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).

윤리 문 : 동물 실험은 과학적 목적을위한 동물의 관리 및 사용 (2013)의 호주 코드에 따라 실시 하였다. 동물 윤리 승인은 동물 윤리위원회, 멜버른 대학에서 얻었다. 재료는 본 명세서 만 아니라 의학적 또는 수 의학적 용도를위한 실험실 실험이다. 1. 준비 전극 주 : 세 채널 송신기가 동시에 수행 될 2 ERG 1 VEP 기록 가능 외과 이식에 사용된다. 활성 비활성 및 3 개의 전극을 눈에 부착하기 위해 주입 전에 링 형상으로 사전 식 될 필요가있다. 비활성 전극 전체 색깔 덮개로 덮여있는 동안 식별 목적을 위해, 제조 업체, 절반 흰색에 반 색깔의 플라스틱 덮개를 활성 전극을 동봉했다. 접지 전극 (투명 플라스틱 외피) 변경되지 않고 남아있다. 모든 활성 및 비활성 ELECTR에 대한송시 행위는 1.1, 1.2, 1.3 및 1.7 단계를 반복합니다. <ol><li> 이 뾰족한 펜치로 이중 가닥 스테인레스 스틸 전극을 꼬인 것이 풀리다. </li><li> 링 전극을 형성하는 나머지 하나의 긴 직선 가닥두고 스테인리스 스트랜드 (선단으로부터 약 1cm) 중 하나를 낸다. </li><li> 전극의 팁에서 매끄러운 고리를 형성하고, 다시 그 자체 트위스트 상 스테인리스 단일 가닥 접어. </li><li> 르 활성 전극 패션이 루프 ~ 0.2 - 루프의 염기 꼬아 직경 0.5 mm (여기에 설명 된 목적을 양쪽 눈으로부터 ERG를 기록하기 위해 이러한 방식으로 두 개의 활성 전극을 형성)을위한 ERG 비활성 VEP 전극 루프 직경은 ~ 0.8 mm 직경 (이 예에서는 하나의 능동 VEP 전극과 세 비활성 전극 용 이렇게) 만든다. </li><li> 스테인리스 나사 (직경 0.7 mm, 길이 3mm) 때문에 전극이 스크루 헤드의 주위에 얹혀 원형 VEP 활성 전극 후크. </li><li> 출동제 스테인리스 나사 (직경 0.7 mm, 길이 3 mm)의 주위에 3 비활성 전극 (ERG이 1을 VEP) K. </li><li> 자극을 줄이기 위해 두 개의 스테인리스 스틸 가닥의 날카로운 끝을 통해 앞으로 플라스틱 슬리브를 당깁니다. </li><li> 약 25 ° C에서 10 개 이상의 시간 동안 2 % 글루 타르 알데히드에 몸을 담근하여 원격 측정 송신기를 소독. 그런 다음 멸균 생리 식염수 3 회와 송신기를 씻어. </li></ol> 2. 송신기 주입 <ol><li> 동물 준비 <ol><li> 70 % 에탄올로 세정하여 실험 전에 수술 영역 소독. 사용하기 전에 모든 수술 장비를 압력솥와하지 않을 때 수술 중 사용 클로르헥시딘 장비를 유지한다. 멸균 환경을 유지하기 위해 수술 중 수술 용 드레이프 동물 커버. 모든 실험자 수술 마스크, 멸균 장갑과 가운을 착용해야합니다. </li><li> 3 L / 분 maintai의 유량으로 2 % 이소 플루 란 - 1.5 마취 유도수술을 통해 2 L / 분에서 2 % - 1.5 네드. 발가락 사이의 근육을 곤란에 페달 반사의 부재에 의해 마취의 충분한 깊이를 확인합니다. </li><li> 흉골에 사타구니 위에서 복부를 통해 40mm X 30mm의 영역을 면도. </li><li> 눈에 이마를 통해 후방을 30mm의 X 20mm의 영역을 면도하고 귀에 전방. </li><li> 두 면도 영역을 소독. 이마 지역의 경우 10 % 포비돈 - 요오드 세 번 (외과 기술자 협회에 의해 설정 연습의 표준에 부합되고, 눈 주변 지역에 대한 알코올 계 소독제의 사용을 방지)으로 소독. 