 측정은 시각 경로의 무결성의 유용한 정량적 평가를 제공합니다. VEP이 같은 빛 이벤트 다음 차 시각 피질에 망막에서 시각 경로의 해당 기능 무결성을 측정하면서 ERG는 빛 자극에 망막의 전기 반응을 측정한다. 이 원고는 일반적으로 사용되는 실험실 모델 쥐의 기록 및 ERG와 VEP 응답의 분석을위한 프로토콜을 설명합니다. 르 빛의 플래시에 망막의 총 전기 반응을 정량화하여 키 망막 세포 클래스의 숫자의 기능 무결성의 인덱스를 제공합니다. 이온 플럭스의 조율 된 일련의 광 개시에 의해 시작 오프셋, 눈의 외부 표면에 배치 된 전극을 사용하여 측정 할 수있는 전압 검출 가능한 변화를 생성한다. 얻어진 파형은 자체의 조합을 나타내는진폭, 타이밍 및 주파수의 다른 잘 정의 된 컴포넌트 RIES. 연구의 실질적인 몸체는 이러한 성분이 비교적 많은 척추 동물 망막 통해 상기 구성 요소가 서로로부터 분리 될 수 있다는 것을 잘 보존되어 있는지 보여 주었다. 적절하게 자극 (플래시 자극, 배경, interstimulus 간격) 조건을 선택하고 분석하는 복합 파형의 특정 기능을 선택하여 하나의 망막 세포 1,2의 특정 그룹의 측정 값을 반환 확신 할 수 있습니다. 이러한 특성은 유틸리티 따라서 망막 기능의 비 침습적 척도로서 ERG의 광범위한 애플리케이션을 기초. 이 원고는 (긍정적를 ERG을 측정하고 망막의 주요 세포 종류, 즉 광 수용체합니다 (PIII 구성 요소), 양극성 세포 (개인 식별 구성 요소) 및 망막 신경절 세포의 일부에 대한 정보를 반환하는 데 그 특징을 분석하는 방법론에 초점을 맞추고 암순응 임계 응답 또는 pSTR). 3의 영역 V1에 시신경, 시신경 기관, 시상 (측면 geniculate 핵, LGN) 및 광섬유 방사선을 통해 직렬 통신. 설치류에서, 다수 - 각 눈 decussate 4에서 시신경 섬유 (90-95 %)과는 반대측의 중간 뇌에 분포. 르 달리 5 따라서 시각적 통로가 VEP 파형에 영향을 미칠 수있는 어느 곳 따라 변경 특정 세포 종류에 VEP의 다른 구성 요소를 돌리는 아직 가능하지 않다. 그럼에도 불구하고, VEP 시각적 성능 및 시각적 경로 무결성 유용한 비 - 침습적 측정은이다. VEP는 ERG와 함께 사용될 때, 시각 시스템 (즉, 망막 / 시각 경로)의보다 완전한 평가를 제공 할 수있다. ERG 및 VEP 녹화는 애플리에 따라 단독으로 또는 조합하여 실시 할 수있다양이온. 이 문서에서 설명하는 방법은 망막과 대뇌 피질의 시각 유발 양쪽 눈에서 전기 생리학 및 마취 쥐의 두 반구의 동시 평가를 할 수 있습니다. 이것은 더 포괄적으로 망막 기능의 변화가 시각 유발 대뇌 피질의 기능에 미칠 수있는 망막 기능과 상류 효과를 평가하는 유용한 방법입니다.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<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&#246;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 ~0.7mm 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>An automatic drill for trepanning</td>
		</tr>
		<tr>
			<td>Electrode lead</td>
			<td>Grass Telefactor&#160;</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&#160;</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 watt and one 1 watt 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&#160;</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&#160;</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&#160;</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&#38;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&#160;</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 &#38; Watson&#39;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&#160;</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&#160;</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&#160;</td>
			<td>Eppendorf Pty Ltd</td>
			<td>0030 073.169</td>
			<td>Eppendorf epTIPS 100 - 5000 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.

