FDは、統合失調症とうつ病に関する研究とカナダの精神医学研究財団からCPRF賞のための国民連合からの若手研究者賞によってサポートされていました。

1。実験的なデザイン、方法論的問題、及び刺激 （SMPの設計を示す図については図1を参照）SMPは2つのタスク、エンコードタスクと検索タスクで構成されており、脳活動の記録は、エンコーディングタスク中、またはエンコードと検索の両方の作業中に発生する可能性があります。 <img src="/files/ftp_upload/2433/2433fig1.jpg" alt="図1" /> 図1。SMPの設計の概略図。その活性がエンコード中に、その後忘れられた項目のためのより続いて記憶のために大きいの脳領域（R&gt; F）の符号化成功（ES）に関連付けられている。メモリ内のこの差は（DM）3が検索（ヒット数&gt;ミス）中に記録された脳活動の比較に基づいて測定されている場合同様に、それは検索成功（RS）に関連付けられている領域を識別します。 いくつかの重要な側面は、感情的な刺激でSMPを用いた研究で考慮する必要があります。 <ol><li>彼らは感情的なメモリ10、26、27日にこのように感情的な知覚に異なった効果を持つとこのような感情価と覚醒/強度、のような刺激の感情的性質、をコントロールすることが重要です。一般的に使用される感情的な刺激は、国際情動写真システム（個別行動計画）23、英語の単語のためのアフェ規範（ANEW）24からの単語からの写真です。 （25参照）刺激の感情的なプロパティを定義する個別行動計画の画像とANEW言葉は2つの主要な感情の次元（感情的な強さ- -快または不快感、そして覚醒の度合い、すなわち、 原子価 ）で制御されます。頻繁に感情研究で使用される刺激の別のカテゴリでは、感情表現と人間の顔、そのような私達フェイシャル影響を受けるプラットフォーム（の写真からのものになります<a href="http://www.paulekman.com">http://www.paulekman.com</a> （）とNimStim <a href="http://www.macbrain.org/">http://www.macbrain.org/~~V</a> ）を設定します。 </li><li>その操作もメモリの性能に影響を与える、それ故にこのプロトコルの汎用性に貢献するかもしれない他の要因は次のとおりaをエンコーディングタスクの種類の変化は、B.、（すなわち偶発的11、13、19または意図的な8、20）が使用エンコーディングの刺激の感情的な内容（感情的な8と、すなわち、11または主要な焦点13、19などの非感情的側面 ​​）に関するフォーカス、およびc （すなわちリコール8、11または認識12、20）使用される検索タスクのタイプ。さらに、刺激の持続時間は、メモリのパフォーマンス21を影響し、従ってそれは上記の要因に加え、同様に考慮する必要があります。 </li><li>このように人間の存在感と感情とニュートラル画像の視覚的な複雑さは11、その他の可能な交絡変数は、また気をつける必要があります。 </li><li>それは興味の条件のためのメモリの性能が両方の行動と脳イメージングデータの公正な統計的な比較を可能にする合理的な範囲内に収まることが重要です。例えば、私たちのキューさ、リコールの作業で、記憶や忘れられた試験の割合は約50％8、11でした。二選択肢認識タスクでこの性能レベルに近いチャンスになるしかし、、より高い全体的なパフォーマンスのレベルが期待されている。このケースでは、記憶と忘れ去ら条件との間の公正な比較は、ランダムに忘れて試行回数を同一視することを覚えて試験の小さい数を選択することによって達成することができます。さらに、メモリ上に感情の増強効果は、また中立の項目よりも感情のための記憶試験の大部分に反映され、行動データに存在する必要があります。 </li><li>また、これもパフォーマンスに22を影響を与える可能性があると気分誘導が回避されることをお勧めします。これは、擬似ランダムと同じ価数のこれ以上の2から3以上の刺激が連続して11を提示していることを保証するために感情的な刺激をすることによって行うことができます。 </li></ol> 2。スキャンのために件名の準備すべての被験者は、事前の倫理委員会によって承認された実験的なプロトコルを、実行するためにインフォームドコンセントを書かれています。 前にスキャン部屋に入る<ol><li>約45分前にスキャンするために、各被験者は、全体的な感情の状態および不安レベル28、29を評価するアンケートを完了します。これらの初期評価は、研究に参加した結果として気分/不安の変化を画面にセッション後の評価と組み合わせて使用​​されています。 </li><li>アンケート終了後、被験者は、スキャン手順が通知さと行動のタスクのための具体的な指示を与えられます。主題はまた、タスクに慣れるために、短い練習セッションを完了します。 </li></ol> Scanniの入力NGルーム<ol start="3"><li>被験者は、頭部のための追加のクッションと、スキャニングベッドに仰臥位になるように指示されており、耳の保護が与えられます。また、テープの長さの非粘着面は、動きを最小限にするために被験者の前頭部の周りに軽くラップされることがあります。 </li><li>スキャンする前に、被験者の右手はレスポンスボックスに配置されていると応答ボックスがテストされます。データの収集を開始する前に、それはまた、被写体がはっきりと画面の投影を見ることができることを確保すべきである。 S /彼はスキャナを停止するあらゆる緊急の必要性を示すことができるように緊急停止ボタンは、近くの被写体のフリーハンドを配置されます。 </li><li>我々の研究では、刺激がCIGAL、社内データのプレゼンテーションソフトウェアプログラム30が、他のソフトウェアを使用して発表されたE -プライム、SuperLab、プレゼンテーション、およびMATLABベースのPsychtoolbox含め、もご利用いただけます。 </li><li>また、後者は臨床群17に削減する必要がある、可能であれば、当社の調査に基づいて、我々は、感情の刺激を含む、スキャンタスクの継続時間が1時間を超えるべきではない、と全体のスキャン時間が2時間を超えてはならないことをお勧めします。 </li></ol> 3。