この研究は、中国の自然科学財団（31171355）および広東の自然科学財団（S2011010003403,2014A030313440）によって支持された。

動物のケア、外科手術および記録手順は、南中国師範大学の動物ケアおよび使用委員会のガイドラインに従った。 電極調製（図1A〜C） 注記：コンピュータピンの軌跡は、通信デバイスの信号インタフェースの一部としての単なる接触です。それは、雌コネクタに差し込まれる雄コネクタからなる。 <ol><li>ピンサーの助けを借りて、雄と雌のピンをコンピュータピンの軌跡（ 図1A ）から慎重に分離します。オスとメスのピンを一緒に接続して電極を形成し、シアノアクリレートを塗布して強力な接着結合を作ります（ 図1C ）。 </li><li>電極を蒸留水で満たしたビーカーに入れ、超音波洗浄機に10分間置いてください。 45℃の乾燥オーブンに30分間移動してください。 UV光を用いて30分間電極を滅菌する。 </li></ol>手術手順（図1D〜F） <ol><li>滅菌された外科用器具および定位固定装置を準備する。空気でイソフルラン麻酔（2.5％）を用いて新生児ラットの仔犬を麻酔する。子犬が深く麻酔をかけられたら、イソフルランの用量を1.0％に調整します。適切な麻酔深度を確保するために、手術前に尾またはつまみを締めてください。 </li><li>イヤーバーを外耳道の中に入れ、わずかに締め付けることによって、定位固定装置内の幼児の頭を固定する。 注：新生児の頭蓋骨は非常に柔らかいので、イヤーバーを過度に締めすぎないでください。 </li><li>すべての装置に70％エタノールを噴霧することにより、無菌の手術野を維持する。メスを使用して頭部に15 mmの切開を行います。鉗子を使用して、静かに四隅の正中線から頭皮を引き離します。切開部を広く開いたままにするために、生理食塩水に浸した綿を皮膚の下に置きます（ 図1D ）。 </li><li>頭蓋骨のブレグマとラムダ点を見つけるそれらを鉛筆でマークしてください。シリンジ針（26G）を使用して、前頭前野皮質（PFC）と海馬に2つの穴をあけます。 注：Pregは、ブレグマの後ろ1.8mmおよび正中線の-0.5mmの側に位置するが、海馬はブレグマの前-2.0mmおよび正中線の±0.5mmに位置する（ 図1DおよびE ）。脳の損傷を最小限に抑えるために、電極の深さは皮質表面の2mmを超えてはならない。 </li><li>電極を保持するために鉗子を使用し、PFCおよび海馬に参照電極および記録電極をそれぞれ挿入する。エリスロマイシン軟膏を電極の周囲に塗布して、感染を避ける。シアノアクリレートを用いて電極を固定する。 </li><li>歯科用アクリルセメントを準備して、粘りと粘りのある一貫性があるようにします。電極と頭蓋骨の残りの部分を覆うように歯科用セメントを塗布します。 注：歯科用セメントを塗布する前に、頭蓋骨を完全に乾燥させます。 </li&gt; <li> 5％ピクリン酸を電極に塗布して保護します。 注：滅菌条件を維持するためには、手順全体をバイオセーフティフードで行う必要があります。 </li><li>定位固定フレームから動物を取り出し、300μLの10％グルコースを皮下注射する。加熱した毛布に置き、回復させます。動物が暖かい（37℃）、歩行可能である（ すなわち、完全に回復した）ことを確認する。手術後の痛みに対してブプレノルフィンを腹腔内投与する（0.05mg / kg）。 注意：十分な意識（すなわち、正常な行動や動き）を回復するまで、動物を放置しないでください。 </li><li>意識を取り戻した後、ダムを持ってその家のケージに子犬を戻してください。動物が完全に回復するまで2日間待ちます。 </li></ol> 3.脳波記録<ol><li>完全に回復した後、仔犬の頭蓋骨に埋め込まれた電極を、それ自身のケージ内の増幅器に接続する。アンプをアナログ/デジタル変換器に接続します。コンバータをコンピュータに接続します。接続線は絡み合わないように注意深く取り扱わなければなりません。 </li><li>記録のためにデータ取得ユニットで少なくとも10,000 Hzのサンプリングレートを選択します（トランスミッタの帯域幅は1〜100 Hzです）。データが適切にサンプリングされていることを確認してください。 </li><li>ベースライン記録を得た後、カイニン酸（KA）（2mg / kg）を腹腔内に注射して、てんかん発作を誘発する。 KA注射の15分後に、てんかん発作を観察し、記録する。 KAによる発作は通常肉体的である。 注：発作持続時間は約15.2±0.9秒であり、発作持続時間は約62±5秒である。新生仔ラットでは、抱水クロラール（400 mg / kg）をIP注射することにより発作を予防することができます。 </li><li>デジタル化されたデータを保存し、spike2などの信号処理ソフトウェアパッケージを使用して解析します。 perfによって新生児のEEG信号の異なる周波数成分のパワーレベルを明らかにするパワースペクトル分析を行うこと。 1から100 Hz（EEGバンド）までの二乗平均平方根振幅を求めることによって、1分の時間枠でパワーを計算する。 </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>

