我々は彼らのサポートのために、次の感謝は次のとおりです。連邦奨学金委員会が英国の資金調達（HP）、EPSRC（HP）、BBSRC（MFN）と欧州連合（EU）マリー·キュリー·アクション（MFN）。

<ol>
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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+constants+of+h+current+in+layer+II+stellate+cells+differ+along+the+dorsal+to+ventral+axis+of+medial+entorhinal+cortex.">Time constants of h current in layer II stellate cells differ along the dorsal to ventral axis of medial entorhinal cortex.</a> Journal of Neuroscience. 28, 9414-9425 (2008).</li><li>Burgalossi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microcircuits+of+functionally+identified+neurons+in+the+rat+medial+entorhinal+cortex.">Microcircuits of functionally identified neurons in the rat medial entorhinal cortex.</a> Neuron. 70, 773-786 (2011).</li><li>Dodson, P. D., Pastoll, H., Nolan, M. 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開示するものがありません。

我々はここで詳細にMECの背腹軸の範囲を維持する矢状スライス標本を製造するための手順を記載している背腹組織に従うMEC回路特性の調査を容易にする。 重要なステップ 動物から脳を削除します 。脳に作用する圧力を避けるために、特に注意してください。これは、脳の急速な除去よりも重要である。 スライス。スライスがしっかりと保持室の床に接着する必要があります。ビブラトームは、z軸の振動&lt;2μmのを持つ必要があり、高振幅、低速度の設定に使用する必要があります。最適な設定はモデル間で異なることがあり、試行錯誤によって確立されている必要があります。 溶液の温度を監視します 。我々は、スライスの質が切断中と後の温度に敏感であることがわかります。 Outsid<html> eが推奨温度は、健康な神経細胞の数が削減され、ニューロンはシナプス入力にパッチと応答がより困難になる可能性が減少することができます範囲である。 フィールド虹彩絞りのイメージング。ケーラー照明とセンタリングは、録音前の細胞の可視化のために重要である。 トラブルシューティング スライスは、少数の健康な細胞が含まれています。ソリューションを交換してください。ビブラトーム上のz軸の変位を確認してください。溶液の温度を確認してください。切断スライスの向きを確認してください。 スライス内のニューロンは、参照することが困難である。ケーラー照明が正しく設定されていることを確認。顕微鏡レンズが汚れていない確認してください。 gigasealsを形成する難しさ。記録電極の形状を確認してください。細胞死は、例えば円形細胞の形状や、コントラストの高い細胞の兆候スライスを調べます。 スライスの ve_content &quot;&gt; 画像を整列することが困難である。スライスが最大の詳細をキャプチャするためにフォーカスされていることを確認します。自動画像調整&gt; 40％の画像の重なりのために通常は十分です。 制限事項矢状スライス標本の1つの潜在的な制限は、MEC内の非常に長い距離シナプスの接続が保持されない場合がありますので、矢状面9、15に直接並列実行し、ないかもしれないということです。これらの接続を維持することはどちら吻側 - 尾側平面の角度でビブラトーム上での脳半球をhemisectingまたはマウントすることで、カットスライスの角度を変更するには、準備手順を変更する必要があります。同様の考慮事項は、内側中隔からの例の求心性入力の場合、他の接続の保全を申請することができます。 水平MECスライス標本との比較<ol><li>背腹軸の位置の正確な測定が重要である背腹軸および生理学的特性に関する沿った位置との間の量的関係を確立するため。水平方向のスライス標本7,8,11,12では、カットの正確な背腹軸の位置を決定することは困難であり、これは一貫した基準位置を確立する複雑になります。対照的に、矢状スライス標本でMECの背側の境界線が基準点とスライスのニューロンの深さの知識は、その背腹軸の位置を測定する必要はありませとして確立するのは簡単です。矢スライスしたがって、非常に背腹軸の位置と、携帯電話や回路の特性との間の量的関係をより迅速かつ正確な調査を可能にする、背腹軸の位置の正確な測定を容易にします。 </li><li>分子勾配はMECの地形回路組織のために重要であり、蛍光または他の標識技術を用いて可視化することができます。水平スライスと比較して、その目で電子信号が別の背腹軸の位置でスライス間で比較されなければならない、矢準備が背腹軸の分子勾配を簡単に、よりきめの細かい、より信頼性の高い可視化および定量化が可能になります。 </li><li> 。シナプスの接続に背腹軸勾配はMEC機能に重要な役割を果たす可能性があります。矢状スライス標本では、接続内および異なる場所の間にMECの背腹軸に沿って、さまざまな背腹で均等に回路の整合性を維持するためにどのように異なるか調査するために重要であるMEC内背側と腹側の領域の間のシナプス結合を保持することができます場所。接続が背腹軸に沿って実質的に変化する場合、水平方向のスライスは同じようにそれぞれの極端な全体の機能的なマイクロを含む可能性が低くなります。 </li></ol>将来の開発とアプリケーション分子決定因子の知識MECの細胞のアイデンティティのニューロンのアイデンティティの決定的な特性を有効にして、背腹軸の位置のより正確な決定を許可します。 矢状スライス標本を用いて、複数のニューロンの細胞型および位置特異的活性の地形主催応答は1つのスライス内（電圧感受性色素やカルシウムイメージングを用いた例）を同時に観察することができた。 optogeneticツールとの組み合わせで、矢状製剤は、背腹軸に沿った異なる位置で選択的活性化を可能にします。別の背腹軸の位置で超小型回路が空間的に標的と活性化にどのように応答するかを尋ねると、MECマイクロ回路の機能に重要な洞察を提供することができます。 我々は、この準備は生理学的特性に背腹軸勾配を調査するための、シンプル、堅牢で標準化された基盤を提供し、促進することを期待してこれらの勾配がMECの情報処理特性に寄与する方法を理解する。