データの記録と処理スキャンパラメータ Dolcosと同僚11による最初の研究では、MRIデータは1.5テスラ、GEのスキャナーで採取された。 TE = 40msの; FOVフル脳報道は軸平面におけるAC - PCプレーン（TR = 3000msに平行に定義された、34の連続した​​グラディエントエコーechoplanar画像（EPIs）を経由して血中酸素濃度に敏感に依存（BOLD）コントラストを取得しました= 64 2画像行列、FA = 90 °、スライス厚= 3.75 mm）である。ここで使用されるスキャンパラメータは、よく内側側頭葉（MTL）の構造でアクティベーションを識別するのに適しているが、より高い空間分解能に最適化された他のスキャンパラメータは、33から35を使用することができます。 後のスキャンの手順<ol><li>被験者は、スキャンセッションの前に実行されない場合、性格特性（例えば神経症的傾向、感情的なarousability）の評価も収集することができる、状態の気分障害や不安のレベルを評価するアンケートを完了し。 </li><li>復旧活動は、スキャンセッション中に記録されていない場合、検索タスクは、スキャンセッションの後に実行するか、後ですることができます。すでにスキャナで実行されていない場合は、感情的な評価タスクは、刺激に対する被験者の感情的な反応を評価するために使用されることがあります。この方法では、感情的で中立的なカテゴリの分離は、参加者の格付けに基づいて行わはなく、規範的なスコアに基づいて行うことができます。しかし、このタスクは、検索タスクは、次を実行する必要があります。 </li><li>メモリ上に感情の影響を最大にするために、〜20分の最小遅延は符号化と検索の18日の間に発生する必要があります。メモリ上に感情の影響は長期的かつ感情的な記憶になることができるしかし、遅延は、数分から日、週、月に、はるかに長くすることができます中立回想12、13に比べて残業持続する傾向があります。 </li></ol>メモリータスク現在のプロトコルの主な焦点がキュー-リコールメモリテスト11を使用して調査からの知見に基づいている、した参加者は、各ピクチャのために言語的な合図を提供し、彼らが覚えている画像に関 ​​する詳細を書面で提供するために持っていた。しかし、他の検索タスクを使用することもできます。特に関連して、いわゆるRKパラダイム月31日参加者は、彼らはアイテムを（遭遇したことのエンコーディング（レスポンスを覚えて）から、特定のコンテキストの詳細を思い出すか、単に慣れている印象を持っているかどうかに基づいて取得された各項目を分類したメモリの認識タスクは、 ）応答を知っている。 fMRIのと、感情的な刺激で使用され、このタスクは回想-対精通ベースの検索32だけでなく、検索のこれら2つのタイプの感情の差の影響の同定、および関連神経相関12の神経相関の解離ではないだけが可能になります。 fMRIのデータ解析統計的パラメトリックマッピング（SPM：（ <a href="http://www.fil.ion.uc.uk/spm/">http://www.fil.ion.uc.uk/spm/</a>組み合わせで）社内のMatlabベースのツールは、データ分析のために使用されたとの前処理が一般的に含まれる：。品質保証を、画像アライメント、動きの補正、共同登録、正規化、および平滑化（8 mm 3のカーネル）が、分析は興味の解剖学的に定義された領域（ROI）を含む場合は、（11を参照）データを平滑化し、正規化されていない場合、空間的な特異性が得られる増加した。個人およびグループレベルの統計分析は、その後の私に基づいて符号化/検索脳活動の比較を伴うモリーのパフォーマンス。メモリ関連の活動に対する感情の調節効果は、中性刺激（図2参照）よりも感情的より大きなES / DMまたはRSの活性を示す領域で見ることができます。 MTLの活動をターゲットに捜査が刺激（感情的な対中立メモリ、11）の感情的なコンテンツにリンクされている機能的解離、の識別を可能にする、メモリの種類は、（項目対ソースのメモリ、19）研究され、または検索の種類タスク（回想-対熟知ベース、12、36）。差動感情的な対中立メモリ10、20に関与する前頭前野（PFC）、のMTL特に対象地域の外での脳領域の調査。 <img src="/files/ftp_upload/2433/2433fig2.jpg" alt="図2" /> 図2。エモーション変調の影響を受けやすいメモリ関連の脳活動の識別感情変調（EmoDm / RS&gt; NeuDm / RS、すなわち、 感情 X メモリの相互作用を示す）に敏感なメモリに関連する脳活動の同定できることに加えて、この設計を、まただけでなく、メモリの全体的な効果[（;活動の知覚/一般的な処理[すなわち、 感情の主効果を示す（エモーションR +エモーションF）&gt;（ニュートラルR +ニュートラルF）]を上に感情の影響を反映する脳領域の同定が可能に感情R +ニュートラルR）&gt;（エモーションF +ニュートラルF）、すなわち、 メモリの主な効果を示す]。 4。代表的な結果： 情動記憶の研究におけるSMPの雇用は、感情の記憶増強効果の神経相関に光をあてることに成功証明されている。 変調仮説 37、38の強力なサポートを提供し、メモリの符号化の研究は、感情が記憶に関連する脳領域の活動で、扁桃体、感情処理に関わる脳の領域から調節の影響を介してメモリを拡張するという考えを支持する証拠を発見したような内側側頭葉（MTL 11、図3を参照）と前頭前皮質（PFC 10）など。また、記憶の想起の研究では、変調の仮説も検索に適用される、そしてその様々なMTLの領域は、差動感情的な思い出12の回想-対精通ベースの検索に関与しているというさらなる証拠を提供しています。 <img src="/files/ftp_upload/2433/2433fig3.jpg" alt="図3" /> 図3。変調仮説の証拠が。変調の仮説をサポートする、感情の記憶増強効果が増加したES / DM活動（）および（B）扁桃体とMTLメモリシステム（C＆Dに示す）の間の相互作用に関連していた。棒グラフは、感情的で、中立的なES / DMのパーセントの信号の変化、などのピーク活性化のスライスから抽出し、半球の平均値を比較します。散布図は、ES /扁桃体と感情とニュートラルの画像の嗅内皮質のDM活動との間の共変動を示す。 ES / DM =エンコード成功/メモリによる違い。 Dolcosらから。11は 、許可を得て。

