この研究は、NIHグラントR15DC013864およびR21DC011342（J. Irwin、PI）によって支持された。絵に登場する両親と個人は、ここに掲載されているすべての写真が出版目的で使用されることを許可しました。

<ol>
	<li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Diagnostic and statistical manual of mental disorders (5th ed). , (2013).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Diagnostic and statistical manual of mental disorders (4th ed). , (2000).</li><li>Beukelman, , Mirenda, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Augmentative and Alternative Communication. 4th Edition. , (2003).</li><li>Cameron, L., Murphy, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Obtaining+consent+to+participate+in+research:+The+issues+involved+in+including+people+with+a+range+of+learning+and+communication+disabilities.">Obtaining consent to participate in research: The issues involved in including people with a range of learning and communication disabilities.</a> Br J Learn Disabil. 35 (2), 113-120 (2006).</li><li>Cooper, J. O., Heron, T. E., Heward, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Applied behavior analysis, 2nd ed. , (2007).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Belmont report: Ethical principles and guidelines for the protection of human subjects of research. , (1978).</li><li>Dunn-Buron, K., Curtis, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The incredible 5-point scale: The significantly improved and expanded second edition. , (2012).</li><li>Gillott, A., Furniss, F., Walter, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anxiety+in+high-functioning+children+with+autism.">Anxiety in high-functioning children with autism.</a> Autism. 5 (3), 277-286 (2001).</li><li>Harwood, V., Preston, J. L., Grela, B., Harold, O., Andrada, K., Landi, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+correlates+of+speech+perception+in+toddlers:+Associations+among+ERPs,+nonword+repetition,+and+language.">Electrophysiological correlates of speech perception in toddlers: Associations among ERPs, nonword repetition, and language.</a> J Speech Lang Hearing Res. , (1995).</li><li>Hodgdon, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Visual strategies for improving communication. , (1995).</li><li>Irwin, J., Avery, T., Brancazio, L., Turcios, J., Ryherd, K., Landi, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+Indices+of+Audiovisual+Speech+Perception:+Beyond+the+McGurk+Effect+and+Speech+in+Noise.">Electrophysiological Indices of Audiovisual Speech Perception: Beyond the McGurk Effect and Speech in Noise.</a> Multisens Res. , .</li><li>Irwin, J. R., Tornatore, L. A., Brancazio, L., Whalen, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Can+children+with+autism+spectrum+disorders+"hear"+a+speaking+face.">Can children with autism spectrum disorders "hear" a speaking face.</a> Child Dev. 82 (5), 1397-1403 (2011).</li><li>Mancil, G. R., Pearl, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Restricted+Interests+as+Motivators:+Improving+Academic+Engagement+and+Outcomes+of+Children+on+the+Autism+Spectrum.">Restricted Interests as Motivators: Improving Academic Engagement and Outcomes of Children on the Autism Spectrum.</a> TEACHING Exceptional Children Plus. 4 (6), 2-12 (2008).</li><li>McPartland, J., Tillman, R., Yang, D., Bernier, R., Pelphrey, K., Volkmar, F., Rogers, S., Paul, R., Pelphrey, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+social+neuroscience+of+autism+spectrum+disorder.">The social neuroscience of autism spectrum disorder.</a> Handbook of autism and pervasive developmental disorders. 4 (1), 482-496 (2014).</li><li>Myles, B., Trautman, M., Schelvan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The hidden curriculum: Practical solutions for understanding unstated rules in social situations. , (2004).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Findings and conclusions: National standards project, phase 2. , (2015).</li><li>Nind, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conducting+qualitative+research+with+people+with+learning+communication+and+other+disabilities.">Conducting qualitative research with people with learning communication and other disabilities.</a> Methodological challenges. , (2009).</li><li>Premack, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reinforcement+theory.">Reinforcement theory.</a> Nebraska symposium on motivation. 13, 123-180 (1965).</li><li>Scattone, D., Wilczynski, S. M., Edwards, R. P., Rabian, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreasing+disruptive+behaviors+of+children+with+autism+using+social+stories.">Decreasing disruptive behaviors of children with autism using social stories.</a> J Autism Dev Disord. 3 (26), 535-543 (2002).</li><li>Thiemann, K. S., Goldstein, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+stories,+written+text+cues,+and+video+feedback;+Effects+on+social+communication+of+children+with+autism.">Social stories, written text cues, and video feedback; Effects on social communication of children with autism.</a> J Appl Behav Anal. 34 (4), 425-446 (2001).</li><li>Watson, D., Clark, L. A., Tellegen, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+validation+of+brief+measures+of+positive+and+negative+affect:+The+PANAS+scales.">Development and validation of brief measures of positive and negative affect: The PANAS scales.</a> J Pers Soc Psychol. 4 (6), 1063-1070 (1988).</li><li>Wong, C., 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=Evidence-based+practices+for+children,+youth+and+young+adults+with+Autism+Spectrum+Disorders:+a+comprehensive+review.">Evidence-based practices for children, youth and young adults with Autism Spectrum Disorders: a comprehensive review.</a> J Autism Dev Disord. 45 (7), (2013).</li></ol>

