이 연구는 중국 자연 과학 재단 (31171355)과 광동 자연 과학 재단 (S2011010003403, 2014A030313440)의 지원을 받았다.

동물 보호, 수술 절차 및 기록 절차는 남 중국 사범 대학 동물 관리 및 사용위원회의 지침에 따랐습니다. 1. 전극 준비 (그림 1A-C) 참고 : 컴퓨터 핀 위치는 통신 장치의 신호 인터페이스의 일부로 뾰족한 접촉입니다. 암 커넥터에 플러그되는 수 커넥터가 구성됩니다. <ol><li> 조임 장치를 사용하여 남성 핀과 여성 핀을 컴퓨터 핀 위치 ( 그림 1A )에서 조심스럽게 분리합니다. 수컷과 암컷 핀을 함께 연결하여 전극을 만들고 시아 노 아크릴 레이트를 사용하여 강한 접착 본드를 만듭니다 ( 그림 1C ). </li><li> 증류수가 들어있는 비이커에 전극을 넣고 10 분 동안 초음파 세척기에 놓습니다. 45 ° C에서 30 분 동안 건조 오븐으로 옮깁니다. 30 분 동안 자외선을 사용하여 전극을 소독하십시오. </li></ol> 2. 수술 절차 (그림 1D - F) <ol><li> 멸균 수술 도구 및 stereotaxic기구를 준비합니다. 공기와 함께 isoflurane 마취 (2.5 %)를 사용하여 신생아 쥐 강아지를 마취. 강아지가 깊이 anesthetized 때 1.0 %로 isoflurane의 복용량을 조정합니다. 적절한 마취 깊이를 보장하기 위해 수술 전에 꼬리 또는 발끝을 꼬집어 라. </li><li> 정강이 장치의 귀 머리를 귀 운하에 넣고 약간 조입니다. 참고 : 신생아 두개골이 매우 부드럽기 때문에 귀 막대를 과도하게 조이지 마십시오. </li><li> 모든 장비에 70 % 에탄올을 뿌려 멸균 수술장을 유지하십시오. 메스를 사용하여 머리에 15mm 절개를하십시오. 집게를 사용하여 네 모퉁이의 정중선에서 두피를 조심스럽게 당겨냅니다. 절개를 활짝 열어두기 위해 피부 아래에 생리 식염수를 뿌려주십시오 ( 그림 1D ). </li><li> 두개골에 bregma와 lambda 점을 찾아라.그들을 연필로 표시하십시오. 주사기 바늘 (26G)을 사용하여 전두엽 피질 (PFC)과 해마에 두 개의 구멍을 뚫습니다. 참고 : Preg는 bregma의 후방에서 + 1.8mm, 정중선에서 -0.5mm 떨어져 있으며 해마는 bregma의 앞쪽에서 -2.0mm, 정중선에서 +/- 0.5mm 떨어져 있습니다 ( 그림 1D 및 E ). 전극의 깊이는 뇌 손상을 최소화하기 위해 피질 표면 아래 2mm 이상 떨어져서는 안됩니다. </li><li> 집게를 사용하여 전극을 잡고 기준 전극과 기록 전극을 각각 PFC와 해마에 삽입하십시오. 가능한 감염을 피하기 위해 전극 주위에 에리스로 마이신 연고를 바르십시오. 