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A., Corkin, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+routes+to+emotional+memory:+distinct+neural+processes+for+valence+and+arousal.">Two routes to emotional memory: distinct neural processes for valence and arousal.</a> Proc Natl Acad Sci U S A. 101, 3310-3315 (2004).</li><li>Kensinger, E. A., Garoff-Eaton, R. J., Schacter, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory+for+specific+visual+details+can+be+enhanced+by+negative+arousing+content.">Memory for specific visual details can be enhanced by negative arousing content.</a> Journal of Memory and Language. 54, 99-112 (2006).</li><li>Bradley, M. M., Cuthbert, B. N., Lang, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Picture+media+and+emotion:+effects+of+a+sustained+affective+context.">Picture media and emotion: effects of a sustained affective context.</a> Psychophysiology. 33, 662-670 (1996).</li><li>Lang, P. J., Bradley, M. M., Cuthbert, B. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technical+Report+A-8.">Technical Report A-8.</a> International affective picture system (IAPS): Affective ratings of pictures and instruction manual. , (2008).</li><li>Lang, P. J., Bradley, M. M., Cuthbert, B. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technical+report+C-1.">Technical report C-1.</a> Affective norms for English workds (ANEW): Stimuli, instruction manual and affective ratings. , (2008).</li><li>Bradley, M. M., Lang, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+emotion:+the+Self-Assessment+Manikin+and+the+Semantic+Differential.">Measuring emotion: the Self-Assessment Manikin and the Semantic Differential.</a> J Behav Ther Exp Psychiatry. 25, 49-59 (1994).</li><li>Kensinger, E. 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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. 54, 1063-1070 (1988).</li><li>Spielberger, C. D., Gorsuch, R. L., Lushene, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Manual for the State-Trait Anxiety Inventory. , (1970).</li><li>Voyvodic, J. 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利害の衝突は宣言されません。