著者には報告すべき開示がない。

上記で概説した習熟プロトコルは、ASDを有する子供参加者が複数の試験セッションおよび脳波データ収集を完了する可能性を高めるために、EBPを適用する。この熟知プロセスのステップを実行する前に、参加者はEEG研究集録プロトコルのすべてのコンポーネントを完了できませんでした。この研究の予備データは、慣習化プロトコールに従事する全ての参加者が完全なEEGデータ収集を完了で?...

自閉症スペクトラム障害（ASDには）社会的コミュニケーション困難と動作1,2の制限された、繰り返しパターンのコンスタレーションによって特徴付けられる神経発達障害です。 ASDの基準を満たす特定された個体の数が増加するにつれて、ASDに関連する神経学的基礎をより良く理解したいという欲求も増している。専門家は、今後の研究では、非定型脳機能の早期発見のためのプロセスの開発を含めるべきであり、社会行動に影響を与えると仮定された脳システムを支援するための介入を導くべきであると示唆している14 。特に、この研究では、典型的な発達とASDなどのリスクがある小児のAV（Audio-Visual）知覚を評価するために、脳波検査（EEG）とイベント関連電位（ERP）開発アルバイト。 EEGとERPの両方の電気生理学的手法を使用することで、発達を通した典型的および非定型のAV音声知覚の両方に対する神経基盤のより良い理解がもたらされる。この研究アプローチは、連続的なEEGデータとERPの両方の分析を含む。 ERPはEEGデータの収集手順を検討するために、EEGの時間同​​期された複数の試行平均部分であるため、 この作業ができるだけ母集団を代表するようにするために、社会的、認知的、言語的能力の程度が異なるASD患者が参加者として含まれています。現在、高品質の脳波データを収集するシステムは、発達障害のある子供に大きな需要をもたらしています。 ASDを有する個人が困難を抱える行動の例には、新規な状況に適応すること、不快な感覚刺激（ すなわち 、EEGキャップを装着すること）を許容すること、静かに座っている。これらの困難は、個人の感覚処理の課題、社会的視点からの離脱、潜在的な認知障害、同一性の主張と関連している可能性がある。集合的に、これらの特性は、研究活動1を構成する必要な課題に従う子供の能力に影響を与える可能性がある。したがって、ASD児の参加を促進するために、子どもの強さを支持し、その限界に適応する研究プロトコールを開発することが重要である。目標は、ASDを患っている子供が、研究活動を遂行するのに苦労することなく、有用な神経生物学的データにつながるように支援することです。 現在の仕事は、視線とASD 11、12との子供を含む言語と社会的障害児における視聴覚スピーチへの神経応答の大規模な研究の一部です。プライマリー実験プロトコルは、複数の行動評価セッション、ならびに1つのシミュレートされたEEGデータ収集セッションおよび1つの実際のEEGデータ収集セッションを含む。 EEGとERPの処理と分析の詳細についてはここでは説明しません。 Harwood et al。 ERPデータ処理プロトコルについては、図9を参照のこと。 このペーパーの焦点は、子供（特にASDのような発達障害のある人）が神経生物学の研究に携わるという課題に取り組むために考案された習熟プロトコルです。このタイプの多くの研究研究と同様に、参加者はEEGデータ収集に参加することに加えて、様々な標準化された評価作業を完了しなければならない。新しい環境に対処したり、不快な感覚刺激を許容したり、長時間座ったりするときにASDを患う個人が直面する課題のために、研究活動の参加者の理解を深めるための戦略が開発されなければならない。増加する理解と代理は、すべての研究活動を成功裏に完了するための参加者の能力を高めるでしょう。重要なことに、このプロトコルは、各参加者が自分の参加に関する積極的な決定を下す明確な能力を持ち、実験を通して感情状態を監視することによって、継続的な同意が保証される複数の機会を提供する4 。 自閉症の国立プロフェッショナル開発センター、国立自閉症センターはASD 16、22を持つ個人のためのエビデンスに基づく実践の確立のための明確な基準を開発してきた二つの組織です。