<table border="1"><tbody><tr><td></td><td> ACSF（mm）を切断 </td><td> 標準ACSF（MM） </td><td> 内部液（mM）を </td><td> CASNumber </td><td> サプライヤのカタログ番号 </td></tr><tr><td> NaClを</td><td> 86 </td><td> 124 </td><td></td><td> 7647-14-5 </td><td>シグマS9888 </td></tr><tr><td>のNaH 2 PO 4 </td><td> 1.2 </td><td> 1.2 </td><td></td><td> 13472-35-0 </td><td>シグマ71505 </td></tr><tr><td>塩化カリウム</td><td> 2.5 </td><td> 2.5 </td><td> 10 </td><td> 7447-40-7 </td><td>シグマP3911 </td></tr><tr><td> NaHCO 3を </td><td> 25 </td><td> 25 </td><td></td><td> 144-55-8 </td><td>フィッシャーS/4240 </td></tr><tr><td>グルコース</td><td> 25 </td><td> 20 </td><td></td><td> 50-99-7 </td><td>シグマG5767 </td></tr><tr><td>スクロース</td><td> 75 </td><td></td><td></td><td> 57-50-1 </td><td>シグマS5016 </td></tr><tr><td> CACl 2の </td><td> 0.5 </td><td> 2 </td><td></td><td> 10043-52-4 </td><td> VWR 190464K </td></tr><tr><td> MgCl 2を </td><td> 7 </td><td> 1 </td><td> 2 </td><td> 7786-30-3 </td><td>シグマ63020 </td></tr><tr><td> Kグルコン酸</td><td></td><td></td><td> 130 </td><td> 299-27-4 </td><td>シグマG4500 </td></tr><tr><td> HEPES </td><td></td><td></td><td> 10 </td><td> 7365-45-9 </td><td>シグマH3375 </td></tr><tr><td> EGTA </td><td></td><td></td><td> 0.1 </td><td> 67-42-5 </td><td>シグマE4378 </td></tr><tr><td>のNa 2 ATP </td><td></td><td></td><td> 2 </td><td> 34369-07-8 </td><td>シグマA7699 </td></tr><tr><td>のNa 2 GTP </td><td></td><td></td><td> 0.3 </td><td> 36051-31-7 </td><td>シグマG8877 </td></tr><tr><td> NaPhospho-クレアチン</td><td></td><td></td><td> 10 </td><td> 19333-65-4 </td><td>シグマP7936 </td></tr><tr><td>ビオシチン（オプション） </td><td></td><td></td><td> 2.7 </td><td> 576-19-2 </td><td&gt;シグマB4261 </td></tr></tbody></table> 表1。切削ACSF、標準ACSFとK-グルコン内部ソリューションのレシピ。

preparation of parasagittal slices for the investigation of dorsal-ventral organization of the rodent medial entorhinal cortex