この実験的なプロトコルは、特に一般/知覚処理時の感情の変調の影響を受けやすい脳領域における活動からの感情によって変調を受けやすいメモリに関連する脳活動を分離することによって、感情の記憶増強効果の神経相関の調査が可能になります。この設計は、メモリ上に感情の影響の基礎となる神経機構の理解を推進してきました、そしてその多様性与えられたそれは、SMPは、イベント...

brain imaging investigation of the memory-enhancing effect of emotion

感情的なイベントは、より良い非感情的なイベントの1,2よりも記憶される傾向にある。認知と情動神経科学の一つの目標は、メモリ上で感情のこの増強効果の基礎となる神経機構を理解することです。感情の記憶増強効果の調査で特に影響力のある実証されている方法は、いわゆる後続のメモリパラダイム（SMP）です。このメソッドは当初、非感情的な思い出3の神経相関を調査するために使用され、最近では我々と他の人にも感情的なメモリ（5-7 4に概説）の研究に正常に適用されました。ここで、我々は、イベントに関連する機能的磁気共鳴画像（fMRI）と一緒にSMPを使用して、感情の記憶増強効果の神経相関の調査を可能にするプロトコルを記述する。 SMPの重要な特徴は、それが具体的知覚に関連するより一般的な活動からメモリに関連する脳活動の分離を可能にすることです。また、感情的な刺激の影響を調査するのコンテキスト内で、SMPは、アクティビティの一般的な/知覚とメモリ固有の処理の両方の感情的な変調を受けやすい脳領域の同定を可能にします。このプロトコルは、だけでなく、うつ病や心的外傷後ストレス障害（PTSDで苦しむ人々のような感情的なイベントを、覚えておくの感情の認知とバイアスの神経相関の変化があると臨床患者において、健常8-15に使用することができます。 ）16、17。

Watch this Scientific Journal Video about Brain Imaging Investigation of the Memory-Enhancing Effect of Emotion at JoVE.com

Brain Imaging Investigation of the Memory-Enhancing Effect of Emotion

私たちは、感情の記憶増強効果の神経相関を調査する機能的磁気共鳴イメージングを使用するプロトコルを示す。このプロトコルは、脳活動の識別は特に、より一般的な知覚処理に反して、メモリ関連の処理、にリンクできるよう、そして健康と臨床集団で使用することができます。

感情の記憶増強効果の脳イメージング研究

Emotional events tend to be better remembered than non-emotional events1,2. One goal of cognitive and affective neuroscientists is 
to understand the ...