現在の研究プロジェクトのプロトコルは、これらの基準を使用して、研究活動に成功するために個人が直面する可能性のある課題に対処するための特定の介入を特定、選択、開発、実施します。スペックこれらの情報源からの介入には、社会的物語、視覚的スケジュールの形式による視覚的支援、ロールプレイング、およびモデリングが含まれる22 。さらに、このプロトコルにはプレマックの原理の適用が含まれている18 。 Premack原理は、報酬に対する個々の予想挙動との間の短い、明確な接続を提供する、高優先アクティビティ13、18と低優先の活性を、以下のことを含みます。例えば、子供が10分間質問に答えた場合、彼は10分間好きなおもちゃで遊ぶことができます。 以前の研究は、社会的な物語が、破壊的な行動を示すASDを患う子供のための効果的な介入であることを示唆している19 。ソーシャル・ナラティブは、社会的期待に関連してより多くの理解を提供する手段として、状況の背景に関する背景と情報を提供するs。ほとんどの社会的物語には、期待される活動、関係者の可能な視点、構造化活動中の他の人々とのやりとりの方法が含まれる（ここで使用される社会的物語の例については図1を参照）。社会的な物語は、不安を軽減し、新しい状況での期待を持った個人を支援することが示されている15 。 視覚的なスケジュールは、ASDの子供が新しい社会的状況に対処するのを助けるのに効果的です20 。スケジュールの枠内での視覚的支援は、将来の活動の予測を支援するために、ASDを持つ個人に静的な参照を提供します（ここで使用される視覚的スケジュールの例については図2を参照）。さらに、発生するアクティビティの削除は、時間の経過を理解するのを助け、セッションの具体的な視覚的表現を提供する。この視覚的表現がなければ、ASDを患う個体は、どのくらいの時間が経過し、どのくらいの時間が活動やセッションに残っているのかを概念化することはできません。参加者が自分の感情を表現することを可能にする視覚的支援は、参加者が異なる感情により容易に対処できるようにするのにも役立つ可能性がある（感情評価尺度の例については図3を参照）。 ロールプレイングとモデリングは、ASDを持つ個人の新しい状況を見て練習する機会を提供します。リハーサルのプロセスが期待される行動についての明快さを提供し、さらにこれらの新規シナリオ8、22のための不安を軽減します。モデリングを含むロールプレイングは、個人に、特定のアクティビティに参加するかどうかの選択を練習し決定する明確な機会を提供する（ 図5および図 6を参照してください）クマとチャンスキャップを着用してください）。 ASDおよび/または他の発達障害を有する個人によっては、研究課題を明確に理解するための成熟度または認知能力を発達させていないものもある。確かに、複雑な社会状況における理解の困難さとインフォームド・コンセントに及ぼす影響により、脆弱なグループの参加を取り巻く倫理的ジレンマが存在する17 。しかし、プロセスがより良い不当な圧力4、17せずにインフォームドコンセントを確保するために開発されています。例えば、簡単な説明の視覚的な記号と箇条書きがここに提供され、参加者は進行中のプロセスとして研究に参加することに同意するかどうか尋ねられます3 。発達障害を持つ個人の理解を促進するために、研究目標に沿っていても、倫理指針に沿って説得せずに参加したり、圧力を元に戻すことを選択した主題6 。

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			<td height="42" style="height:42px;width:216px;">Electroencephalography Net</td>
			<td style="width:209px;">Electrical&#160; Geodesics Incorporated</td>
			<td style="width:203px;">EGI 300 Series Amplifier</td>
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a familiarization protocol facilitates the participation of children with asd in electrophysiological research