복부에 걸쳐 10 %, 포비돈 요오드 및 70 % 에탄올로 소독. </li><li> 추가 국소 마취의 각막에 proxymetacaine 1 방울을 적용합니다. </li><li> 눈의 건조를 방지하기 위해 각막에 카르복시 메틸 셀룰로오스 나트륨 1 방울을 적용한다. </li></ol></li><li> 외과 이식 <ol><li> 10 밀리미터 절개를합니다수술 용 메스와 귀 사이의 수직 중간 선을 따라 머리에. </li><li> 흉골 아래의 중간 선을 따라 피부 층을 통해 복부에 5mm의 절개를합니다. </li><li> 터널 헤드 절개에 복부 절개에서 5mm 직경 캐 뉼러 피하. </li><li> 머리 복부에서 캐 뉼러를 통해 상기 송신기의 전극 배선 (3 활성 및 비활성 3)를 공급. </li><li> 송신기베이스 기준 전극을두고 무균 거즈에 전극의 끝 부분을 커버합니다. </li><li> 무균 거즈와 전극의 끝 (3 활성 및 비활성 3)을 커버. </li><li> 정위 플랫폼에 쥐의 머리를 고정합니다. </li><li> 수술 가위로 길이 30mm로 이마의 절개를 확장합니다. </li><li> 3 ~ 9시에 - 2 봉합 (0 ~ 3)으로 느슨한 피부를 수축하여 수술 영역을 노출. </li><li> 브레 그마, 람다 및 정중선 봉합을 노출 멸균 거즈를 이용하여 두개골을 덮는 골막 긁어. </리&gt; <li> VEP 활성의 두개골을 통해 두 개의 구멍을 드릴 정위 좌표를 (정중선에서 정수리 5 밀리미터 주동이) 및 비활성 (7mm은 복부 정중선 외측 3mm를 정수리합니다). </li><li> 작은 스크류 드라이버 미리 만들어진 구멍에 깊이 ~ 1mm에와 두개골에 미리 부착 된 스테인레스 스틸 나사 (직경 0.7 mm, 길이 3mm)로 VEP 활성 및 비활성 전극을 연결합니다. 이것은 기본 피질 조직을 손상시키지 않고 뼈 나사 앵커. </li><li> 일시적으로 눈꺼풀을 철회 0 봉합 - 르 활성 전극 8을 사용하여 이식합니다. </li><li> 우수한 결막 원개까지 눈 뒤에서 16 G 21 정맥의 피하에 삽입합니다. </li><li> 가이드 바늘을 제거합니다. </li><li> 눈으로 이마에서 단축 플라스틱 카테터를 통해 활성 전극을 공급. 그런 다음 플라스틱 카테터를 제거합니다. </li><li> ELEC 않도록, 전극 루프를 통해 나사 결합, - 임시 봉합사 (0-8)를 사용하여다시 터널로 후퇴에서 trode. </li><li> 12시, 윤부 뒤에 1mm에서 우수한 결막에 0.5 mm의 절개를합니다. 기본 공막을 노출 무딘 절개를 사용합니다. </li><li> 반 공막 두께의 윤부 바로 뒤에 0 봉합 - 0 또는 9 - 8을 이식. </li><li> ERG 활성 전극에서 임시 봉합을 제거합니다. </li><li> 전극의 끝 부분을 보장 3 연속 매듭을 묶는하여 반 공막 두께 봉합에 ERG 활성 전극을 고정은 윤부 부근에 자리 잡고 있습니다. </li><li> (0 - - 9 0 8) 1 ~ 2 중단 봉합사를 이용하여 결막 플랩을 닫습니다. 결막이 완전히 편안함을 개선하기 위해 ERG 전극을 커버 있는지 확인합니다. </li><li> 눈꺼풀 후퇴 봉합사를 제거합니다. </li><li> 반대편 눈의 절차를 반복합니다. </li><li> 모든 스테인리스 나사와 전극 와이어를 확보하기 위해 두개골을 통해 시아 노 아크릴 레이트 젤을 적용합니다. 확인합니다 ERG 활성 전극은 실내에 고정하기 전에 너무 꽉 끌어되지 않습니다수 안구의 움직임. </li><li> 0 봉합 - 비 흡수성 3를 사용하여 머리에 상처를 닫습니다. </li><li> 복부를 노출 설치류를 돌립니다. 