시아 노 아크릴 레이트를 사용하여 전극을 고정하십시오. </li><li> 접착 성 및 점성이있는 일관성을 유지하도록 치과 용 아크릴 시멘트를 준비하십시오. 전극과 두개골의 나머지 부분을 덮기 위해 치과 용 시멘트를 바릅니다. 참고 : 치과 용 시멘트를 바르기 전에 두개골을 완전히 말립니다. </li&gt; <li> 전극을 보호하기 위해 5 % 피크 글루 산을 전극에 바릅니다. 참고 : 전체 절차는 멸균 상태를 유지하기 위해 바이오 안전성 후드에서 수행되어야합니다. </li><li> stereotaxic 프레임에서 동물을 제거하고 10 % 포도당 300 μL를 피하 주사. 가열 된 담요 위에 올려 놓으십시오. 동물이 따뜻하고 (37 ° C), 이동성이 있는지 ( 즉 완전히 회복되었는지) 확인하십시오. 수술 후 통증에 대해 부 프레 노르 핀 (0.05 mg / kg)을 복강 내 투여. 참고 : 충분한 의식 (정상적인 행동 및 움직임)이 회복 될 때까지 동물을 방치하지 마십시오. </li><li> 의식을 되찾은 후에는 댐을 가지고 강아지를 집으로 되돌려 보내십시오. 동물이 완전히 회복 될 때까지 이틀간 기다리십시오. </li></ol> 3. 뇌파 기록 <ol><li> 완전 회복 후 강아지의 두개골에 이식 된 전극을 자체 케이지에있는 증폭기에 연결하십시오. 앰프를 아날로그 - 디지털 c에 연결하십시오.컨버터를 컴퓨터에 연결하십시오. 연결 라인은 얽히게하지 않도록 조심스럽게 다루어야합니다. </li><li> 녹음을 위해 데이터 수집 장치에서 최소 10,000 Hz 샘플링 속도를 선택하십시오 (송신기의 대역폭은 1 - 100 Hz입니다). 데이터가 올바르게 샘플링되었는지 확인하십시오. </li><li> 기준 기록을 얻은 후, 간질 발작을 유도하기 위해 카 이닌산 (KA) (2 mg / kg)을 복강 내에 복강 내 주사한다. KA 주입 후 15 분 동안 간질 방출을 관찰하고 기록하십시오. KA를 통한 경련 생산은 대개 물리적 인 것입니다. 참고 : 발작 발기 지속 시간은 약 15.2 ± 0.9 초, 발작 지속 시간은 약 62 ± 5 초입니다. 염소 수화물 (400 mg / kg)을 IP 주사하여 신생아 쥐에서 발작을 예방할 수 있습니다. </li><li> 디지털화 된 데이터를 저장하고 spike2와 같은 신호 처리 소프트웨어 패키지를 사용하여 분석하십시오. perf에 의해 신생아 EEG 신호의 여러 주파수 구성 요소의 출력 수준을 보여줍니다.파워 스펙트럼 분석을 오밍하는 것. 1에서 100 Hz (EEG 밴드)까지의 Root-Mean-Square 진폭을 찾아서 1 분 시간대의 전력을 계산하십시오. </li></ol>