このペーパーは、脳波記録（EEG）データおよび事象関連電位（ERP）を収集するためのより大きな研究プロトコルの不可欠な構成要素として使用される、慣れプロトコルの詳細な説明を含む。現在、高品質EEG / ERPデータの収集に利用可能なシステムは、自閉症スペクトラム障害（ASD）を有するものなど、発達障害を有する子供に大きな需要をもたらす。 ASDの子供は、新しい状況に適応すること、不快な感覚刺激に耐えること、そして静かに座ることが困難な場合があります。この慣れプロトコルは、研究参加者の知識と研究プロトコルの特定の活動と手順の理解を高めるために、EBP（Evidence-Based Practices）を使用しています。この熟知プロトコルのツールは、社会的な物語、視覚的スケジュール、プレマックの原理、ロールプレイング、およびモデリングです。この習熟プロトコルの目的は、理解を深め、子ども参加者の不安を潜在的に軽減することができ、EEG / ERPデータの収集のための研究プロトコルの成功の可能性がより高くなる。

視線および視聴覚音声知覚に関する一連の研究の一部として、同様のEEGプロトコルを用いて、ASDおよび平均IQ（n = 25,19男児、女児6名、平均年齢= 10.25歳）の25人の参加者の最初のコホートを採用した（ 図9 ）。この以前のコホートの子供の72％が脳波プロトコルを完了しました。現在のコホートでは、これまでに15人のASD患者が募集された（n = 15,11男児、4人...

Watch this Scientific Journal Video about A Familiarization Protocol Facilitates the Participation of Children with ASD in Electrophysiological Research at JoVE.com

A Familiarization Protocol Facilitates the Participation of Children with ASD in Electrophysiological Research

電気生理学的研究は、自閉症スペクトル障害（ASD）を含む発達障害のバイオマーカーを同定するための重要なツールであるが、これらの集団におけるデータ収集は困難なままである。この研究は、ASDを患う小児からEEGデータを収集する可能性を改善する脳波記録（EEG）を含む研究に付随する親密化プロトコルを提示する。

親密化プロトコルは、電気生理学的研究におけるASDを有する小児の参加を促進する

This paper includes a detailed description of a familiarization protocol, which is used as an integral component of a larger research protocol to collect ...

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8.2K ビュー.  Southern Connecticut State University. 電気生理学的研究は、自閉症スペクトル障害（ASD）を含む発達障害のバイオマーカーを同定するための重要なツールであるが、これらの集団におけるデータ収集は困難なままである。この研究は、ASDを患う小児からEEGデータを収集する可能性を改善する脳波記録（EEG）を含む研究に付随する親密化プロトコルを提示する。 

ビデオ: A Familiarization Protocol Facilitates the Participation of Children with ASD in Electrophysiological Research

Assessment of Cerebral Lateralization in Children using Functional Transcranial Doppler Ultrasound (fTCD)

There are many unanswered questions about cerebral lateralization. In particular, it remains unclear which aspects of language and nonverbal ability are lateralized, whether there are any disadvantages associated with atypical patterns of cerebral lateralization, and whether cerebral lateralization develops with age. In the past, researchers interested in these questions tended to use handedness as a proxy measure for cerebral lateralization, but this is unsatisfactory because handedness is only a weak and indirect indicator of laterality of cognitive functions1. Other methods, such as fMRI, are expensive for large-scale studies, and not always feasible with children2.
Here we will describe the use of functional transcranial Doppler ultrasound (fTCD) as a cost-effective, non-invasive and reliable method for assessing cerebral lateralization. The procedure involves measuring blood flow in the middle cerebral artery via an ultrasound probe placed just in front of the ear. Our work builds on work by Rune Aaslid, who co-introduced TCD in 1982, and Stefan Knecht, Michael Deppe and their colleagues at the University of M&uuml;nster, who pioneered the use of simultaneous measurements of left- and right middle cerebral artery blood flow, and devised a method of correcting for heart beat activity. This made it possible to see a clear increase in left-sided blood flow during language generation, with lateralization agreeing well with that obtained using other methods3.
The middle cerebral artery has a very wide vascular territory (see Figure 1) and the method does not provide useful information about localization within a hemisphere. Our experience suggests it is particularly sensitive to tasks that involve explicit or implicit speech production. The 'gold standard' task is a word generation task (e.g. think of as many words as you can that begin with the letter 'B') 4, but this is not suitable for young children and others with limited literacy skills. Compared with other brain imaging methods, fTCD is relatively unaffected by movement artefacts from speaking, and so we are able to get a reliable result from tasks that involve describing pictures aloud5,6. Accordingly, we have developed a child-friendly task that involves looking at video-clips that tell a story, and then describing what was seen.