수술 가위로 원격 교육 알바를 따라 40mm의 복부 피부 절개를 길게. </li><li> 내부 복강을 노출 내부 근육 벽을 통해 35 밀리미터 절개를합니다. </li><li> 이 봉합 (가) 3 - 0 ()를 사용하면 동물의 오른쪽 내부 복부 벽에 송신기 본체를 연결합니다. 간 접촉하지 마십시오. </li><li> 루프 접지 전극과 봉합사 (3-0)이 모양 보안. 이 복강에 자유 떠 놓습니다. </li><li> 연속 봉합 (0 - 3)를 사용하여 복막을 닫습니다. </li><li> 중단 봉합 (0 - 3)를 사용하여 피부 절개를 닫습니다. </li></ol></li><li> 수술 후 관리 <ol><li> 이 흉골 드러 누움을 유지하기 위해 충분한 의식을 회복 할 때까지 동물을 모니터링합니다. 집은 동물 단독으로 수술을 다음과 같습니다. </li><li> carpr 관리OFEN 피하 4 일 동안 하루에 한 번 진통제 (5 ㎎ / ㎏)합니다. </li><li> 칠일 후 수술을 위해 식수에 예방 경구 항생제 (Enrofloxin, 5 ㎎ / ㎏)를 추가합니다. </li><li> 첫 번째 7 일 후 수술 자극을 줄이기 위해 피부 절개 사이트에 항 염증 연고를 적용합니다. </li></ol></li></ol> 의식 쥐 3. 행동 ERG와 VEP 녹음 <ol><li> 다크 전에 ERG와 VEP 녹음 12 시간 동안 동물을 적용 </li><li> 어두운 붉은 조명 (17.4 cd.m -2, λ 최대 = 600 ㎚)에서 모든 실험 조작을 실시 </li><li> 국소 마취 (0.5 % proxymetacaine) 및 확장시키고 (0.5 % tropicamide) 각막에 방울을 적용합니다. </li><li> 사용자 정의 만든, 명확한 제한 수단으로 의식 쥐를 안내합니다. 주 :이 플라스틱 튜브의 길이는 60mm로 고정 전체적인 직경이 다른 크기의 래트를 수용하도록 조절 될 수있다. 상기 장치의 전방 단부는 헤드 (MO)을 최소화하기 위해 테이퍼vement과 정상적인 호흡 수 있도록 구멍이 포함되어 있습니다. 이 테이퍼 전면는 Ganzfeld 구의 개방에 정렬 및 쥐의 머리와 눈의 안정화 할 수 있습니다. 설치류가 제한 수단 (3 ~ 5 차례) 이전에 수술에 순응되어 있습니다. </li><li> 그릇의 입구에 정렬 된 눈을 가진 Ganzfeld 그릇 앞의 쥐를 놓습니다. </li><li> 송신기의 ~ 5cm 내에서 자석을 전달하여 내주 송신기의 전원을 켭니다. 송신기는 수신기베이스의 LED 상태 표시등을 확인하여 켜져 있는지 확인합니다. </li><li> 이전 21 설명 된 바와 같이 발광 에너지 (즉, -5.6 1.52 로그에 cd.sm -2)의 범위에서 신호를 수집합니다. 간략하게, 밝게 빛나는 에너지의 주차 빛 수준 (~ 80 반복) 이하에서 평균 이상의 신호 (1 ~ 반복). 점차적으로 가장 어두운 영역에서 가장 밝은 빛 레벨 1 180 초에서 interstimulus 간격을 연장. </li><li> 르로드를 분리하려면콘 응답은 트윈 플래시 패러다임 (8)을 사용한다. 예를 들어, 1.52 로그 cd.sm 두 깜박 -2 년 사이에 500 밀리 초 간 자극 간격을 시작합니다. </li><li> 밝은 발광 에너지에서 VEP 신호, 평균 20 반복을 기록하기 위해 (즉, 1.52 로그 cd.sm -2, 5 초 간 자극 간격). </li><li> ERG 및 VEP 기록 7, 10, 14, 21 및 28 일 후 수술을 수행하는 시간이 지남 신호 변동에 의해 평가된다 임플란트 안정성을 평가한다. </li><li> 자일 라진 마취 (12 : 1 ㎎ / ㎏) 실험 기간에 따라, 케타민 후 pentobarbiturate의 intracardial 주입 (1.5 ㎖ / ㎏)를 통해 쥐를 안락사. </li></ol>