르 A-파 (&gt; -1.38 로그 cd.sm -2), B-파도 (&gt; - 4.99 로그 cd.sm -2) STRs (&lt;- 4.99 로그 cd.sm -2)와 VEPs (&gt; - 0.52 cd.sm를 기록 -2)를 동시에 (그림 1 및 기록 하였다 3). 아주 희미한 깜박에서 긍정적 STR (pSTR)는 110 밀리 초 플래시 후 약 보지하고, 약 220 밀리 초에서 제외 STR (nSTR) (도 1 및도 2). 큰 B 파장을 가진 ERG 그 PI...

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.

전위도의 동시 녹화 및 마취 된 흰쥐의 비주얼 사건 관련 전위

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

The overall goal of this procedure is to simultaneously measure the electroretinogam and visual evoked potential in a rat. This method can help answer key questions in the field of fissure neuroscience. Such as whether the effect of aging or diseases affect certain components in the visual pathway.The main advantage of this technique is that it allows comprehensive assessment of the retinal function and its upstream affect on the visual evoked cortical function. To begin this procedure, pre-fashion the custom made ERG inactive electrodes by cutting a 70 millimeter length of silver wire and forming a loop of eight millimeters in diameter to encircle the rat eye. Prepare a uniform circle and adjust the size of the loop by shaping it on a one millimeter pipe head tip.This is important because a ring that is too small may inadvertently raise inarticular pressure, where as an oversized ring can increase the variability of signals. Next, pre-fashion the custom made VEP inactive electrode by cutting a 70 millimeter length of silver. And forming an ellipse of eight millimeters in diameter to hook on to the rat incisors.Then, pre-fashion the custom made ERG active electrodes by cutting a 30 millimeter length of silver wire and form a small loop of about one to two millimeters in diameter which is used to gently contact the rat cornea with minimal corneal abrasions. Afterward, securely attach the electrodes to the electrode leads by entwining the silver with the exposed inner wire. Insulate excess exposed metal with masking tape to reduce photo-voltaic artifacts.For the ERG active electrodes, more layers of masking tape can be applied to achieve a thicker and more rigid wrapping so that the electrode is easier to be held with a micro-manipulator. On each ERG inactive electrode, stick a small piece of hook and loop fastener to the masking tape to enable stable attachment to the rodent neck strap. Then, for each VEP active electrode, attach an alligator clip to the inner wire of the electrode lead.Immediately before recordings, electro plate the exposed surfaces of the silver wires with chloride to improve signal conduction. To do so, connect the cathode to the negative terminal of the battery and immerse the other end into the saline. Subsequently, immerse the silver tip on the anode wire into normal saline and connect the other end of this electrode wire, to the positive terminal of a nine volt battery.Disconnect them after 20 seconds and the silver tip of the electrode wire should be coated evenly in white. Repeat the chloriding procedure for all the silver ERG and VEP electrodes. In this procedure, disinfect the shaved area of an anesthetized animal with 10%povidone iodine three times.Make a sagittal incision along the mid-line on the head with a scalpel and excise a 20 millimeter diameter circle of dermal tissue to expose the cranial bone. Next, remove the underlying periosteum by scraping and drying with gauze, to expose the coronal and sagittal cranial sutures. Drill two holes through the skull on both hemispheres.Screw in stainless steel screws into the two premade holes up to a depth of one millimeter to allow firm anchorage. Then, prepare the surgical area for dental amalgam by drying the cranial bone with gauze and retracting the skin with two sutures. Subsequently, spread the dental amalgam over the exposed skull to secure the screw electrodes in place and ensure about 1.5 millimeters of the screws are exposed for recording.After that, remove the retraction sutures. For scotopic recordings, all the procedures are performed in a dark room. For illustration purposes, electrode positioning is conducted under normal room lighting here.The animal was dark adapted fore 12 hours and anesthetized with ketamine and xylazine. After pupil dilatation and topical anesthesia, hook the inactive VEP electrode around the bottom incisors. Then, place the animal on the ERG platform in front of the Ganzfeld bowl situated in the Faraday cage.Secure the animal to the platform with a strip of hook and loop fastener placed firmly but not tightly around the nape. Next, position the ERG inactive electrodes by encircling the sclaral ring non invasively around the eyes equator. Approximately three meters behind the limbus.Stabilize this by attaching the electrode to the hook and loop fastener strip around the nape. Repeat the procedure for contralateral eye. Now, fasten the VEP active electrodes by attaching alligator clips to the stainless steel screws, pre-implanted on the skull.Prior to the ERG active electrode placement, place a small drop of 1%carboxymethylcellulose sodium on the electrode to improve signal quality. Position the ERG active electrodes to lightly touch the central corneal surface using a micro manipulator attached to the custom built stereotaxic arm. Ensure the carboxymethylcellulose sodium only contacts the cornea and not the sclara, by wiping excess fluid away.Then insert two to five millimeters of the stainless steel ground needle electrode, subcutaneously into the tail. Slide the platform closer to the Ganzfeld bowl and ensure the animals eyes align with the opening of the bowl to enable even illumination of both retinas. Subsequently, close the Faraday cage to reduce extraneous noise.Using the above protocol, a stack of ERG wave forms can be recorded in response to a range of stimuli with the dimmest flash shown at the bottom and the brightest at the top. At very dim light levels, scotopic threshold responses can be recorded. With increasing stimulus brightness, the wave forms become dominated by the B wave and then the negative going A wave emerge at the brightest stimuli.The rod photoreceptor function can be assaid by using a P3 to model the A wave. This figure shows that P2 amplitude grows with increasing stimulus energy. Rod bipolar cell function can be assaid by modeling the intensity response theories of the rod P2 with a naka rushtin function.Retinal ganglion cell function is assaid at dim luminous energies and quantified by the pSTR peak amplitude and timing. Cone bipolar cell function is elicited with a paired flash paradime and quantified by cone P2 peak amplitude and timing. Amplitude analysis of the VEP is taken as peak the troph and troph to peak amplitudes which gives the timing of these responses.The amplitude of the VEP wave forms increasing with increasing stimulus energy. When it's mastered, this technique can be done within about 30 to 45 minutes excluding the time for preparing the electrode and placing the electrode and adakadaptation. While attempting this procedure, it is particularly important to pay attention to how accurately you place the electrodes and allow enough time for adakadaptation.Following this procedure, other procedures such as disease modification or therapies can be performed to answer specific questions. After watching this video, you should have a good understanding of how to prepare and place the electrodes as well as how to record and analyze the ERG and the VEP wave forms.