著者らは、利害の対立を宣言していない。

ここでは、自由に動く新生児ラットの仔犬をEEGで取得するための手術と記録の手順を有線方式で報告する（ 図3 ）。 P7-P12ラットの子犬は、満期のヒト新生児に対応する発達年齢であることが示唆されている10,11 。この年齢層のラットの仔犬と一緒に作業する場合、高品質のEEG記録データを得ることは技術的に困難です。さらに特殊な取り扱いと大きな注意が必?...

正常な脳機能を得るためには、ニューロン間の連続的な通信が必要であることは十分に確立されている。神経細胞間通信は、主にシナプスで行われ、シナプスでは、1つのニューロンからの情報が第2のニューロンに伝達される。このシナプス伝達は、電気的または化学的シナプスの2種類の専用構造配置によって媒介される。電気生理学は、身体全体の機能や行動を制御する、インターナルコミュニケーションの間に生成される電位を捕捉する分野です。 EEGは、多くの電気生理学的手法の中で最も一般的に使用される方法である。 EEGは、内部刺激または外部刺激によって生成された電気信号の変化を検出するために使用される技術である。さらに、それは、てんかん、パーキンソン病およびアルツハイマー病などの様々な神経学的状態の臨床診断および予後予測のための必須試験であるeならびに薬理学的および毒物学的作用物質の影響3 。一般に、てんかん患者は、過興奮性および脳内の機能的連結性を損なう。これらは、間質性てんかん様放電（IED）として要約され、鋭く過渡的なスパイクの形態でEEGによって記録することができる。鋭い波;スパイク波複合体;またはポリスピーク4 。てんかんの脳の主な特徴は、発作5の原因となる脳領域の位置を特定するために、頭皮または脳実質から記録できる、てんかん発作の自発的発生である5 。さらに、脳波はまた、アルツハイマー病（AD）のような神経変性障害において非常に重要な意味を有する。研究によれば、AD患者の脳波記録の変化および振動ネットワークの障害が一般的であることが示唆されている。しかしながら、神経変性疾患iにおけるネットワーク振動の病態生理学に関する我々の知識驚くほど不完全で、さらに調査する必要がある6 。 このプロトコルでは、正常脳と病理学的脳の両方における電気通信を理解するためにEEGを記録することができる簡単な電極を設計した。この方法における外科移植は、他の利用可能な手順7より安価である7 。さらに、この方法は、より長い時間枠（ すなわち、毎日2〜4時間、1週間）の高品質で安定した脳波信号を記録するために使用することができる。さらに、私たちは動物の自然な行動を可能にする軽い電極（体重約26mg）を使用しました8 。この方法は、電気生理学研究室で一般的に使用されるアンプとデジタイザを必要とする新生児ラットの仔犬の脳波検査に広く適用され、追加のデバイスは必要ありません。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
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			<td>Computer pin</td>
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			<td>DELI Group Co., Ltd.</td>
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			<td>DELI Group Co., Ltd.</td>
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			<td>RWD Life Science</td>
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			<td>RWD Life Science</td>
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			<td>Anesthesia apparatus</td>
			<td>RWD Life Science</td>
			<td>902-0000-510</td>
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			<td>Homeothermic Heating Device</td>
			<td>Harvard Apparatus</td>
			<td>K 024509</td>
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			<td>Amplifier Model 3000&#160;</td>
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			<td>4383</td>
			<td>Data acquisition unit</td>
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			<td>Spike2</td>
			<td>Cambridge Electronic Design Ltd.</td>
			<td></td>
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		</tr>
	</tbody>
</table>