<ol>
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RG and RF are employees to the commercial funder of this research (Pfizer Neusentis and Pfizer Global Research). MI was an employee of Pfizer Global Research during this research and is currently an employee of Proteostasis Therapeutics (Cambridge, USA).

때문에 시각 전기 생리학의 최소 침습 자연, 인간 환자의 ERG와 VEP 기록은 의식이있는 상태에서 수행에만 전극 배치를위한 국소 마취제의 사용을 요구하고 있습니다. 반면에, 동물 모델에서 시각적 전기 생리학 종래 자발적 눈과 몸체 움직임을 제거하여 안정적인 전극의 배치를 가능하게 전신 마취 하에서 수행된다. 이전 공보 25-27 24 등으로 도시하지만, 일반적으로 사용되는 전신...

르와 VEP는 각각 실험실과 병원 모두에서 망막과 시각 경로의 무결성을 평가하는 생체 도구의 최소 침습 있습니다. 전체 필드 ERG는 망막 통로 1,2- 다양한 세포 종류를 대표하는 각각의 요소와 다른 구성 요소들로 분해 될 수있는 특성 파형을 산출한다. 고전 전체 필드 ERG 파형은 광 수용체 활동 후 빛에 노출 2-4을 대표하는 것으로 나타났다 초기 음의 기울기 (A-파)로 구성되어 있습니다. 다음은 파가 중간 망막, 주로 ON 바이폴라 세포 5-7의 전기 활동을 반영하는 상당한 양의 파형 (B-파)로 이어진다. 또한, 하나는로드 응답 (8)로부터 콘을 분리 광량 간 자극 간격을 변화 할 수있다. 플래시 VEP는 망막 빛 자극에 반응하여 시각 피질과 뇌 줄기의 전기 전위를 나타냅니다9,10. 이 파형은 레티노-geniculo-선조체 경로 11-13 쥐 11,13 다양한 V1 라미 수행 피질 처리를 나타내는 말 성분 뉴런의 활성을 반영하는 초기 요소와, 초기 및 후기 요소로 세분화 될 수있다. 따라서 ERG와 VEP의 동시 측정은 시각 경로에 관여 구조의 포괄적 인 평가를 반환합니다. 현재 동물에서 전기 생리학을 기록하기 위해, 마취 전극 안정적인 배치를 가능하게 사용된다. 이 의식 쥐 14-16에서 ERG와 VEP을 측정하려는 시도하고 있지만, 이러한 연구는 복잡 할 수 있고, 동물의 움직임과 자연의 행동 (17)을 제한하여 동물의 스트레스로 이어질 수 유선 설정을 사용하고있다. 향상된 소형화 및 배터리 수명을 포함한 무선 기술의 최근 발전과 함께, 그것은 ERG의에 대한 원격 측정 방식을 구현하는 것이 가능하다유선 녹음 장기 생존율의 개선과 관련된 스트레스를 감소 차원 VEP 기록. 원격 측정 프로브의 완전 내재화 안정적인 주입은 만성 온도, 혈압 (18), 활동 (19)의 모니터링뿐만 아니라 뇌파 (20)에 대한 성공을 입증했다. 기술의 이러한 발전은 만성 연구를위한 플랫폼의 유틸리티를 증가, 반복성과 의식 녹음의 안정성에 도움이 될 것입니다.