脳内の計算は、シナプス入力に適切に応答ニューロンに依存しています。ニューロンは、それらの補数、彼らはシナプス入力に応答する方法を決定する膜のイオンチャネルの分布が異なります。しかし、動物の動作でこれらの細胞の特性と神経機能の関係はよく理解されていません。この問題に対する一つのアプローチは、それらがコード情報、または計算上の個々のニューロンのマップの位置は、彼らが1を実施している地形的に編成神経回路を検討することである。このアプローチを用いた実験は、感覚や認知回路2,3内の情報の符号化の基礎となるシナプス応答のチューニングのための原則を示唆している。 内側嗅内皮質（MEC）の背腹軸に沿って空間的な表現の地形組織は、細胞のメカニズムと計算の間の関係を確立する機会を提供していますI空間認知のためにmportant。げっ歯類のMECのレイヤーIIのニューロンは、格子状のフィールド4-6焼成を使用して場所をエンコードします。次第に腹の位置でニューロンは、この距離より大きい1メートルに増加し、一方MECの背側の位置で見つかった神経細胞のグリッドを形成する個々の焼成のフィールドの間の距離は、30cmのオーダーである。いくつかの研究は、これらの細胞の特性は、空間計算2,7-10に重要であることが示唆され、グリッド発射フィールド間の間隔と同様、彼らの背腹の位置に応じて異なることMECのレイヤーIIのニューロンの細胞の性質を明らかにした。 ここでは、MECニューロンの生物物理学と解剖学的特性の地形組織のMEC可能にする調査の背腹軸の範囲を維持する脳スライスからの調製および電気生理学的記録のための手順について説明します。識別さnの背腹軸の位置それは、スライスの正確な背腹の位置の基準点を確立することは困難であるとして、解剖学的ランドマークの相対euronsは、MEC 7,8,11,12の水平スライスを使用するプロトコルを正確に確立することは困難である。我々は記述の手順では、MECと同様に、分子勾配2,10の可視化の背腹軸に沿って記録された細胞の位置を正確かつ一貫性のある測定を可能にします。手順は、成体マウスで使用するために開発されている（&gt; 28日間）に成功し、最大1.5歳までのマウスで採用されている。調整に彼らは若いマウスまたは他の齧歯類の種と一緒に使用することができます。調製と測定の標準化されたシステムは、この領域の細胞とマイクロ回路の特性の体系的な調査を支援します。

Watch this Scientific Journal Video about Preparation of Parasagittal Slices for the Investigation of Dorsal-ventral Organization of the Rodent Medial Entorhinal Cortex at JoVE.com

Preparation of Parasagittal Slices for the Investigation of Dorsal-ventral Organization of the Rodent Medial Entorhinal Cortex

我々は内側嗅内皮質（MEC）の背腹軸を維持する脳スライスからの調製および電気生理学的記録のための手順について説明します。場所の神経エンコーディングはMEC内の背腹軸の組織を次のようなので、これらの手順は、ナビゲーションやメモリのための重要な細胞メカニズムの調査を容易にします。

げっ歯類の内側嗅内皮質の背腹軸機構の調査のための矢状スライスの作成

Computation in the brain relies on neurons responding appropriately to their synaptic inputs. Neurons differ in their complement and distribution of ...

preparation-parasagittal-slices-for-investigation-dorsal-ventral

Nanyang Technological University

Research

JoVE Journal

Neuroscience

15.6K ビュー.  University of Edinburgh. 我々は内側嗅内皮質（MEC）の背腹軸を維持する脳スライスからの調製および電気生理学的記録のための手順について説明します。場所の神経エンコーディングはMEC内の背腹軸の組織を次のようなので、これらの手順は、ナビゲーションやメモリのための重要な細胞メカニズムの調査を容易にします。

ビデオ: Preparation of Parasagittal Slices for the Investigation of Dorsal-ventral Organization of the Rodent Medial Entorhinal Cortex

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 ...

神経科学

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 Memory-Enhancing Effect of Emotion

Emotional events tend to be better remembered than non-emotional events1,2. One goal of cognitive and affective neuroscientists is 
to understand the neural mechanisms underlying this enhancing effect of emotion on memory. A method that has proven particularly influential in the 
investigation of the memory-enhancing effect of emotion is the so-called subsequent memory paradigm (SMP). This method was originally used to investigate the 
neural correlates of non-emotional memories3, and more recently we and others also applied it successfully to studies of emotional memory (reviewed in4, 5-7). 
Here, we describe a protocol that allows investigation of the neural correlates of the memory-enhancing effect of emotion using the SMP in conjunction with 
event-related functional magnetic resonance imaging (fMRI). An important feature of the SMP is that it allows separation of brain activity specifically 
associated with memory from more general activity associated with perception. Moreover, in the context of investigating the impact of emotional stimuli, 
SMP allows identification of brain regions whose activity is susceptible to emotional modulation of both general/perceptual and memory-specific processing. 
This protocol can be used in healthy subjects8-15, as well as in clinical patients where there are alterations in the neural correlates of emotion perception 
and biases in remembering emotional events, such as those suffering from depression and post-traumatic stress disorder (PTSD)16, 17.