brain-imaging-investigation-of-the-memory-enhancing-effect-of-emotion

Research

JoVE Journal

Neuroscience

16.2K ビュー.  University of Alberta. 私たちは、感情の記憶増強効果の神経相関を調査する機能的磁気共鳴イメージングを使用するプロトコルを示す。このプロトコルは、脳活動の識別は特に、より一般的な知覚処理に反して、メモリ関連の処理、にリンクできるよう、そして健康と臨床集団で使用することができます。

ビデオ: Brain Imaging Investigation of the Memory-Enhancing Effect of Emotion

Brain Imaging Investigation of the Neural Correlates of Observing Virtual Social Interactions

The ability to gauge social interactions is crucial in the assessment of others&#x2019; intentions. Factors such as facial expressions and body language affect our decisions in personal and professional life alike 1. These "friend or foe" judgements are often based on first impressions, which in turn may affect our decisions to "approach or avoid". Previous studies investigating the neural correlates of social cognition tended to use static facial stimuli 2. Here, we illustrate an experimental design in which whole-body animated characters were used in conjunction with functional magnetic resonance imaging (fMRI) recordings. Fifteen participants were presented with short movie-clips of guest-host interactions in a business setting, while fMRI data were recorded; at the end of each movie, participants also provided ratings of the host behaviour. This design mimics more closely real-life situations, and hence may contribute to better understanding of the neural mechanisms of social interactions in healthy behaviour, and to gaining insight into possible causes of deficits in social behaviour in such clinical conditions as social anxiety and autism 3.

This article demonstrates an experimental design in which whole-body animated characters are used in conjunction with functional magnetic resonance imaging (fMRI) to investigate the neural correlates of observing virtual social interactions.

仮想社会的相互作用を観察するの神経相関の脳イメージング研究

The ability to gauge social interactions is crucial in the assessment of others&#x2019; intentions. Factors such as facial expressions and body language ...

神経科学

Brain Imaging Investigation of the Neural Correlates of Emotional Autobiographical Recollection

Recollection of emotional autobiographical memories (AMs) is important to healthy cognitive and affective functioning 1 - remembering positive AMs is associated with increased personal well-being and self-esteem 2, whereas remembering and ruminating on negative AMs may lead to affective disorders 3. Although significant progress has been made in understanding the brain mechanisms underlying AM retrieval in general (reviewed in 4, 5), less is known about the effect of emotion on the subjective re-experience of AMs and the associated neural correlates. This is in part due to the fact that, unlike the investigations of the emotion effect on memory for laboratory-based microevents (reviewed in 6, 7-9), often times AM studies do not have a clear focus on the emotional aspects of remembering personal events (but see 10). Here, we present a protocol that allows investigation of the neural correlates of recollecting emotional AMs using functional magnetic resonance imaging (fMRI). Cues for these memories are collected prior to scanning by means of an autobiographical memory questionnaire (AMQ), therefore allowing for proper selection of emotional AMs based on their phenomenological properties (i.e., intensity, vividness, personal significance). This protocol can be used in healthy and clinical populations alike.

We present a protocol that allows investigation of the neural correlates of recollecting emotional autobiographical memories, using functional magnetic resonance imaging. This protocol can be used with both healthy and clinical participants.

感情的な自伝追憶の神経相関の脳イメージング研究

Recollection of emotional autobiographical memories (AMs) is important to healthy cognitive and affective functioning 1 - remembering positive AMs is ...

Brain Imaging Investigation of the Neural Correlates of Emotion Regulation

The ability to control/regulate emotions is an important coping mechanism in the face of emotionally stressful situations. Although significant progress has been made in understanding conscious/deliberate emotion regulation (ER), less is known about non-conscious/automatic ER and the associated neural correlates. This is in part due to the problems inherent in the unitary concepts of automatic and conscious processing1. Here, we present a protocol that allows investigation of the neural correlates of both deliberate and automatic ER using functional magnetic resonance imaging (fMRI). This protocol allows new avenues of inquiry into various aspects of ER. For instance, the experimental design allows manipulation of the goal to regulate emotion (conscious vs. non-conscious), as well as the intensity of the emotional challenge (high vs. low). Moreover, it allows investigation of both immediate (emotion perception) and long-term effects (emotional memory) of ER strategies on emotion processing. Therefore, this protocol may contribute to better understanding of the neural mechanisms of emotion regulation in healthy behaviour, and to gaining insight into possible causes of deficits in depression and anxiety disorders in which emotion dysregulation is often among the core debilitating features.