recording eeg in freely moving neonatal rats using a novel method

脳波は脳内の電気的活動を検出するのに有用な方法です。さらに、それは、てんかんおよび神経変性疾患などの様々な神経学的状態のための広く使用される診断ツールである。しかし、特殊な取り扱いと大きな注意が必要なため、新生児の脳波記録を取得することは技術的に困難です。ここでは、新生児ラットの仔（P8〜P15）にEEGを記録するための新しい方法を紹介します。我々はコンピュータのピン軌跡を用いて簡単で信頼性の高い電極を設計した。ラットの頭蓋骨に簡単に移植することができ、高品質の脳波信号を正常およびてんかんの脳に記録することができます。仔は、癲癇発作を誘発するために神経毒カイニン酸（KA）の腹腔内（ip）注射を与えられた。この手技で実施される外科移植は、新生児の他のEEG処置よりも安価である。この方法により、高品質で安定した脳波信号を1週間以上記録することができます。さらに、この手順は成人RAにも適用できるてんかんまたは他の神経学的障害を研究するために使用される。

<html>上記の外科的処置が適切に行われれば、1チャンネルのラットの新生児の脳波記録が首尾よく行われる。 KA注射の10分後に、規則的な行動徴候のパターンが、不規則な動きおよび掻痒、振戦、およびバランスの喪失の形態で現れた。図2は、代表的な生EEGトレースおよび発作間欠性、発作性痙攣性および発作性発作性拡張性痕跡を示す。 KA注入後、15〜60分（30±5.2分;...

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

ここでは、自由に動く新生児のてんかんの仔に脳波記録（EEG）を記録し、その手順、特徴、および用途を記載するように設計された新規技術を紹介する。この方法は、1週間以上のEEGを記録することを可能にする。

新しい方法を用いた自由に動く新生児ラットの脳波記録

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 ビュー.  South China Normal University. ここでは、自由に動く新生児のてんかんの仔に脳波記録（EEG）を記録し、その手順、特徴、および用途を記載するように設計された新規技術を紹介する。この方法は、1週間以上のEEGを記録することを可能にする。 

ビデオ: 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.

薬物の強化のための新規装置および方法

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

神経科学

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.

ポリイミドベースのマイクロ電極を用いて自由に動くマウスの高密度脳波録音

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.

ラットのショ糖の渇望のインキュベーションを評価するための一般的な方法

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.

自由に学習の三つの段階を越えハトを移動する単一ニューロン」の活動電位を記録する

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.

自由に昆虫を歩くの脳における細胞外ワイヤーテトロード録音

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.

小説行動アッセイ個人の味覚応答を調査するために、自由に移動バンブル蜂（<em&gt;セイヨウオオマルハナバチ</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.

子豚を自由に移動するから非侵襲的脳波

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.

自由行動ラットの脳波記録中に興奮性アミノ酸の透析

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.

自由に動くマウスの二重部位高密度記録のための回収可能なオプトシリコンプローブとテトロードを備えたハイブリッドマイクロドライブシステム

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.

自由に動く動物の神経活動を記録するマルチファイバー光メトリー

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 ...