We present a protocol that uses functional magnetic resonance imaging to investigate the neural correlates of the memory-enhancing effect of emotion. This protocol allows identification of brain activity specifically linked to memory-related processing, contrary to more general perceptual processing, and can be used with healthy and clinical populations.

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 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 ...

Preparation of Acute Subventricular Zone Slices for Calcium Imaging

The subventricular zone (SVZ) is one of the two neurogenic zones in the postnatal brain. The SVZ contains densely packed cells, including neural progenitor cells with astrocytic features (called SVZ astrocytes), neuroblasts, and intermediate progenitor cells. Neuroblasts born in the SVZ tangentially migrate a great distance to the olfactory bulb, where they differentiate into interneurons. Intercellular signaling through adhesion molecules and diffusible signals play important roles in controlling neurogenesis. Many of these signals trigger intercellular calcium activity that transmits information inside and between cells. Calcium activity is thus reflective of the activity of extracellular signals and is an optimal way to understand functional intercellular signaling among SVZ cells.
Calcium activity has been studied in many other regions and cell types, including mature astrocytes and neurons. However, the traditional method to load cells with calcium indicator dye (i.e. bath loading) was not efficient at loading all SVZ cell types. Indeed, the cellular density in the SVZ precludes dye diffusion inside the tissue. In addition, preparing sagittal slices will better preserve the three-dimensional arrangement of SVZ cells, particularly the stream of neuroblast migration on the rostral-caudal axis.
Here, we describe methods to prepare sagittal sections containing the SVZ, the loading of SVZ cells with calcium indicator dye, and the acquisition of calcium activity with time-lapse movies. We used Fluo-4 AM dye for loading SVZ astrocytes using pressure application inside the tissue. Calcium activity was recorded using a scanning confocal microscope allowing a precise resolution for distinguishing individual cells. Our approach is applicable to other neurogenic zones including the adult hippocampal subgranular zone and embryonic neurogenic zones. In addition, other types of dyes can be applied using the described method.

A method to load subventricular zone (SVZ) cells with calcium indicator dyes for recording calcium activity is described. The postnatal SVZ contains tightly packed cells including neural progenitor cells and neuroblasts. Rather than using bath loading we injected the dye by pressure inside the tissue allowing better dye diffusion.

カルシウムイメージングの急性脳室下帯スライスの調製

The subventricular zone (SVZ) is one of the two neurogenic zones in the postnatal brain. The SVZ contains densely packed cells, including neural ...

A Comprehensive Protocol for Manual Segmentation of the Medial Temporal Lobe Structures

The present paper describes a comprehensive protocol for manual tracing of the set of brain regions comprising the medial temporal lobe (MTL): amygdala, hippocampus, and the associated parahippocampal regions (perirhinal, entorhinal,&#160;and parahippocampal proper). Unlike most other tracing protocols available, typically focusing on certain MTL areas (e.g., amygdala and/or hippocampus), the integrative perspective adopted by the present tracing guidelines allows for clear localization of all MTL subregions. By integrating information from a variety of sources, including extant tracing protocols separately targeting various MTL structures, histological reports, and brain atlases, and with the complement of illustrative visual materials, the present protocol provides an accurate, intuitive, and convenient guide for understanding the MTL anatomy. The need for such tracing guidelines is also emphasized by illustrating possible differences between automatic and manual segmentation protocols. This knowledge can be applied toward research involving not only structural MRI investigations but also structural-functional colocalization and fMRI signal extraction from anatomically defined ROIs, in healthy and clinical groups alike.

The present work provides a comprehensive set of guidelines for manually tracing the medial temporal lobe (MTL) structures. This protocol can be applied to research involving structural and/or combined structural-functional magnetic resonance imaging (MRI) investigations of the MTL, in both healthy and clinical groups.

内側側頭葉構造の手動分割のための総合的なプロトコル

The present paper describes a comprehensive protocol for manual tracing of the set of brain regions comprising the medial temporal lobe (MTL): amygdala, ...