We present a protocol that allows investigation of the neural correlates of deliberate and automatic emotion regulation, using functional magnetic resonance imaging. This protocol can be used in healthy participants, both young and older, as well as in clinical patients.

情動表出の制御の神経相関の脳イメージング研究

The ability to control/regulate emotions is an important coping mechanism in the face of emotionally stressful situations. Although significant progress ...

Brain Imaging Investigation of the Impairing Effect of Emotion on Cognition

Emotions can impact cognition by exerting both enhancing (e.g., better memory for emotional events) and impairing (e.g., increased emotional distractibility) effects (reviewed in 1). Complementing our recent protocol 2 describing a method that allows investigation of the neural correlates of the memory-enhancing effect of emotion (see also 1, 3-5), here we present a protocol that allows investigation of the neural correlates of the detrimental impact of emotion on cognition. The main feature of this method is that it allows identification of reciprocal modulations between activity in a ventral neural system, involved in 'hot' emotion processing (HotEmo system), and a dorsal system, involved in higher-level 'cold' cognitive/executive processing (ColdEx system), which are linked to cognitive performance and to individual variations in behavior (reviewed in 1). Since its initial introduction 6, this design has proven particularly versatile and influential in the elucidation of various aspects concerning the neural correlates of the detrimental impact of emotional distraction on cognition, with a focus on working memory (WM), and of coping with such distraction 7,11, in both healthy 8-11 and clinical participants 12-14.

We present a protocol that allows investigation of the neural mechanisms mediating the detrimental impact of emotion on cognition, using functional magnetic resonance imaging. This protocol can be used with both healthy and clinical participants.

認知に関する情動の損なう効果の脳イメージング研究

Emotions can impact cognition by exerting both enhancing (e.g., better memory for emotional events) and impairing (e.g., increased emotional ...

Live Imaging of Mitosis in the Developing Mouse Embryonic Cortex

Although of short duration, mitosis is a complex and dynamic multi-step process fundamental for development of organs including the brain. In the developing cerebral cortex, abnormal mitosis of neural progenitors can cause defects in brain size and function. Hence, there is a critical need for tools to understand the mechanisms of neural progenitor mitosis. Cortical development in rodents is an outstanding model for studying this process. Neural progenitor mitosis is commonly examined in fixed brain sections. This protocol will describe in detail an approach for live imaging of mitosis in ex vivo embryonic brain slices. We will describe the critical steps for this procedure, which include: brain extraction, brain embedding, vibratome sectioning of brain slices, staining and culturing of slices, and time-lapse imaging. We will then demonstrate and describe in detail how to perform post-acquisition analysis of mitosis. We include representative results from this assay using the vital dye Syto11, transgenic mice (histone H2B-EGFP and centrin-EGFP), and in utero electroporation (mCherry-&#945;-tubulin). We will discuss how this procedure can be best optimized and how it can be modified for study of genetic regulation of mitosis. Live imaging of mitosis in brain slices is a flexible approach to assess the impact of age, anatomy, and genetic perturbation in a controlled environment, and to generate a large amount of data with high temporal and spatial resolution. Hence this protocol will complement existing tools for analysis of neural progenitor mitosis.

Neural progenitor mitosis is a critical parameter of neurogenesis. Much of our understanding of neural progenitor mitosis is based on analysis of fixed tissue. Live imaging in embryonic brain slices is a versatile technique to assess mitosis with high temporal and spatial resolution in a controlled environment.

発生中のマウス胚性皮質における有糸分裂のライブイメージング

Although of short duration, mitosis is a complex and dynamic multi-step process fundamental for development of organs including the brain. In the ...