Research

JoVE Journal

Neuroscience

Simultaneous Electrophysiological Recording and Calcium Imaging of Suprachiasmatic Nucleus Neurons

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in ...

연구

신경과학

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

fMRI를 유도 경 두개 자기 자극하는 동안 기록 된 EEG 데이터에서 시각 유발 전위를 추출

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

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

동시<em&gt; 생체</em두 망막의&gt; 기능 테스트로<em&gt; 생체</em&gt; 전위도 시스템

An In vivo electroretinogram (ERG) signal is composed of several overlapping components originating from different retinal cell types, as well as noise ...

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

실험 시신경의 탈수 초화의 쥐 모델에서 시각 유발 전위 기록

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 ...

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

심각한<em&gt; In Vivo</em&gt; 마취 된 래트에서의 Hyperdirect 경로로부터의 국소 장 전위 및 다중 단위 활성의 전기 생리 학적 기록

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the ...

Stimulus-specific Cortical Visual Evoked Potential Morphological Patterns

자극 특이적 피질 시각적 자극 잠재적 인 형태 학적 패턴을 불러 일으켰다

This paper presents a methodology for the recording and analysis of cortical visual evoked potentials (CVEPs) in response to various visual stimuli using ...

A Method for Tracking the Time Evolution of Steady-State Evoked Potentials

Neural entrainment refers to the synchronization of neural activity to the periodicity of sensory stimuli. This synchronization defines the generation of ...

How to Obtain Reliable Visual Event-related Potentials in Newborns

신생아에서 신뢰할 수있는 시각적 이벤트 관련 잠재력을 얻는 방법

The present study discusses the characteristics of visual event-related potentials (VEPs) and outlines methodological steps for obtaining reliable ...

Objectively Assessing Sports Concussion Utilizing Visual Evoked Potentials

시각적 유발 잠재력을 활용한 스포츠 뇌진탕 객관적 평가

A portable system capable of measuring steady-state visual-evoked potentials (SSVEP) was developed to provide an objective, quantifiable method of ...

Optrode Array for Simultaneous Optogenetic Modulation and Electrical Neural Recording

동시 광유전학적 변조 및 전기 신경 기록을 위한 광섬유 어레이

During the last decade, optogenetics has become an essential tool for the investigation of neural signaling due to its unique capability of selective ...