<ol>
	<li>Pereda, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+synapses+and+their+functional+interactions+with+chemical+synapses.">Electrical synapses and their functional interactions with chemical synapses.</a> Nat Rev Neurosci. 15 (4), 250-263 (2014).</li><li>Chorev, E., Epsztein, J., Houweling, A. R., Lee, A. K., Brecht, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+recordings+from+behaving+animals--going+beyond+spikes.">Electrophysiological recordings from behaving animals--going beyond spikes.</a> Curr Opin Neurobiol. 19 (5), 513-519 (2009).</li><li>Freeborn, D. L., McDaniel, K. L., Moser, V. C., Herr, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+electroencephalography+(EEG)+to+assess+CNS+changes+produced+by+pesticides+with+different+modes+of+action:+effects+of+permethrin,+deltamethrin,+fipronil,+imidacloprid,+carbaryl,+and+triadimefon.">Use of electroencephalography (EEG) to assess CNS changes produced by pesticides with different modes of action: effects of permethrin, deltamethrin, fipronil, imidacloprid, carbaryl, and triadimefon.</a> Toxicol Appl Pharmacol. 282 (2), 184-194 (2015).</li><li>Werhahn, K. J., Hartl, E., Hamann, K., Breimhorst, M., Noachtar, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Latency+of+interictal+epileptiform+discharges+in+long-term+EEG+recordings+in+epilepsy+patients.">Latency of interictal epileptiform discharges in long-term EEG recordings in epilepsy patients.</a> Seizure. 29, 20-25 (2015).</li><li>Staba, R. J., Stead, M., Worrell, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+biomarkers+of+epilepsy.">Electrophysiological biomarkers of epilepsy.</a> Neurotherapeutics. 11 (2), 334-346 (2014).</li><li>Nimmrich, V., Draguhn, A., Axmacher, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+Network+Oscillations+in+Neurodegenerative+Diseases.">Neuronal Network Oscillations in Neurodegenerative Diseases.</a> Neuromolecular Med. 17 (3), 270-284 (2015).</li><li>Zayachkivsky, A., Lehmkuhle, M. J., Dudek, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+Continuous+EEG+Monitoring+in+Small+Rodent+Models+of+Human+Disease+Using+the+Epoch+Wireless+Transmitter+System.">Long-term Continuous EEG Monitoring in Small Rodent Models of Human Disease Using the Epoch Wireless Transmitter System.</a> J Vis Exp. (101), e52554 (2015).</li><li>Zayachkivsky, A., Lehmkuhle, M. J., Fisher, J. H., Ekstrand, J. J., Dudek, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recording+EEG+in+immature+rats+with+a+novel+miniature+telemetry+system.">Recording EEG in immature rats with a novel miniature telemetry system.</a> J Neurophysiol. 109 (3), 900-911 (2013).</li><li>Dzhala, V. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NKCC1+transporter+facilitates+seizures+in+the+developing+brain.">NKCC1 transporter facilitates seizures in the developing brain.</a> Nat Med. 11 (11), 1205-1213 (2005).</li><li>Tucker, A. M., Aquilina, K., Chakkarapani, E., Hobbs, C. E., Thoresen, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+amplitude-integrated+electroencephalography+and+interburst+interval+in+the+rat.">Development of amplitude-integrated electroencephalography and interburst interval in the rat.</a> Pediatr Res. 65 (1), 62-66 (2009).</li><li>Savard, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Involvement+of+neuronal+IL-1beta+in+acquired+brain+lesions+in+a+rat+model+of+neonatal+encephalopathy.">Involvement of neuronal IL-1beta in acquired brain lesions in a rat model of neonatal encephalopathy.</a> J Neuroinflammation. 10, 110 (2013).</li><li>Cuaycong, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+approach+to+the+study+of+hypoxia-ischemia-induced+clinical+and+subclinical+seizures+in+the+neonatal+rat.">A novel approach to the study of hypoxia-ischemia-induced clinical and subclinical seizures in the neonatal rat.</a> Dev Neurosci. 33 (3-4), 241-250 (2011).</li></ol>