Assessment of Cerebral Lateralization in Children using Functional Transcranial Doppler Ultrasound fTCD

Functional transcranial Doppler sonography (fTCD) is a simple and non-invasive ultrasound technique which can be used to assess the lateralization of cognitive functions, especially language, and is suitable for use with children.

機能性経頭蓋ドップラー超音波を使って子供の脳の側性化の評価（fTCD）

There are many unanswered questions about cerebral lateralization.  In particular, it remains unclear which aspects of language and nonverbal ability are ...

神経科学

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs through activation of their ion channels and neurotransmitter receptors, and process information in part through activity-dependent release of gliotransmitters. Furthermore, astrocytes constitute the main uptake system for glutamate, contribute to potassium spatial buffering, as well as to GABA clearance. These cells therefore constantly monitor synaptic activity, and are thereby sensitive indicators for alterations in synaptically-released glutamate, GABA and extracellular potassium levels. Additionally, alterations in astroglial uptake activity or buffering capacity can have severe effects on neuronal functions, and might be overlooked when characterizing physiopathological situations or knockout mice. Dual recording of neuronal and astroglial activities is therefore an important method to study alterations in synaptic strength associated to concomitant changes in astroglial uptake and buffering capacities. Here we describe how to prepare hippocampal slices, how to identify stratum radiatum astrocytes, and how to record simultaneously neuronal and astroglial electrophysiological responses. Furthermore, we describe how to isolate pharmacologically the synaptically-evoked astroglial currents.

The preparation of acute brain slices from isolated hippocampi, as well as the simultaneous electrophysiological recordings of astrocytes and neurons in stratum radiatum during stimulation of schaffer collaterals is described. The pharmacological isolation of astroglial potassium and glutamate transporter currents is demonstrated.

急性海馬スライスにおけるシナプス誘発アストログリアとニューロン活動の二重の電気生理学的記録

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

Forebrain Electrophysiological Recording in Larval Zebrafish

Epilepsy affects nearly 3 million people in the United States and up to 50 million people worldwide. Defined as the occurrence of spontaneous unprovoked seizures, epilepsy can be acquired as a result of an insult to the brain or a genetic mutation. Efforts to model seizures in animals have primarily utilized acquired insults (convulsant drugs, stimulation or brain injury) and genetic manipulations (antisense knockdown, homologous recombination or transgenesis) in rodents. Zebrafish are a vertebrate model system1-3 that could provide a valuable alternative to rodent-based epilepsy research. Zebrafish are used extensively in the study of vertebrate genetics or development, exhibit a high degree of genetic similarity to mammals and express homologs for ~85% of known human single-gene epilepsy mutations. Because of their small size (4-6 mm in length), zebrafish larvae can be maintained in fluid volumes as low as 100 &mu;l during early development and arrayed in multi-well plates. Reagents can be added directly to the solution in which embryos develop, simplifying drug administration and enabling rapid in vivo screening of test compounds4. Synthetic oligonucleotides (morpholinos), mutagenesis, zinc finger nuclease and transgenic approaches can be used to rapidly generate gene knockdown or mutation in zebrafish5-7. These properties afford zebrafish studies an unprecedented statistical power analysis advantage over rodents in the study of neurological disorders such as epilepsy. Because the "gold standard" for epilepsy research is to monitor and analyze the abnormal electrical discharges that originate in a central brain structure (i.e., seizures), a method to efficiently record brain activity in larval zebrafish is described here. This method is an adaptation of conventional extracellular recording techniques and allows for stable long-term monitoring of brain activity in intact zebrafish larvae. Sample recordings are shown for acute seizures induced by bath application of convulsant drugs and spontaneous seizures recorded in a genetically modified fish.

A simple method to record extracellular field potentials in the larval zebrafish forebrain is described. The method provides a robust in vivo read-out of seizure-like activity. This technique can be used with genetically modified zebrafish larvae carrying epilepsy-related genes or seizures evoked by administration of convulsant drugs.