Live Imaging of the Ependymal Cilia in the Lateral Ventricles of the Mouse Brain

Multiciliated ependymal cells line the ventricles in the adult brain. Abnormal function or structure of ependymal cilia is associated with various neurological deficits. The current ex vivo live imaging of motile ependymal cilia technique allows for a detailed study of ciliary dynamics following several steps. These steps include: mice euthanasia with carbon dioxide according to protocols of The University of Toledo&#8217;s Institutional Animal Care and Use Committee (IACUC); craniectomy followed by brain removal and sagittal brain dissection with a vibratome or sharp blade to obtain very thin sections through the brain lateral ventricles, where the ependymal cilia can be visualized. Incubation of the brain&#8217;s slices in a customized glass-bottom plate containing Dulbecco&#8217;s Modified Eagle&#8217;s Medium (DMEM)/High-Glucose at 37 &#176;C in the presence of 95%/5% O2/CO2 mixture is essential to keep the tissue alive during the experiment. A video of the cilia beating is then recorded using a high-resolution differential interference contrast microscope. The video is then analyzed frame by frame to calculate the ciliary beating frequency. This allows distinct classification of the ependymal cells into three categories or types based on their ciliary beating frequency and angle. Furthermore, this technique allows the use of high-speed fluorescence imaging analysis to characterize the unique intracellular calcium oscillation properties of ependymal cells as well as the effect of pharmacological agents on the calcium oscillations and the ciliary beating frequency. In addition, this technique is suitable for immunofluorescence imaging for ciliary structure and ciliary protein localization studies. This is particularly important in disease diagnosis and phenotype studies. The main limitation of the technique is attributed to the decrease in live motile cilia movement as the brain tissue starts to die.

Using high-resolution differential interference contrast (DIC) microscopy, an ex vivo observation of the beating of motile ependymal cilia located within the mouse brain ventricles is demonstrated by live-imaging. The technique allows a recording of the unique ciliary beating frequency and beating angle as well as their intracellular calcium oscillation pacing properties.

マウスの脳の側脳室における上衣繊毛のライブイメージング

Multiciliated ependymal cells line the ventricles in the adult brain. Abnormal function or structure of ependymal cilia is associated with various ...

Effects of Transcranial Alternating Current Stimulation on the Primary Motor Cortex by Online Combined Approach with Transcranial Magnetic Stimulation

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific frequency and in turn modulate ongoing cortical oscillatory activity. This neurotool allows the establishment of a causal link between endogenous oscillatory activity and behavior. Most of the tACS studies have shown online effects of tACS. However, little is known about the underlying action mechanisms of this technique because of the AC-induced artifacts on Electroencephalography (EEG) signals. Here we show a unique approach to investigate online physiological frequency-specific effects of tACS of the primary motor cortex (M1) by using single pulse Transcranial Magnetic Stimulation (TMS) to probe cortical excitability changes. In our setup, the TMS coil is placed over the tACS electrode while Motor Evoked Potentials (MEPs) are collected to test the effects of the ongoing M1-tACS. So far, this approach has mainly been used to study the visual and motor systems. However, the current tACS-TMS setup can pave the way for future investigations of cognitive functions. Therefore, we provide a step-by-step manual and video guidelines for the procedure.

Transcranial Alternating Current Stimulation (tACS) allows the modulation of cortical excitability in a frequency-specific fashion. Here we show a unique approach which combines online tACS with single pulse Transcranial Magnetic Stimulation (TMS) in order to "probe" cortical excitability by means of Motor Evoked Potentials.

経頭蓋交流電流刺激オンライン結合による一次運動野の経頭蓋磁気刺激の効果

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific ...