저자는 아무런 이해 상충을 선언하지 않습니다.

여기서는 자유롭게 움직이는 신생아 생쥐의 EEG를 유선 방법으로 획득하기위한 수술 및 기록 절차를보고합니다 ( 그림 3 ). P7-P12 쥐 강아지는 만삭 인간 신생아 10 , 11 에 해당하는 발육기 연령에 있다고 제안되었습니다. 이 연령 그룹의 쥐 새끼들과 작업 할 때 고품질의 뇌파 기록 데이터를 얻는 것은 기술적으로 어렵습니다. 또한 특별한 취?...

정상적인 뇌 기능을 얻기 위해서는 뉴런 간의 지속적인 통신이 필요하다는 것이 잘 알려져 있습니다. interneuronal 의사 소통은 주로 하나의 뉴런에서 정보가 두 번째 뉴런으로 전달되는 시냅스에서 발생합니다. 이 시냅스 전달은 전기적 또는 화학적 시냅스의 두 가지 유형의 전용 구조 배열에 의해 조정됩니다. 전기 생리학 (Electrophysiology)은 전반적인 신체 기능과 행동을 제어하는 ​​간세포 통신 과정에서 발생하는 전위를 포착하는 분야입니다. EEG는 많은 전기 생리 학적 기법 중에서 가장 일반적으로 사용되는 방법입니다. EEG는 내부 또는 외부 자극에 의해 생성 된 전기 신호의 변화를 감지하는 데 사용되는 기술입니다. 또한, 그것은 간질, 파킨슨 병 및 알츠하이머 병과 같은 다양한 신경 학적 증상의 임상 적 진단 및 결과 예측에 필수적인 검사입니다e뿐만 아니라 약리학 적 및 독성 학적 인자의 ​​영향 3 . 일반적으로 간질 환자는 과다 흥분성과 뇌의 기능적 연결성을 손상시킵니다. 이들은 간질 간질 방전 (IEDs)으로 요약되며 예리하고 일시적인 스파이크의 형태로 EEG로 기록 될 수 있습니다. 날카로운 파도; 스파이크 - 웨이브 복합체; 또는 polyspikes 4 . 간질 뇌의 주요 특징은 발작을 일으키는 뇌 영역을 찾기 위해 두피 또는 뇌 실질에서 기록 할 수있는 간질 발작의 자연 발생입니다. 또한 뇌파는 알츠하이머 병 (AD)과 같은 신경 퇴행성 장애에 매우 중요한 영향을 미칩니다. 연구에 따르면 AD 환자의 변화된 뇌파 기록 및 진동 성 네트워크의 손상이 흔한 것으로 나타났습니다. 그러나 신경 퇴행성 질환에서의 네트워크 진동의 병태 생리에 관한 우리의 지식은놀랍게도 불완전하고 더 탐험해야 할 필요가있다. 이 프로토콜에서는 정상 뇌와 병리학 뇌 모두에서 전기 통신을 이해할 수 있도록 뇌파를 기록 할 수있는 간단한 전극을 설계했습니다. 이 방법의 외과 이식은 다른 가용 절차보다 저렴합니다 7 . 또한,이 방법은보다 긴 시간 프레임 ( 즉, 하루 2 ~ 4 시간) 동안 고품질의 안정적인 뇌파 신호를 기록하는 데 사용할 수 있습니다. 또한 우리는 더 자연스럽게 행동 할 수있는 더 가벼운 전극 (무게 약 26mg)을 사용했습니다. 이 방법은 전기 생리학 실험실에서 일반적으로 사용되는 증폭기 및 디지타이저가 필요한 신생아 쥐의 뇌파 연구에 널리 적용 할 수 있으며 추가 장치가 필요하지 않습니다.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Computer pin</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Pincer</td>
			<td>DELI Group Co., Ltd.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>502 super glue</td>
			<td>DELI Group Co., Ltd.</td>
			<td>7144</td>
			<td></td>
		</tr>
		<tr>
			<td>Drying oven</td>
			<td>Boxun</td>
			<td>GZX-9140MBE</td>
			<td></td>
		</tr>
		<tr>
			<td>Isofluorane</td>
			<td>RWD Life Science</td>
			<td>902-0000-522</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotaxic apparatus</td>
			<td>RWD Life Science</td>
			<td>900-0068-507</td>
			<td></td>
		</tr>
		<tr>
			<td>Anesthesia apparatus</td>
			<td>RWD Life Science</td>
			<td>902-0000-510</td>
			<td></td>
		</tr>
		<tr>
			<td>Homeothermic Heating Device</td>
			<td>Harvard Apparatus</td>
			<td>K 024509</td>
			<td></td>
		</tr>
		<tr>
			<td>Amplifier Model 3000&#160;</td>
			<td>A-M Systems</td>
			<td>61558</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro1401 Analog Digital converter</td>
			<td>Cambridge Electronic Design Ltd.</td>
			<td>4383</td>
			<td>Data acquisition unit</td>
		</tr>
		<tr>
			<td>Spike2</td>
			<td>Cambridge Electronic Design Ltd.</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