ゼブラフィッシュ幼生における前脳電気生理学的記録

Epilepsy affects nearly 3 million people in the United States and up to 50 million people worldwide. Defined as the occurrence of spontaneous unprovoked ...

A Proboscis Extension Response Protocol for Investigating Behavioral Plasticity in Insects: Application to Basic, Biomedical, and Agricultural Research

Insects modify their responses to stimuli through experience of associating those stimuli with events important for survival (e.g., food, mates, threats). There are several behavioral mechanisms through which an insect learns salient associations and relates them to these events. It is important to understand this behavioral plasticity for programs aimed toward assisting insects that are beneficial for agriculture. This understanding can also be used for discovering solutions to biomedical and agricultural problems created by insects that act as disease vectors and pests. The Proboscis Extension Response (PER) conditioning protocol was developed for honey bees (Apis mellifera) over 50 years ago to study how they perceive and learn about floral odors, which signal the nectar and pollen resources a colony needs for survival. The PER procedure provides a robust and easy-to-employ framework for studying several different ecologically relevant mechanisms of behavioral plasticity. It is easily adaptable for use with several other insect species and other behavioral reflexes. These protocols can be readily employed in conjunction with various means for monitoring neural activity in the CNS via electrophysiology or bioimaging, or for manipulating targeted neuromodulatory pathways. It is a robust assay for rapidly detecting sub-lethal effects on behavior caused by environmental stressors, toxins or pesticides.
We show how the PER protocol is straightforward to implement using two procedures. One is suitable as a laboratory exercise for students or for quick assays of the effect of an experimental treatment. The other provides more thorough control of variables, which is important for studies of behavioral conditioning. We show how several measures for the behavioral response ranging from binary yes/no to more continuous variable like latency and duration of proboscis extension can be used to test hypotheses. And, we discuss some pitfalls that researchers commonly encounter when they use the procedure for the first time.

The Proboscis Extension Response (PER) conditioning protocol, developed for the honey bee (Apis mellifera), provides an ecologically-relevant and easily quantifiable means for studying several different mechanisms of learning in many insect species.

昆虫に行動可塑性を調査するためのテングザルの拡張レスポンスプロトコル：基本、生物医学および農業研究への応用

Insects modify their responses to stimuli through experience of associating those stimuli with events important for survival (e.g., food, mates, threats). ...

Protocol for Three-dimensional Confocal Morphometric Analysis of Astrocytes

As glial cells in the brain, astrocytes have diverse functional roles in the central nervous system. In the presence of harmful stimuli, astrocytes modify their functional and structural properties, a condition called reactive astrogliosis. Here, a protocol for assessment of the morphological properties of astrocytes is presented. This protocol includes quantification of 12 different parameters i.e. the surface area and volume of the tissue covered by an astrocyte (astrocyte territory), the entire astrocyte including branches, cell body, and nucleus, as well as total length and number of branches, the intensity of fluorescence immunoreactivity of antibodies used for astrocyte detection, and astrocyte density (number/1,000 &#181;m2). For this purpose three-dimensional (3D) confocal microscopic images were created, and 3D image analysis software such as Volocity 6.3 was used for measurements. Rat brain tissue exposed to amyloid beta1-40 (A&#946;1-40) with or without a therapeutic&#160;intervention was used to present the method. This protocol can also be used for 3D morphometric analysis of other cells from either in vivo or in vitro conditions.

Astrocytes in the CNS change their functional and structural properties in response to harmful stimuli. This report presents a protocol for assessment of three-dimensional astrocyte morphology in diseased conditions or after therapeutic interventions.

アストロサイトの三次元共焦点形態学的分析のためのプロトコル

As glial cells in the brain, astrocytes have diverse functional roles in the central nervous system. In the presence of harmful stimuli, astrocytes modify ...