<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

전체 필드 전위도 (ERG) 및 시각 유발 전위 (VEP)은 실험실과 임상 모두에서 망막과 시각 경로의 무결성을 평가하는 유용한 도구입니다. 현재 전임상 ERG와 VEP 측정은 안정된 전극 게재 위치를 보장하기 위해 마취로 수행됩니다. 그러나 마취 임재 정상적인 생리적 반응을 오염시키는 것으로 밝혀졌다. 이 마취 교란을 극복하기 위해, 우리는 의식 쥐의 ERG와 VEP를 분석 할 수있는 새로운 플랫폼을 개발한다. 전극은 수술 ERG을 분석하고 경막 VEP를 측정 할 수있는 시각 피질을 통해 할 수있는 눈에 서브 conjunctivally 주입한다. 진폭 및 감도 범위 / 타이밍 파라미터가 발광 에너지 증가에 ERG 및 VEP 양에 대해 분석된다. 르와 VEP 신호는 적어도 사주 후 수술 주입을위한 안정적이고 반복적으로 표시됩니다. 마취없이 ERG와 VEP 신호를 기록하는이 능력은 전임상들에 교란랩탑 설치는 임상 데이터에 우수한 번역을 제공해야한다.

감광체 응답은 램과 퓨의 모델에 기초하여, 각 동물에 대한 상위 2 발광 에너지 (1.20, 1.52 로그 CSM -2)에서 ERG 응답의 초기 하강 사지의 전연 지연 가우시안 피팅에 의해 분석되고 (22)는 후드와 자작 나무 (23)에 의해 공식화. 이 공식은 진폭과 민감도 매개 변수 (그림 1C 각각 1D를) 반환합니다. 쌍곡선 함수는 또한 진폭과 민감?...

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

우리는 녹음이 마취 교란없이 실시하고 있습니다 인간의 조건에 더 유사하다 눈 (전위도) 및 의식 쥐의 뇌 (시각 잠재적 유발)에서 시각 전기 생리 신호를 측정하는 수술 주입 및 녹음 과정을 보여줍니다.

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 ...

implantation-recording-wireless-electroretinogram-visual-evoked

Research

JoVE Journal

Behavior

8.6K 조회수.  University of Melbourne.  우리는 녹음이 마취 교란없이 실시하고 있습니다 인간의 조건에 더 유사하다 눈 (전위도) 및 의식 쥐의 뇌 (시각 잠재적 유발)에서 시각 전기 생리 신호를 측정하는 수술 주입 및 녹음 과정을 보여줍니다. 

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

The Successive Alleys Test of Anxiety in Mice and Rats

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. Handley, Mithani and File et al. performed the first studies on the plus-maze design we use today, and in 1987 Lister published a design for use with mice. 
Time spent on, and entries into, the open arms are an index of anxiety; the lower these indices, the more anxious the mouse is. Alternatively, a mouse that spends most of its time in the closed arms is classed as anxious. 
One of the problems of the plus-maze is that, while time spent on, and entries into, the open arms is a fairly unambiguous measure of anxiety, time in the central area is more difficult to interpret, although time spent here has been classified as &#x201C;decision making&#x201D;. In many tests central area time is a considerable part of the total test time.
Shepherd et al. produced an ingenious design to eliminate the central area, which they called the &#x201C;zero maze&#x201D;. However, although used by several groups, it has never been as widely adopted as the plus-maze. 
In the present article I describe a modification of the plus-maze design that not only eliminates the central area but also incorporates elements from other anxiety tests, such as the light-dark box and emergence tests. It is a linear series of four alleys, each having increasing anxiogenic properties. It has given similar results to the plus-maze in general. Although it may not be more sensitive than the plus-maze (more data is needed before a firm conclusion can be reached on this point), it provides a useful confirmation of plus-maze results which would be useful when, for example, only a single example of a mutant mouse was available, as, for example, in ENU-based mutagenesis programs.