Large-scale Three-dimensional Imaging of Cellular Organization in the Mouse Neocortex

The mammalian neocortex is composed of many types of excitatory and inhibitory neurons, each with specific electrophysiological and biochemical properties, synaptic connections, and in vivo functions, but their basic functional and anatomical organization from cellular to network scale is poorly understood. Here we describe a method for the three-dimensional imaging of fluorescently-labeled neurons across large areas of the brain for the investigation of the cortical cellular organization. Specific types of neurons are labeled by the injection of fluorescent retrograde neuronal tracers or expression of fluorescent proteins in transgenic mice. Block brain samples, e.g., a hemisphere, are prepared after fixation, made transparent with tissue clearing methods, and subjected to fluorescent immunolabeling of the specific cell types. Large areas are scanned using confocal or two-photon microscopes equipped with large working distance objectives and motorized stages. This method can resolve the periodic organization of the cell type-specific microcolumn functional modules in the mouse neocortex. The procedure can be useful for the study of three-dimensional cellular architecture in the diverse brain areas and other complex tissues.

Here we describe a procedure for tissue clearing, fluorescent labeling, and large-scale imaging of mouse brain tissue which, thereby, enables visualization of the three-dimensional organization of cell types in the neocortex.

マウス大脳皮質における細胞組織の大規模三次元イメージング

The mammalian neocortex is composed of many types of excitatory and inhibitory neurons, each with specific electrophysiological and biochemical ...

Advanced Diffusion Imaging in The Hippocampus of Rats with Mild Traumatic Brain Injury

Mild traumatic brain injury (mTBI) is the most common type of acquired brain injury. Since patients with traumatic brain injury show a tremendous variability and heterogeneity (age, gender, type of trauma, other possible pathologies, etc.), animal models play a key role in unraveling factors that are limitations in clinical research. They provide a standardized and controlled setting to investigate the biological mechanisms of injury and repair following TBI. However, not all animal models mimic the diffuse and subtle nature of mTBI effectively. For example, the commonly used controlled cortical impact (CCI) and lateral fluid percussion injury (LFPI) models make use of a craniotomy to expose the brain and induce widespread focal trauma, which are not commonly seen in mTBI. Therefore, these experimental models are not valid to mimic mTBI. Thus, an appropriate model should be used to investigate mTBI. The Marmarou weight drop model for rats induces similar microstructural alterations and cognitive impairments as seen in patients who sustain mild trauma; therefore, this model was selected for this protocol. Conventional computed tomography and magnetic resonance imaging (MRI) scans commonly show no damage following a mild injury, because mTBI induces often only subtle and diffuse injuries. With diffusion weighted MRI, it is possible to investigate microstructural properties of brain tissue, which can provide more insight into the microscopic alterations following mild trauma. Therefore, the goal of this study is to obtain quantitative information of a selected region-of-interest (i.e., hippocampus) to follow up disease progression after obtaining a mild and diffuse brain injury.

The overall goal of this procedure is to obtain quantitative microstructural information of the hippocampus in a rat with mild traumatic brain injury. This is done using an advanced diffusion-weighted magnetic resonance imaging protocol and region-of-interest based analysis of parametric diffusion maps.

軽度の外傷性脳損傷を有するラットの海馬における高度な拡散イメージング

Mild traumatic brain injury (mTBI) is the most common type of acquired brain injury. Since patients with traumatic brain injury show a tremendous ...

Investigation of Spatial Interaction Between Astrocytes and Neurons in Cleared Brains

Combining viral vector transduction and tissue clearing using the CLARITY method makes it possible to simultaneously investigate several types of brain cells and their interactions. Viral vector transduction enables the marking of diverse cell types in different fluorescence colors within the same tissue. Cells can be identified genetically by activity or projection. Using a modified CLARITY protocol, the potential sample size of astrocytes and neurons has grown by 2-3 orders of magnitude. The use of CLARITY allows the imaging of complete astrocytes, which are too large to fit in their entirety in slices, and the examination of the somata with all their processes. In addition, it provides the opportunity to investigate the spatial interaction between astrocytes and different neuronal cell types, namely, the number of pyramidal neurons in each astrocytic domain or the proximity between astrocytes and specific inhibitory neuron populations. This paper describes, in detail, how these methods are to be applied.

Combining viral vector transduction and brain clearing using the CLARITY method allows the investigation of a large number of neurons and astrocytes simultaneously.

クリア脳におけるアストロサイトとニューロンの空間的相互作用の解明

Combining viral vector transduction and tissue clearing using the CLARITY method makes it possible to simultaneously investigate several types of brain ...