Name: visual evoked potential recording in a rat model of experimental optic nerve demyelination
Uploaded: 2015-07-29T14:00:00.000+00:00
Duration: 6 min 49 s
Description: Watch this Scientific Journal Video about Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination at JoVE.com

이 연구는 호주 안과 연구소 (오리 아)에 의해 지원되었다. 우리는 처음에 VEP 기록 기술을 개발하기 위해 우리를 도와, 교수 Algis Vingrys 박사 빅뱅 부이, 멜버른 대학 감사합니다.

윤리 문 : 동물과 관련된 모든 절차는 관리 및 과학적인 목적을위한 동물의 사용 및 안과 및 비전 연구에서 동물의 사용에 대한 ARVO 방침의 지침 연습 호주 코드에 따라 수행되었으며, 승인했다 맥쿼리 대학의 동물 윤리위원회. 1. VEP 전극 주입 <ol><li> 복강 내 주사 케타민 (75 ㎎ / ㎏) 및 메데 토미 (0.5 밀리그램 / kg)으로 동물을 마취. 마취의 유도에 따라 수술을 시작하는 표시로 철수 반사 (핀치 검사, 각막과 눈꺼풀 반사 등)과 부재을 준수합니다. 수술 내내 계속 동물을 모니터링하고 반사가 존재하는 경우 추가 마취약 (상위 - 업 케타민 당 초기 용량의 10 %)을 관리. 성인 쥐 (&gt; 12주)는 실험에 사용된다. </li><li> 외과 영역의 피부를 면도.수술 중 체온을 유지하기 위해 온난화 패드 (37 ° C)에서 동물을 놓습니다. 포비돈 - 요오드의 국소 응용 프로그램을 통해 피부를 준비합니다. 수술 입체 재단을 적용합니다. 전신 마취하에 각막 건조를 방지하기 위해 국소 안과 연고를 적용합니다. 멸균 악기를 사용하여 무균을 유지한다. </li><li> 헤드 피부의 중간 선에 길이의 피부 절개를합니다. 두개골의 좋은 노출을 달성하기 위해 결합 조직을 취소합니다. </li><li> 조심스럽게, 수동으로 브레 그마 뒤에 7mm과 중간 선에 3mm 측면에서 마이크로 핸드 드릴을 사용하여 작은 버 구멍을 드릴. </li><li> 약 0.5 mm의 깊이로 피질을 관통하는 피질 (영역 (17))에 두개골을 통해 임플란트 나사 전극. 브레 그마로 정중선 3mm의 입쪽에 기준 나사 전극을 이식. 나사를 싸는하고 해결하기 위해 치과 시멘트를 적용합니다 (항상 필요하지 않음). </li><li> RECO하는 동물을 머리의 피부 봉합 피부에 항생제 연고를 관리 허용버전 온난화 패드에 마취에서. 주 : 대안 적으로, 전극이 노출 남아있을 수 있다는 것을 피부에서 각각의 기록 재개 할 필요가없는, 그래서. 수술 전에 즉시하지만 마취 회복의 정지시에, 비 스테로이드 성 항 염증 약물 (NSAID) (아니라면 사전에 동작 투여) 또는 아편 유사 진통제를 투여. 마취 완전히 외래에서 완전히 회복 될 때까지 지속적으로 동물을 모니터링합니다. </li><li> 동물이 수술의 사전 VEP 기록을 복구하는 적어도 일주 허용합니다. </li></ol> 2. 시신경 주입 <ol><li> , 동물을 마취 피부를 준비하고 (1.1 및 1.2) 이상으로 드레이프 적용됩니다. </li><li> 무작위로 선택된 눈의 궤도 위의 피부에 1 - 1.5 cm의 절개를합니다. 미세 홍채 가위를 사용하여 궤도 캐비티에 도달하기 위해 피하 조직을 엽니 다. 결막을 열고 운영 현미경으로 장부의 캡슐을 전방. </li><li> 안구 후퇴근육과 intraobital 눈물 분비는 시신경의 약 3mm 길이를 노출합니다. 