The plus-maze measures anxiety-like behaviour in rodents. There are two opposite closed and two opposite open arms; anxious rodents avoid the open arms. The central area is neither completely open nor closed, so time spent here is ambiguous and difficult to interpret. Here a modification of the plus-maze protocol eliminating this area is described.

마우스 및 쥐에있는 불안의 연속 골목길 테스트

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.  ...

연구

행동분석학

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

Neuropsychological tasks used in primates to investigate mechanisms of learning and memory are typically visually guided cognitive tasks. We have developed visual cognitive tasks for rats using the Floor Projection Maze1,2&#160;that are optimized for visual abilities of rats permitting stronger comparisons of experimental findings with other species.
In order to investigate neural correlates of learning and memory, we have integrated electrophysiological recordings into fully automated cognitive tasks on the Floor Projection Maze1,2. Behavioral software interfaced with an animal tracking system allows monitoring of the animal's behavior with precise control of image presentation and reward contingencies for better trained animals. Integration with an in vivo electrophysiological recording system enables examination of behavioral correlates of neural activity at selected epochs of a given cognitive task.
We describe protocols for a model system that combines automated visual presentation of information to rodents and intracranial reward with electrophysiological approaches. Our model system offers a sophisticated set of tools as a framework for other cognitive tasks to better isolate and identify specific mechanisms contributing to particular cognitive processes.

We describe protocols for training rats for chronic electrophysiological recordings in fully automated cognitive tasks on a Floor Projection Maze.

바닥 투사 미로를 사용하여 녹음 신경 활동에 대한 자동화 된 시각인지 작업

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

Methods to Explore the Influence of Top-down Visual Processes on Motor Behavior

Kinesthetic awareness is important to successfully navigate the environment. When we interact with our daily surroundings, some aspects of movement are deliberately planned, while others spontaneously occur below conscious awareness. The deliberate component of this dichotomy has been studied extensively in several contexts, while the spontaneous component remains largely under-explored. Moreover, how perceptual processes modulate these movement classes is still unclear. In particular, a currently debated issue is whether the visuomotor system is governed by the spatial percept produced by a visual illusion or whether it is not affected by the illusion and is governed instead by the veridical percept. Bistable percepts such as 3D depth inversion illusions (DIIs) provide an excellent context to study such interactions and balance, particularly when used in combination with reach-to-grasp movements. In this study, a methodology is developed that uses a DII to clarify the role of top-down processes on motor action, particularly exploring how reaches toward a target on a DII are affected in both deliberate and spontaneous movement domains.

It is unclear how top-down signals from the ventral visual stream affect movement. We developed a paradigm to test motor behavior towards a target on a 3D depth inversion illusion. Significant differences are reported in both deliberate, goal-directed movements and automatic actions under illusory and veridical viewing conditions.

모터 동작 톱 다운 비주얼 프로세스의 영향을 탐색하는 방법

Kinesthetic awareness is important to successfully navigate the environment. When we interact with our daily surroundings, some aspects of movement are ...

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

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

수정 된 구멍 보드 - 마우스 및 쥐의 행동,인지 및 사회적 상호 작용을 측정

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

Several neuroimaging studies have suggested that the low spatial frequency content in an emotional face mainly activates the amygdala, pulvinar, and superior colliculus especially with fearful faces1-3. These regions constitute the limbic structure in non-conscious perception of emotions and modulate cortical activity either directly or indirectly2. In contrast, the conscious representation of emotions is more pronounced in the anterior cingulate, prefrontal cortex, and somatosensory cortex for directing voluntary attention to details in faces3,4. Asperger&#39;s syndrome (AS)5,6 represents an atypical mental disturbance that affects sensory, affective and communicative abilities, without interfering with normal linguistic skills and intellectual ability. Several studies have found that functional deficits in the neural circuitry important for facial emotion recognition can partly explain social communication failure in patients with AS7-9. In order to clarify the interplay between conscious and non-conscious representations of emotional faces in AS, an EEG experimental protocol is designed with two tasks involving emotionality evaluation of either photograph or line-drawing faces. A pilot study is introduced for selecting face stimuli that minimize the differences in reaction times and scores assigned to facial emotions between the pretested patients with AS and IQ/gender-matched healthy controls. Information from the pretested patients was used to develop the scoring system used for the emotionality evaluation. Research into facial emotions and visual stimuli with different spatial frequency contents has reached discrepant findings depending on the demographic characteristics of participants and task demands2. The experimental protocol is intended to clarify deficits in patients with AS in processing emotional faces when compared with healthy controls by controlling for factors unrelated to recognition of facial emotions, such as task difficulty, IQ and gender.