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method

This protocol is a practical guide to the N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. Numerous recent studies have validated the utility of this method for enhancing neuronal preservation and overall brain slice viability. The implementation of this technique by early adopters has facilitated detailed investigations into brain function using diverse experimental applications and spanning a wide range of animal ages, brain regions, and cell types. Steps are outlined for carrying out the protective recovery brain slice technique using an optimized NMDG artificial cerebrospinal fluid (aCSF) media formulation and enhanced procedure to reliably obtain healthy brain slices for patch clamp electrophysiology. With this updated approach, a substantial improvement is observed in the speed and reliability of gigaohm seal formation during targeted patch clamp recording experiments while maintaining excellent neuronal preservation, thereby facilitating challenging experimental applications. Representative results are provided from multi-neuron patch clamp recording experiments to assay synaptic connectivity in neocortical brain slices prepared from young adult transgenic mice and mature adult human neurosurgical specimens. Furthermore, the optimized NMDG protective recovery method of brain slicing is compatible with both juvenile and adult animals, thus resolving a limitation of the original methodology. In summary, a single media formulation and brain slicing procedure can be implemented across various species and ages to achieve excellent viability and tissue preservation.

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method

This protocol demonstrates the implementation of an optimized N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. A single media formulation is used to reliably obtain healthy brain slices from animals of any age and for diverse experimental applications.

最適化されたNを使用して急性脳スライスの準備-メチル-D-glucamine 保護回復法

This protocol is a practical guide to the N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. Numerous recent studies have ...

The Preparation of Oblique Spinal Cord Slices for Ventral Root Stimulation

Electrophysiological recordings from spinal cord slices have proven to be a valuable technique to investigate a wide range of questions, from cellular to network properties. We show how to prepare viable oblique slices of the spinal cord of young mice (P2 - P11). In this preparation, the motoneurons retain&#160;their axons coming out from the ventral roots of the&#160;spinal cord. Stimulation of these axons elicits back-propagating action potentials invading the motoneuron&#160;somas and exciting the motoneuron&#160;collaterals within the spinal cord. Recording of antidromic action potentials is an immediate, definitive and elegant way to characterize motoneuron identity, which surpasses&#160;other identification methods. Furthermore, stimulating the motoneuron collaterals is a simple and reliable way to excite the collateral targets of the motoneurons within the spinal cord, such as other motoneurons or Renshaw cells. In this protocol, we present antidromic recordings from the motoneuron somas as well as Renshaw cell excitation, resulting from ventral root stimulation.

We show how to prepare oblique slices of the spinal cord in young mice. This preparation allows for the stimulation of the ventral roots.

前根刺激のための斜め脊髄スライスの調製

Electrophysiological recordings from spinal cord slices have proven to be a valuable technique to investigate a wide range of questions, from cellular to ...

Preparation of Horizontal Slices of Adult Mouse Retina for Electrophysiological Studies

Vertical slice preparations are well established to study circuitry and signal transmission in the adult mammalian retina. The plane of sectioning in these preparations is perpendicular to the retinal surface, making it ideal for the study of radially oriented neurons like photoreceptors and bipolar cells. However, the large dendritic arbors of horizontal cells, wide-field amacrine cells, and ganglion cells are mostly truncated, leaving markedly reduced synaptic activity in these cells. Whereas ganglion cells and displaced amacrine cells can be studied in a whole-mounted preparation of the retina, horizontal cells and amacrine cells located in the inner nuclear layer are only poorly accessible for electrodes in whole retina tissue.
To achieve maximum accessibility and synaptic integrity, we developed a horizontal slice preparation of the mouse retina, and studied signal transmission at the synapse between photoreceptors and horizontal cells. Horizontal sectioning allows&#160;(1) easy and unambiguous visual identification of horizontal cell bodies for electrode targeting, and (2) preservation of the extended horizontal cell dendritic fields, as a prerequisite for intact and functional cone synaptic input to horizontal cell dendrites.
Horizontal cells from horizontal slices exhibited tonic synaptic activity in the dark, and they responded to brief flashes of light with a reduction of inward current and diminished synaptic activity. Immunocytochemical evidence indicates that almost all cones within the dendritic field of a horizontal cell establish synapses with its peripheral dendrites. The horizontal slice preparation is therefore well suited to study the physiological properties of horizontally extended retinal neurons as well as sensory signal transmission and integration across selected synapses.

We developed a horizontal slice preparation of the adult mammalian retina with the plane of sectioning parallel to the retinal surface. Dendritic fields of nonradially oriented retinal neurons were not truncated, allowing the study of signal processing with patch-clamp and imaging techniques.

電気生理学的研究のための成体マウスの網膜の水平スライスの調製

Vertical slice preparations are well established to study circuitry and signal transmission in the adult mammalian retina. The plane of sectioning in ...

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 ...