길이 방향으로 안과 블레이드를 사용하여 시신경 주위의 경막과 지주막 물질 층을 엽니 다. </li><li> 세계에 2mm 후방의 거리에서 시신경에 유리 피펫을 삽입합니다. 유리 마이크로 피펫은 해밀턴 주사기에 부착된다. </li><li> 약 30 초에 걸쳐 천천히 신경으로 - (수초에 효과가없는 0.02 % 에반 블루와, 1.0 μL, 0.4), 1 % 리소 레시틴을 주입한다. </li><li> 피부 절개 봉합. 감염을 방지하기 위해 항생제 연고를 적용합니다. 동료의 눈은 전기 생리학 레코딩을위한 내부 통제 역할을 할 수있다. </li><li> 마취에서 회복 온난화 패드에 동물을 놓습니다. </li></ol> 3. VEP 기록 <ol><li> 동물을 마취하고 1.1 및 1.2로 피부를 준비합니다. 참고 : 마취제의 낮은 복용량은 전기 생리 RECO 사용할 수 있습니다rding (케타민 40 ㎎ / ㎏과 0.25 ㎎ / ㎏을 메데 토미). </li><li> 어두운 방에서 쥐를 놓고는 5 어둠에 적응 할 수 - 30 분. 어떤 경우에는 쥐가 각각 포토 픽 VEP 녹음 8 적응 암순응 또는 빛 어두운 적응 O / N이 될 수 있습니다. </li><li> 직장 온도계 프로브 homoeothermic 블랭킷 시스템에 의해 37 ± 0.5 ℃의 체온을 유지한다. </li><li> 1.0 % tropicamide 눈 방울과 학생들을 팽창. 액세스하기 위해 두개골을 통해 피부를 열고 사전 배치 현장 나사 전극에서. </li><li> 자극 눈의 반대편 시각 피질과 앰프에 대한 참조 나사를 통해 나사를 연결합니다. 지상으로 꼬리에 바늘 전극을 삽입합니다. 측정하고 5 kΩ의 아래 전극 임피던스를 유지한다. </li><li> 뛰어난 눈 절연 (7)을 제공하기 위해 눈꺼풀 주위의 피부에 직접 미니 Ganzfeld 자극을 놓습니다. 자극기의 조명이 미리 보정해야광도계에 의해. </li><li> 각각 1 내지 100 Hz의 낮은 및 높은 밴드 패스 필터 설정을 1 Hz의 주파수에서 빛을 깜박 100 회 빛의 자극을 전달한다. 신호 샘플링 레이트는 5 kHz에서이다. 참고 : 신호는 적어도 약 250 샘플링한다 - 더 두 개의 샘플은 각주기 동안 수집되는 것을 보장하기 위해 300 Hz에서. </li><li> 다시 피부를 봉합하고 마취에서 회복하기 온난화 패드에 동물을 유지. 녹화 반복 시간 기간 동안에 기능적 변화를 모니터링하기 위해 각각의 동물에 기록 될 수있다. </li><li> 최종 시점에서, 동물을 안락사 나트륨 펜토 바르비 톤 (100 밀리그램 / kg, IP)의 과다 주사를 관리. 심장 마비, 호흡 정지 및 체온의 감소에 의해 안락사를 확인합니다. </li></ol> 4. 조직 준비 및 조직학 <ol><li> 현미경으로 안락사 동물에서 시신경을 제거하고 1 % 파라 포름 알데히드의 O / N의 조직을 고정합니다. </li> &lt;리&gt; 식염수로 철저하게 조직을 씻으십시오. 자동 조직 프로세서에서 처리하고, 조직을 파라핀에 포함. 회전식 마이크로톰을 사용하여 - (10 μm의 5) 컷 섹션. 참고 : 면역 조직 화학 연구를 들어, 1 % 파라 포름 알데히드에 조직을 고정 식염수로 세척하고 30 % 자당 O / N로 품어. OCT에서 임베드 조직 매체를 포함하고 저온 ​​유지 장치를 사용하여 저온부을합니다. </li><li> 56 ° C에서 같은 룩솔 (95 % 에탄올) O / N으로 0.1 % 빠른 블루 솔루션 섹션을 품어. 다음 다른 30 ​​초 70 % 에탄올을 30 초 동안 0.05 %의 탄산 리튬의 섹션을 차별화하고. 마지막 섹션을 장착하기 전에 30 초 동안 0.1 % 크레 실 바이올렛 용액 Counterstain과. 시신경 5 수초를 식별하기 위해 빠른 파란색 염색을 사용합니다. </li></ol>