An EEG experimental protocol is designed to clarify the interplay between conscious and non-conscious representations of emotional faces in patients with Asperger&#39;s syndrome. The technique suggests that patients with Asperger&#39;s syndrome have deficits in non-conscious representation of emotional faces, but have comparable performance in conscious representation with healthy controls.

아스퍼거 증후군의 감정적 인 얼굴의 의식과 비 의식 표현

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

Ancient Chinese literature has documented that acupuncture possesses efficient therapeutic effects on epilepsy and insomnia. There is, however, little research to reveal the possible mechanisms behind these effects. To investigate the effect of acupuncture on epilepsy and sleep, several issues need to be addressed. The first is to identify the acupoints, which correspond between humans, rats, and mice. Furthermore, the depth of insertion of the acupuncture needle, the degree of needle twist in manual needle acupuncture, and the stimulation parameters for electroacupuncture (EA) need to be determined. To evaluate the effects of acupuncture on epilepsy and sleep, a feasible model of epilepsy in rodents is required. We administer pilocarpine into the left central nucleus of the amygdala (CeA) to simulate focal temporal lobe epilepsy (TLE) in rats. Intraperitoneal (IP) injection of pilocarpine induces generalized epilepsy and status epilepticus (SE) in rats. Five IP injections of pentylenetetrazol (PTZ) with a one-day interval between each injection successfully induces spontaneous generalized epilepsy in mice. Recordings of electrocorticograms (ECoGs), electromyograms (EMGs), brain temperature, and locomotor activity are used for sleep analysis in rats, while ECoGs, EMGs, and locomotor activity are employed for sleep analysis in mice. ECoG electrodes are implanted into the frontal, parietal, and contralateral occipital cortices, and a thermistor is implanted above the cerebral cortex by stereotactic surgery. EMG electrodes are implanted into the neck muscles, and an infrared detector determines locomotor activity. The criteria for categorizing vigilance stages, including wakefulness, rapid eye movement (REM) sleep, and non-REM (NREM) sleep are based on information from ECoGs, EMGs, brain temperature, and locomotor activity. Detailed classification criteria are stated in the text.

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

This paper demonstrates the performance of acupuncture, epilepsy models, and the analysis of sleep in rodents. The acupuncture procedure and the identification of acupoints are described. Pilocarpine or pentylenetetrazol (PTZ) is used to induce epilepsy. Electrocorticogram (ECoG), electromyogram (EMG), brain temperature, and locomotor activity recordings are employed for sleep analysis.

침술에 의해 쥐와 마우스에서 간질 Electrocorticograms (ECoGs)의 조작 및 절전

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

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer scale. However, current methods mostly rely on tethered cable recordings or only use unidirectional systems, allowing either recording or stimulation of neural activity, but not at the same time or same target. Here, a new wireless, bidirectional device for simultaneous multichannel recording and stimulation of neural activity in freely behaving rats is described. The system operates through a single portable head stage that both transmits recorded activity and can be targeted in real-time for brain stimulation using a telemetry-based multichannel software. The head stage is equipped with a preamplifier and a rechargeable battery, allowing stable long-term recordings or stimulation for up to 1 h. Importantly, the head stage is compact, weighs 12 g (including battery) and thus has minimal impact on the animal&#180;s behavioral repertoire, making the method applicable to a broad set of behavioral tasks. Moreover, the method has the major advantage that the effect of brain stimulation on neural activity and behavior can be measured simultaneously, providing a tool to assess the causal relationships between specific brain activation patterns and behavior. This feature makes the method particularly valuable for the field of deep brain stimulation, allowing precise assessment, monitoring, and adjustment of stimulation parameters during long-term behavioral experiments. The applicability of the system has been validated using the inferior colliculus as a model structure.