recording eeg in freely moving neonatal rats using a novel method

뇌파는 뇌의 전기적 활동을 탐지하는 유용한 방법입니다. 또한, 간질이나 신경 퇴행성 장애와 같은 다양한 신경 학적 상태에 널리 사용되는 진단 도구입니다. 그러나 신생아에서 뇌파 기록을 얻는 것은 전문적인 취급과주의가 필요하기 때문에 기술적으로 어렵습니다. 여기, 신생아 쥐 (P8-P15)에서 EEG를 기록하는 새로운 방법을 제시합니다. 우리는 컴퓨터 핀 궤적을 사용하여 간단하고 신뢰할 수있는 전극을 설계했습니다. 정상 및 간질 두뇌에 고품질의 뇌파 신호를 기록하기 위해 쥐 강아지의 두개골에 쉽게 이식 할 수 있습니다. 새끼들은 간질 발작을 유도하기 위해 신경독 카이 닉 애씨드 (KA)를 복강 내 (ip) 주사 하였다. 이 수술에서 시행되는 외과 적 이식은 신생아의 다른 뇌파 검사보다 저렴합니다. 이 방법을 사용하면 1 주일 이상 동안 고품질의 안정적인 뇌파 신호를 기록 할 수 있습니다. 또한,이 과정은 성인 ra에도 적용될 수 있습니다간질이나 다른 신경계 질환을 연구하기 위해 ts와 마우스를 사용합니다.

<html> 위와 같은 외과 수술이 적절하게 시행되면, 한 쥐의 신생아 강아지 뇌파 기록이 성공적으로 수행됩니다. KA 주입 10 분 후 규칙적인 행동 신호 패턴이 불규칙한 움직임과 긁힘, 떨림, 균형 감퇴의 형태로 나타났습니다. 그림 2는 대표적인 생 EEG 추적과 발작 발작, 발작 - 강장 및 발작 - 발톱 확장 된 추적을 보여줍니다. 재발 성 발작과 발작 뇌파 방전 패턴은 KA 주입 후 15...

Watch this Scientific Journal Video about Recording EEG in Freely Moving Neonatal Rats Using a Novel Method at JoVE.com

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

여기서는 자유롭게 움직이는 신생아 간질 강아지에게 뇌파 검사 (EEG)를 기록하고 그 절차, 특징 및 적용을 설명하기 위해 고안된 새로운 기술을 소개합니다. 이 방법으로 1 주일 이상 뇌파를 기록 할 수 있습니다.

새로운 방법으로 자유롭게 움직이는 신생아 쥐의 뇌파 기록

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

recording-eeg-in-freely-moving-neonatal-rats-using-a-novel-method

Research

JoVE Journal

Neuroscience

13.1K 조회수.  South China Normal University.  여기서는 자유롭게 움직이는 신생아 간질 강아지에게 뇌파 검사 (EEG)를 기록하고 그 절차, 특징 및 적용을 설명하기 위해 고안된 새로운 기술을 소개합니다. 이 방법으로 1 주일 이상 뇌파를 기록 할 수 있습니다. 

비디오: Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

Novel Apparatus and Method for Drug Reinforcement

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction. Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in animal research to model various components of drug reinforcement, consumption, and addiction in humans. For this study, we used a novel approach to studying drug reinforcement in rats by combining traditional CPP and self-administration methodologies. We assembled an apparatus using two Med Associate operant chambers, sensory stimuli, and a Plexiglas-constructed neutral zone. These modifications allowed our experiments to encompass motivational aspects of drug intake through self-administration and drug-free assessment of drug/cue conditioning strength with the CPP test. In our experiments, rats self-administered cocaine (0.75 mg/kg/inj, i.v.) during either four (e.g., the "short-term") or eight (e.g., the "long-term") alternating-day sessions in an operant environment containing distinctive sensory cues (e.g., olfactory and visual). On the alternate days, in the other (differently-cued) operant environment, saline was available for self-infusion (0.1 ml, i.v.). Twenty-four hours after the last self-administration/cue-pairing session, a CPP test was conducted. Consistent with typical CPP findings, there was a significant preference for the chamber associated with cocaine self-administration. In addition, in animals undergoing the long-term experiment, a significant positive correlation between CPP magnitude and the number of cocaine-reinforced lever responses. In conclusion, this apparatus and approach is time and cost effective, can be used to examine a wide array of topics pertaining to drug abuse, and provides more flexibility in experimental design than CPP or self-administration methods alone.

Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in research to model various components of drug reinforcement, consumption, and addiction in humans. In this report, we combined traditional CPP and self-administration methods as a novel approach to studying drug reinforcement and addiction in rats.

약물 보강 소설 장치 및 방법

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction.  Operant drug ...

연구

신경과학

High-density EEG Recordings of the Freely Moving Mice using Polyimide-based Microelectrode

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a noninvasive brain monitoring tool in cognitive neuroscience as well as a diagnostic tool for epilepsy and sleep disorders in neurology. However, the underlying mechanism of EEG rhythm generation is still under the veil. Recently introduced polyimide-based microelectrode (PBM-array) for high resolution mouse EEG1 is one of the trials to answer the neurophysiological questions on EEG signals based on a rich genetic resource that the mouse model contains for the analysis of complex EEG generation process. This application of nanofabricated PBM-array to mouse skull is an efficient tool for collecting large-scale brain activity of transgenic mice and accommodates to identify the neural correlates to certain EEG rhythms in conjunction with behavior. However its ultra-thin thickness and bifurcated structure cause a trouble in handling and implantation of PBM-array. In the presented video, the preparation and surgery steps for the implantation of PBM-array on a mouse skull are described step by step. Handling and surgery tips to help researchers succeed in implantation are also provided.

In this article, we described the surgery procedure and handling tips for implantation of ultra-thin polyimide-based microelectrode array (PBM-array) on the mouse skull for acquisition of high-density encephalography (EEG) in a mouse model.

Polyimide 기반 Microelectrode를 사용하여 자유롭게 이동 마우스의 고밀도 뇌파 레코딩

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a ...

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

쥐에서 자당의 갈망의 보육을 평가하기위한 일반적인 방법

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed single-neuron activity while animals are acquiring an operantly conditioned response, or when that response is extinguished. But even in these cases, observation periods usually encompass only a single stage of learning, i.e. acquisition or extinction, but not both (exceptions include protocols employing reversal learning; see Bingman&#160;et al.1 for an example). However, acquisition and extinction entail different learning mechanisms and are therefore expected to be accompanied by different types and/or loci of neural plasticity.
Accordingly, we developed a behavioral paradigm which institutes three stages of learning in a single behavioral session and which is well suited for the simultaneous recording of single neurons&#39; action potentials. Animals are trained on a single-interval forced choice task which requires mapping each of two possible choice responses to the presentation of different novel visual stimuli (acquisition). After having reached a predefined performance criterion, one of the two choice responses is no longer reinforced (extinction). Following a certain decrement in performance level, correct responses are reinforced again (reacquisition). By using a new set of stimuli in every session, animals can undergo the acquisition-extinction-reacquisition process repeatedly. Because all three stages of learning occur in a single behavioral session, the paradigm is ideal for the simultaneous observation of the spiking output of multiple single neurons. We use pigeons as model systems, but the task can easily be adapted to any other species capable of conditioned discrimination learning.

Learning new stimulus-response associations engages a wide range of neural processes which are ultimately reflected in changing spike output of individual neurons. Here we describe a behavioral protocol allowing for the continuous registration of single-neuron activity while animals acquire, extinguish, and reacquire a conditioned response within a single experimental session.