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None of the authors have competing interests or conflicting interests.

The optic nerve is very susceptible to mechanical damage. Optic nerve crush injury over a duration of 1 s can lead to about 75% loss of RGC over a period of 2 weeks10. Therefore, extreme care is required while performing the surgical procedures. According to the authors&#8217; experience, it is much better to adapt a blunt dissection approach to expose and make way through the tissues around the optic nerve along the orientation of the nerve, rather than penetrating in a perpendicular orientation to the optic ...

Optic neuritis is one of the most common form of optic neuropathy, causing complete or partial loss of vision1. Histologically, it is featured by inflammatory demyelination, retinal ganglion cell axonal loss and varying degrees of remyelination in the optic nerve2. Optic neuritis is usually the manifest onset of multiple sclerosis. The visual evoked potential (VEP) is a non-invasive tool for investigating the function of the visual system. It reflects the post-retinal function from the retina to the primary visual cortex and is affected in many optic nerve disease conditions3. The VEP has been predominantly used in optic neuritis patients to assess the integrity of the visual pathway4. 
The latency of VEP, which reflects the velocity of signal conduction along the visual pathway, is considered to be an accurate measurement of the level of myelin associated changes in the optic nerve5; while the amplitude of VEP is believed to be closely correlated with axonal damage of the retinal ganglion cells (RGC)6. This hypothesis has been fairly well established using the rat model of lysolecithin-induced optic nerve demyelination5. 
Here, we explicate a comprehensive protocol of optic nerve microinjection technique in rodents, which can minimise the surgical manipulation-related damage to the nerve per se as well as to the adjacent tissues such as extraocular muscles and blood vessels. Also, the skull electrode implantation surgery has been described for VEP recording in animals7. The VEP recordings can be repeatedly carried out on animals over a period of time to assess demyelination/remyelination related changes as well as impact on axonal integrity in the optic nerve.

<table><tbody><tr><td>Ketamine 100 mg/ml (Ketamil)</td><td>Troy Laboratories</td><td>AC 116</td><td></td></tr><tr><td>Medetomidine 1 mg/ml (Domitor)</td><td>Pfizer</td><td>sc-204073</td><td></td></tr><tr><td>Tropicamide 1.0% (Mydriacyl)</td><td>Alcon</td><td>sc-202371</td><td></td></tr><tr><td>Homoeothermic blanket system</td><td>Harvard Apparatus</td><td>NC9203819</td><td></td></tr><tr><td>Impedance meter&amp;nbsp;</td><td>Grass</td><td>F-EZM5</td><td></td></tr><tr><td>Screw electrodes&amp;nbsp;</td><td>Micro Fasteners</td><td>M1.0&amp;times;3mm Csk Slot M/T 304 S/S</td><td></td></tr><tr><td>Subdermal needle electrodes&amp;nbsp;</td><td>Grass</td><td>F-E3M-72</td><td></td></tr><tr><td>Rapid Repair&amp;nbsp;</td><td>DeguDent GmbH</td><td></td><td></td></tr><tr><td>Light-emitting diode&amp;nbsp;</td><td>Nichia</td><td>NSPG300A</td><td></td></tr><tr><td>Bioamplifier</td><td>CWE, Inc.</td><td>BMA-400</td><td></td></tr><tr><td>CED system</td><td>Cambridge Electronic Design, Ltd.</td><td>Power1401</td><td></td></tr><tr><td>Hamilton syringe&amp;nbsp;</td><td>Hamilton</td><td>87930</td><td></td></tr><tr><td>Lysolecithin</td><td>Sigma</td><td>L4129</td><td></td></tr><tr><td>Evan&amp;rsquo;s blue&amp;nbsp;</td><td>Sigma</td><td>E2129</td><td></td></tr></tbody></table>

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.