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the molecular and cellular basis of higher brain functions. A whole-mount preparation of adult brains with well-preserved morphology is critical for such whole brain-based studies, but can be technically challenging and time-consuming. This protocol describes an easy-to-learn, one-step dissection approach of an adult fly head in less than 10 s, while keeping the intact brain attached to the rest of the body to facilitate subsequent processing steps. The procedure helps remove most of the eye and tracheal tissues normally associated with the brain that can interfere with the later imaging step, and also places less demand on the quality of the dissecting forceps. Additionally, we describe a simple method that allows convenient flipping of the mounted brain samples on a coverslip, which is important for imaging both sides of the brains with similar signal intensity and quality. As an example of the protocol, we present an analysis of dopaminergic (DA) neurons in adult brains of WT (w1118) flies. The high efficacy of the dissection method makes it particularly useful for large-scale adult brain-based studies in Drosophila.

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

The adult Drosophila brain is a valuable system for studying neuronal circuitry, higher brain functions, and complex disorders. An efficient method to dissect whole brain tissue from the small fly head will facilitate brain-based studies. Here we describe a simple, one-step dissection protocol of adult brains with well-preserved morphology.

大人のホールマウントの準備のためのシンプルなワンステップ解剖プロトコル<em&gt;ショウジョウバエ</em&gt;ブレインズ

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the ...

In Vitro Characterization of the Electrophysiological Properties of Colonic Afferent Fibers in Rats

Dysfunction of the colonic sensory nerves has been implicated in the pathophysiology of several common conditions, including functional and inflammatory bowel diseases and diabetes. Here, we describe a protocol for the in vitro characterization of the electrophysiological properties of colonic afferents in rats. The colorectum, with the intact pelvic ganglion (PG) attached, is removed from the rat; superfused with carbogenated Krebs solution in the recording chamber; and cannulated at the oral and anal ends to allow for distension. A fine nerve bundle emanating from the PG is identified, and the multiunit afferent nerve activity is recorded using a suction electrode. Distension of the colonic segment elicits gradual increases in multiunit discharge. A principal component analysis is conducted to differentiate the low-threshold, the high-threshold, and the wide-dynamic range afferent fibers. Chemical sensitivity of colonic afferents can be studied through the bath or intraluminal administration of test compounds. This protocol can be modified for application to other species, such as mice and guinea pigs, and to study the differences in the electrophysiological properties of thoracolumbar/hypogastric and lumbosacral/pelvic afferents of the descending colon in normal and pathological conditions.

In Vitro Characterization of the Electrophysiological Properties of Colonic Afferent Fibers in Rats

Abnormal sensory function underlies visceral pain and other symptoms of functional and inflammatory bowel diseases. A protocol for the electrophysiological recording of the colonic afferent nerves in an ex vivo rat colorectum preparation is presented here.

体外ラットの結腸求心性線維の電気生理学的特性の評価

Dysfunction of the colonic sensory nerves has been implicated in the pathophysiology of several common conditions, including functional and inflammatory ...

In Vivo Electrophysiological Measurement of the Rat Ulnar Nerve with Axonal Excitability Testing

Electrophysiology enables the objective assessment of peripheral nerve function in vivo. Traditional nerve conduction measures such as amplitude and latency detect chronic axon loss and demyelination, respectively. Axonal excitability techniques &#34;by threshold tracking&#34; expand upon these measures by providing information regarding the activity of ion channels, pumps and exchangers that relate to acute function and may precede degenerative events. As such, the use of axonal excitability in animal models of neurological disorders may provide a useful in vivo measure to assess novel therapeutic interventions. Here we describe an experimental setup for multiple measures of motor axonal excitability techniques in the rat ulnar nerve.
The animals are anesthetized with isoflurane and carefully monitored to ensure constant and adequate depth of anesthesia. Body temperature, respiration rate, heart rate and saturation of oxygen in the blood are continuously monitored. Axonal excitability studies are performed using percutaneous stimulation of the ulnar nerve and recording from the hypothenar muscles of the forelimb paw. With correct electrode placement, a clear compound muscle action potential that increases in amplitude with increasing stimulus intensity is recorded. An automated program is then utilized to deliver a series of electrical pulses which generate 5 specific excitability measures in the following sequence: stimulus response behavior, strength duration time constant, threshold electrotonus, current-threshold relationship and the recovery cycle.
Data presented here indicate that these measures are repeatable and show similarity between left and right ulnar nerves when assessed on the same day. A limitation of these techniques in this setting is the effect of dose and time under anesthesia. Careful monitoring and recording of these variables should be undertaken for consideration at the time of analysis.