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

A wireless, bidirectional system for multi-channel neural recordings and stimulation in freely behaving rats is introduced. The system is light and compact, thus having minimal impact on the animal&#180;s behavioral repertoire. Moreover, this bidirectional system provides a sophisticated tool to assess causal relationships between brain activation patterns and behavior.

무선, Vivo에서 녹음 하 고 자유롭게 행동 하는 쥐에서 신경 활동의 자극에 대 한 양방향 인터페이스

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

This article explores a method to detect differences in visual perception in humans. The method used is based on the psychological (or "cognitive") function of eyeblinks. Participants' eyeblinks are detected and acquired while watching videos specifically created for the investigation. The detection and acquisition of eyeblinks are carried out with the help of a 20-channel electroencephalographic (EEG) wireless device. The international 10-20 system for electrode placement is followed. A high-definition (HD) video camera is used to record participants' facial expressions, for contrast purposes. Instead of using pre-existing media content, purpose-made video content has been created following specific criteria of interest for this investigation, with stimuli enabling researchers to manage the precise parameters of interest. Otherwise, results could be contaminated with uncontrolled variables. The synchronization of the presentation of video stimuli with EEG recordings needs to be done in milliseconds. Analysis of collected data is performed with robust software for working with big matrices. Statistically significant differences in eyeblink rate related to media professionalization and editing style are found with the reported experimental procedures.

We present detection, acquisition, and analysis of eyeblink rates while watching media content.

Electroencephalography 측정 및 높은-품질 비디오 녹화를 사용 하 여 미디어 콘텐츠의 시각적 인식 분석

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

The present work is a description and an assessment of a methodology designed to quantify different aspects of the interaction between language processing and the perception of the visual world. The recording of eye-gaze patterns has provided good evidence for the contribution of both the visual context and linguistic/world knowledge to language comprehension. Initial research assessed object-context effects to test&#160;theories of modularity in language processing. In the introduction, we describe how subsequent investigations have taken the role of the wider visual context in language processing as a research topic in its own right, asking questions such as how our visual perception of events and of speakers contributes to&#160;comprehension informed by&#160;comprehenders&#39; experience. Among the examined aspects of the visual context are actions, events, a speaker&#39;s gaze, and emotional facial expressions, as well as spatial object configurations. Following an overview of the eye-tracking method and its different applications, we list the key steps of the methodology in the protocol, illustrating how to successfully use it to study visually-situated language comprehension. A final section presents three sets of representative results and illustrates the benefits and limitations of eye tracking for investigating the interplay between the perception of the visual world and language comprehension.

The present article reviews an eye-tracking methodology for studies on language comprehension. To obtain reliable data, key steps of the protocol must be followed. Among these are the correct set-up of the eye tracker (e.g., ensuring good quality of the eye and head images) and accurate calibration.

시각적으로 위치한 언어 이해 동안 눈 추적: 유연성과 잠복 시각적 컨텍스트 효과에 한계

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

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, studies implementing classical conditioning paradigms with harnessed insects have been important for investigating the exact cues that individuals learn and the neural mechanisms underlying learning and memory formation. Here we present a protocol for evoking visual associative learning in wood ants through classical conditioning. In this paradigm, ants are harnessed and presented with a visual cue (a blue cardboard), the conditional stimulus (CS), paired with an appetitive sugar reward, the unconditional stimulus (US). Ants perform a Maxilla-Labium Extension Reflex (MaLER), the unconditional response (UR), which can be used as a readout for learning. Training consists of 10 trials, separated by a 5-minute intertrial interval (ITI). Ants are also tested for memory retention 10 minutes or 1 hour after training. This protocol has the potential to allow researchers to analyze, in a precise and controlled manner, the details of visual memory formation and the neural basis of learning and memory formation in wood ants.

We present a protocol for the classical conditioning of harnessed ants that permits researchers to study visual learning and memory formation at a level of analysis not possible with freely moving individuals.

나무 개미에 비주얼 고전 컨디셔닝

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