재현성 인트라 셔널 VEP 트레이스는도 1에 도시되어 있으며 대기 N1에서 상당한 지연은 시신경 주입 후에 알 수있다. 탈수 초화의 부분 시신경 병변 룩솔 패스트 블루 염색법을 이용하여 조직 학적 5 절에서 발견 될 수있다. 2 시신경의 중심에 작은 초점 탈수 초 병변 대표적인 부분을 도시한다. 단면은 병변의 총 부피를 대표하지 않습니다. 탈?...

Watch this Scientific Journal Video about Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination at JoVE.com

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

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.

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 ...

visual-evoked-potential-recording-rat-model-experimental-optic-nerve

Research

JoVE Journal

Neuroscience

12.2K 조회수.  Macquarie University. 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. 

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

Single-unit In vivo Recordings from the Optic Chiasm of Rat

Information about the visual world is transmitted to the brain in sequences of action potentials in retinal ganglion cell axons that make up the optic nerve. In vivo recordings of ganglion cell spike trains in several animal models have revealed much of what is known about how the early visual system processes and encodes visual information. However, such recordings have been rare in one of the most common animal models, the rat, possibly owing to difficulty in detecting spikes fired by small diameter axons. The many retinal disease models involving rats motivate a need for characterizing the functional properties of ganglion cells without disturbing the eye, as with intraocular or in vitro recordings. Here, we demonstrate a method for recording ganglion cell spike trains from the optic chiasm of the anesthetized rat. We first show how to fabricate tungsten-in-glass electrodes that can pick up electrical activity from single ganglion cell axons in rat. The electrodes outperform all commercial ones that we have tried. We then illustrate our custom-designed stereotaxic system for in vivo visual neurophysiology experiments and our procedures for animal preparation and reliable and stable electrode placement in the optic chiasm.

Single-unit In vivo Recordings from the Optic Chiasm of Rat

Retinal ganglion cells transmit visual information from the eye to the brain with sequences of action potentials. Here, we demonstrate how to record the action potentials of single ganglion cells in vivo from anesthetized rats.

단일 유닛 생체내에서</em암흑의 광학 Chiasm에서> 레코딩

Information about the visual world is transmitted to the brain in sequences of action potentials in retinal ganglion cell axons that make up the optic ...

연구

신경과학

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.

의식 쥐에 주입 및 무선 막전위의 기록 및 Visual 사건 관련 전위

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

행동분석학

Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history have&#160;remained poorly understood. Recently, we developed a reliable, reproducible animal model of PION and tested the treatment effect of some neurotrophic factors in this model1. The purpose of this video is to demonstrate our photochemically induced model of posterior ischemic optic neuropathy, and to evaluate its effects with retrograde labeling of retinal ganglion cells. Following surgical exposure of the posterior optic nerve, a photosensitizing dye, erythrosin B, is intravenously injected and a laser beam is focused onto the optic nerve surface. Photochemical interaction of erythrosin B and the laser during irradiation damages the vascular endothelium, prompting microvascular occlusion mediated by platelet thrombosis and edematous compression. The resulting ischemic injury yields a gradual but pronounced retinal ganglion cell dieback, owing to a loss of axonal input - a remote, injury-induced and clinically relevant outcome. Thus, this model provides a novel platform to study the pathophysiologic course of PION,&#160;and can be further optimized for testing therapeutic approaches for optic neuropathies as well as other CNS ischemic diseases.

The goal of this protocol is to photochemically induce ischemic injury to the posterior optic nerve in rat. This model is critical to studies of the pathophysiology of posterior ischemic optic neuropathy, and therapeutic approaches for this and other optic neuropathies, as well as of other CNS ischemic diseases.