In Vivo Electrophysiological Measurement of the Rat Ulnar Nerve with Axonal Excitability Testing

Axonal excitability techniques provide a powerful tool to examine pathophysiology and biophysical changes that precede irreversible degenerative events. This manuscript demonstrates the use of these techniques on the ulnar nerve of anesthetized rats.

生体内で軸索興奮性のテストでラットの尺骨神経の電気生理学的測定

Electrophysiology enables the objective assessment of peripheral nerve function in vivo. Traditional nerve conduction measures such as amplitude and ...

Studying Brain Function in Children Using Magnetoencephalography

Magnetoencephalography (MEG) is a non-invasive neuroimaging technique which directly measures magnetic fields produced by the electrical activity of the human brain. MEG is quiet and less likely to induce claustrophobia compared with magnetic resonance imaging (MRI). It is therefore a promising tool for investigating brain function in young children. However, analysis of MEG data from pediatric populations is often complicated by head movement artefacts which arise as a consequence of the requirement for a spatially-fixed sensor array that is not affixed to the child's head. Minimizing head movements during MEG sessions can be particularly challenging as young children are often unable to remain still during experimental tasks. The protocol presented here aims to reduce head movement artefacts during pediatric MEG scanning. Prior to visiting the MEG laboratory, families are provided with resources that explain the MEG system and the experimental procedures in simple, accessible language. An MEG familiarization session is conducted during which children are acquainted with both the researchers and the MEG procedures. They are then trained to keep their head still whilst lying inside an MEG simulator. To help children feel at ease in the novel MEG environment, all of the procedures are explained through the narrative of a space mission. To minimize head movement due to restlessness, children are trained and assessed using fun and engaging experimental paradigms. In addition, children's residual head movement artefacts are compensated for during the data acquisition session using a real-time head movement tracking system. Implementing these child-friendly procedures is important for improving data quality, minimizing participant attrition rates in longitudinal studies, and ensuring that families have a positive research experience.

This article introduces a child-friendly research protocol designed to improve data quality by reducing head movement during pediatric magnetoencephalography (MEG). We familiarize families with the MEG environment, train children to remain still using an MEG simulator, and correct for residual head movement artefacts using a real-time head movement detection system.

子供の脳磁図を用いた脳機能の研究

Magnetoencephalography (MEG) is a non-invasive neuroimaging technique which directly measures magnetic fields produced by the electrical activity of the ...

A Protocol for Transcranial Photobiomodulation Therapy in Mice

Transcranial photobiomodulation is a potential innovative noninvasive therapeutic approach for improving brain bioenergetics, brain function in a wide range of neurological and psychiatric disorders, and memory enhancement in age-related cognitive decline and neurodegenerative diseases. We describe a laboratory protocol for transcranial photobiomodulation therapy (PBMT) in mice. Aged BALB/c mice (18 months old) are treated with a 660 nm laser transcranially, once daily for 2 weeks. Laser transmittance data shows that approximately 1% of the incident red light on the scalp reaches a 1 mm depth from the cortical surface, penetrating the dorsal hippocampus. Treatment outcomes are assessed by two methods: a Barnes maze test, which is a hippocampus-dependent spatial learning and memory task evaluation, and measuring hippocampal ATP levels, which is used as a bioenergetics index. The results from the Barnes task show an enhancement of the spatial memory in laser-treated aged mice when compared with age-matched controls. Biochemical analysis after laser treatment indicates increased hippocampal ATP levels. We postulate that the enhancement of memory performance is potentially due to an improvement in hippocampal energy metabolism induced by the red laser treatment. The observations in mice could be extended to other animal models since this protocol could potentially be adapted to other species frequently used in translational neuroscience, such as rabbit, cat, dog, or monkey. Transcranial photobiomodulation is a safe and cost-effective modality which may be a promising therapeutic approach in age-related cognitive impairment.

Photobiomodulation therapy is an innovative noninvasive modality for the treatment of a wide range of neurological and psychiatric disorders and can also improve healthy brain function. This protocol includes a step-by-step guide to performing brain photobiomodulation in mice by transcranial light delivery, which can be adapted for use in other laboratory rodents.

マウスにおける経頭蓋 Photobiomodulation 療法のプロトコル

Transcranial photobiomodulation is a potential innovative noninvasive therapeutic approach for improving brain bioenergetics, brain function in a wide ...