자유롭게 학습의 세 단계에 걸쳐 비둘기를 이동에서 단일 신경 세포 &#39;활동 전위를 기록

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed ...

Extracellular Wire Tetrode Recording in Brain of Freely Walking Insects

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform natural behavior. We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allowed us to record activity from multiple neurons simultaneously while a tethered cockroach turned or altered walking speed. While a major advance, tethered preparations provide access to limited behaviors and often lack feedback processes that occur in freely moving animals. We now present a modified version of that technique that allows us to record from the central complex of freely moving cockroaches as they walk in an arena and deal with barriers by turning, climbing or tunneling. Coupled with high speed video and cluster cutting, we can now relate brain activity to various parameters of the movement of freely behaving insects.

We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allows us to monitor activity in individual units of tethered cockroaches. Here we present a modified version of that technique that allows us to also record brain activity in freely moving insects.

자유롭게 곤충 워킹 뇌 세포 와이어 사극 기록

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform ...

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees (Bombus terrestris)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from flowering plants. Only a few studies have described the gustatory responses of bees toward toxins in food, and these experiments have mainly used the proboscis extension response on restrained honey bees. Here, a new behavioral assay is presented for measuring the feeding responses of freely-moving, individual worker bumble bees to nutrients and toxins. This assay measures the amount of solution ingested by each bumble bee and identifies how tastants in food influence the microstructure of the feeding behavior.
The solutions are presented in a microcapillary tube to individual bumble bees that have been previously starved for 2-4 hr. The behavior is captured on digital video. The fine structure of the feeding behavior is analyzed by continuously scoring the position of the proboscis (mouthparts) from video recordings using event logging software. The position of the proboscis is defined by three different behavioral categories: (1) proboscis is extended and in contact with the solution, (2) proboscis is extended but not in contact with the solution and (3) proboscis is stowed under the head. Furthermore the speed of the proboscis retracting away from the solution is also estimated.
In the present assay the volume of solution consumed, the number of feeding bouts, the duration of the feeding bouts and the speed of the proboscis retraction after the first contact is used to evaluate the phagostimulatory or the deterrent activity of the compounds tested.
This new taste assay will allow researchers to measure how compounds found in nectar influence the feeding behavior of bees and will also be useful to pollination biologists, toxicologists and neuroethologists studying the bumble bee&#39;s taste system.

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees Bombus terrestris

A novel behavioral assay is described for investigating the short term gustatory responses of the mouthparts of freely-moving bumble bees (Bombus terrestris) toward nutrients and toxins in solution.

소설 행동 분석은 (엉망으로 꿀벌을 자유롭게 이동 개인의 미각 응답을 조사하기<em&gt; 뒤의 terrestris</em&gt;)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from ...

Noninvasive EEG Recordings from Freely Moving Piglets

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel telemetric electroencephalography system in combination with standard self-adhesive hydrogel electrodes. The piglets are calmed down without the use of sedatives. After their release into the pigpen, the piglets behave normally&#8212;they drink and sleep in the same cycle as their siblings. Their sleep phases are used for the EEG recordings.

Here, we present a protocol to record telemetric electroencephalograms (EEGs) from freely moving piglets directly in the pigpen without the use of a sedative, making it possible to record typical EEG patterns during non-REM sleep, like spindle bursts.

자유롭게 이동 Piglets에서 비 침 투 적인 뇌 파 기록

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel ...

Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain interstitial space with the behavior and/or with the specific outcome of a pathology (e.g., seizures for epilepsy). When studying epilepsy, the microdialysis technique is often combined with short-term or even long-term video-electroencephalography (EEG) monitoring to assess spontaneous seizure frequency, severity, progression and clustering. The combined microdialysis-EEG is based on the use of several methods and instruments. Here, we performed in vivo microdialysis and continuous video-EEG recording to monitor glutamate and aspartate outflow over time, in different phases of the natural history of epilepsy in a rat model. This combined approach allows the pairing of changes in the neurotransmitter release with specific stages of the disease development and progression. The amino acid concentration in the dialysate was determined by liquid chromatography. Here, we describe the methods and outline the principal precautionary measures one should take during in vivo microdialysis-EEG, with particular attention to the stereotaxic surgery, basal and high potassium stimulation during microdialysis, depth electrode EEG recording and high-performance liquid chromatography analysis of aspartate and glutamate in the dialysate. This approach may be adapted to test a variety of drug or disease induced changes of the physiological concentrations of aspartate and glutamate in the brain. Depending on the availability of an appropriate analytical assay, it may be further used to test different soluble molecules when employing EEG recording at the same time.

Here, we describe a method for in vivo microdialysis to analyze aspartate and glutamate release in the ventral hippocampus of epileptic and non-epileptic rats, in combination with EEG recordings. Extracellular concentrations of aspartate and glutamate may be correlated with the different phases of the disease.

자유롭게 이동 하는 쥐에서 뇌 파 기록 하는 동안 흥분 성의 아미노산의 Microdialysis

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain ...

Hybrid Microdrive System with Recoverable Opto-Silicon Probe and Tetrode for Dual-Site High Density Recording in Freely Moving Mice

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, conventional microdrive designs often only allow use of one type of electrode to record from single or multiple regions, limiting the yield of single-unit or depth profile recordings. It also often limits the ability to combine electrode recordings with optogenetic tools to target pathway and/or cell type specific activity. Presented here is a hybrid microdrive array for freely moving mice to optimize yield and a description of its fabrication and reuse of the microdrive array. The current design employs nine tetrodes and one opto-silicon probe implanted in two different brain areas simultaneously in freely moving mice. The tetrodes and the opto-silicon probe are independently adjustable along the dorsoventral axis in the brain to maximize the yield of unit and oscillatory activities. This microdrive array also incorporates a set-up for light, mediating optogenetic manipulation to investigate the regional- or cell type-specific responses and functions of long-range neural circuits. In addition, the opto-silicon probe can be safely recovered and reused after each experiment. Because the microdrive array consists of 3D-printed parts, the design of microdrives can be easily modified to accommodate various settings. First described is the design of the microdrive array and how to attach the optical fiber to a silicon probe for optogenetics experiments, followed by fabrication of the tetrode bundle and implantation of the array into a mouse brain. The recording of local field potentials and unit spiking combined with optogenetic stimulation also demonstrate feasibility of the microdrive array system in freely moving mice.

This protocol describes the construction of a hybrid microdrive array that allows implantation of nine independently adjustable tetrodes and one adjustable opto-silicon probe in two brain regions in freely moving mice. Also demonstrated is a method for safely recovering and reusing the opto-silicon probe for multiple purposes.

자유롭게 움직이는 마우스에서 이중 사이트 고밀도 레코딩을 위한 회수 가능한 광실리콘 프로브 및 Tetrode가 있는 하이브리드 마이크로드라이브 시스템

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, ...

Multi-Fiber Photometry to Record Neural Activity in Freely-Moving Animals

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and smaller functional subgroups, it becomes paramount to record from projections and/or genetically-defined subpopulations of neurons. Fiber photometry is an accessible and powerful approach that can overcome these challenges. By combining optical and genetic methodologies, neural activity can be measured in deep brain structures by expressing genetically-encoded calcium indicators, which translate neural activity into an optical signal that can be easily measured. The current protocol details the components of a multi-fiber photometry system, how to access deep brain structures to deliver and collect light, a method to account for motion artifacts, and how to process and analyze fluorescent signals. The protocol details experimental considerations when performing single and dual color imaging, from either single or multiple implanted optic fibers.

This protocol details how to implement and perform multi-fiber photometry recordings, how to correct for calcium-independent artifacts, and important considerations for dual-color photometry imaging.

자유롭게 움직이는 동물의 신경 활동을 기록하는 다중 섬유 광도 측정법

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and ...