광 화학적으로 유도 된 후방 허혈성 시신경 병증의 쥐 모델

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history ...

의학

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.

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 ...

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

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

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

드라이 비-침략 적 멀티 채널을 사용 하 여 마우스에 비주얼 갖는 잠재적인 녹음 두 피 뇌 파 센서

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

In Vivo Methods to Assess Retinal Ganglion Cell and Optic Nerve Function and Structure in Large Animals

The optic nerve collects axons signals from the retinal ganglion cells and transmits visual signal to the brain. Large animal models of optic nerve injury are essential for translating novel therapeutic strategies from rodent models to clinical application due to their closer similarities to humans in size and anatomy. Here we describe some in vivo methods to evaluate the function and structure of the retinal ganglion cells (RGCs) and optic nerve (ON) in large animals, including visual evoked potential (VEP), pattern electroretinogram (PERG) and optical coherence tomography (OCT). Both goat and non-human primate were employed in this study. By presenting these in vivo methods step by step, we hope to increase experimental reproducibility among different labs and facilitate the usage of large animal models of optic neuropathies.

In Vivo Methods to Assess Retinal Ganglion Cell and Optic Nerve Function and Structure in Large Animals

Here we demostrate several in vivo tests (flash visual evoked potential, pattern electroretinogram and optic coherence tomography) in goat and rhesus macaque to understand the structure and function of the optic nerve and its neurons.

인 비보 큰 동물의 망막 신경 세포 및 시신경 기능 및 구조를 평가하는 방법

The optic nerve collects axons signals from the retinal ganglion cells and transmits visual signal to the brain. Large animal models of optic nerve injury ...

Inducing Local Demyelination in the Optic Nerve Using Lysolecithin

Source: You, Y. et al., <a href="https://app.jove.com/v/52934">Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination.</a> J. Vis. Exp. (2015)
This video demonstrates the procedure of surgically accessing the optic nerve in an anesthetized animal and precisely delivering lysolecithin to induce focal demyelination, creating neuropathological conditions for further study.

리솔레시틴(Lysolecithin)을 이용한 시신경의 국소 탈수초화(local demyelination) 유도

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

JoVE Encyclopedia of Experiments

Neuropathology

Epilepsy and Seizure Disorders

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/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.

전기생리학 설정을 사용하여 시각적 경로 무결성을 평가하기 위한 RAT 모델 준비

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

Neurophysiology

Electrophysiology Techniques

Electron Microscopic Analysis of Myelin Damage in a Rat Model of Optic Nerve Injury

Take fixed optic nerve sections from a control rat and a rat with optic nerve injury.
The control section exhibits axons with tightly packed myelin sheaths containing concentric plasma membrane layers. The injured section exhibits decompacted myelin with large gaps.
Incubate with an osmium-ferrocyanide complex that binds to lipids and enhances the myelin sheath&#39;s contrast.
Treat with thiocarbohydrazide to create additional binding sites for osmium.
Reapply osmium to further enhance lipid contrast.
Stain with heavy metals, which bind to macromolecules and enhance structural contrast.
Dehydrate using increasing alcohol concentrations and rinse with acetone for resin embedding.
Embed the tissue in resin, and coat the resin with gold to improve image quality.
Use serial block-face scanning electron microscopy to simultaneously slice and image. Focus the electron beam on the surface and detect the backscattered electrons to generate images.
Myelin sheaths appear dark, while spaces within damaged sheaths appear lighter, allowing differentiation between normal and damaged myelin.

시신경 손상의 쥐 모델에서 수초(Myelin) 손상의 전자 현미경 분석(Electron Microscopic Analysis of Myelin Damage in a Rat Model of Optic Nerve Injury)

Neuromuscular Disorders

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

Source: You, Y., et al. <a href="https://app.jove.com/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.

쥐 모델의 두개골 전극을 통한 시각적 유발 전위 기록

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