著者らは、デ·ラ·RECHERCHE専用サンテケベック（FRSQ）、カナダ衛生研究所（CIHR）、自然科学やカナダ工学研究会議（NSERC）フォンから給料賞、設備補助金、運転資金を承認したいと思いますコンコルディア大学研究チェアプログラム（CRUC）、ならびに博士ジェーン·スチュワートから本稿で思慮深いフィードバック。

1。動物ハウジング<ol><li> ケージ：個々のげっ歯類の走行輪活動を記録するために、各ケージは単齧歯類とrunning-ホイールを収納する必要があります。走行輪が濃縮の形と考えることができるので、任意の研究ではすべてのげっ歯類では、実行中のホイールに似てアクセスできる必要があります。 </li><li> 寝具の変更：彼らは動物との接触を最小限にするため、動物取扱と同様にケージや寝具の変化がすべての概日リズムを21から23に非透光効果を持つことができるので、メッシュ敷きのケージが理想的です。このようなトレイシステムの可用性にもかかわらず、寝具の変更は実験のクリティカルフェーズ中は避けるべきである。選択肢はもっとまれケージの変更を可能にするであろう長持ち寝具、または擬似ランダムなスケジュールで変更ケージを使用する方法があります。 </li><li> 隔離ボックス：ケージは光共、音減衰され隔離ボックスに保管してくださいntrolled、した風通しのよい。隔離ボックスのサイズと設定に応じて、各ボックス内のケージの数は、典型的には1から8の範囲であろう。 1つの絶縁ボックスに複数のケージを収容するとき、人は他の動物から来る様々な臭いや音が個々の動物の概日行動上の交絡効果を持つことができることを知っておく必要があります。これらの問題を回避するためには、分離ボックスあたり1ケージを収容するのを試みるべきです。 </li><li> 換気：十分に空気の流れは、げっ歯類のための快適な家庭環境ボックスを作るために不可欠です。各ボックス内のファンは内部に到達するボックス外からの光を防ぐために、フード付きでなければなりません。また、ファンは一般的に箱から空気を除去し、部屋に入って、それを爆破する。小さな光を通さない通気孔がいくつかのポイントから隔離ボックスを入力するように空気を可能にし、不快な風を防ぐのに役立ちます。十分な換気、隔離ボックス内の温度が（そこにあることを確認するために、）上のライトで、数時間のため閉鎖、それが収納されている部屋の温度と実質的に同一である必要があります。 </li><li> 照明：環境光の強さは、すべてのケージで同じでなければなりません。各ケージ上記と同様の場所にある単一の光をアレンジし、常に同じブランド/電球タイプのものをご使用ください。ケージレベルで適度な強度の照明（100から300ルクス）を使用します。むしろ概日システム、 それ自体 （ 例えばマスキング）よりも光に起因する行動に直接的な変化をもたらす可能性が高い高すぎる照明レベルを控えてください。 </li><li> 闇/ DIM赤い照明：それは暗闇の中で動物を（定数暗闇や夜間などで）処理するか、治療する必要がある場合には、暗視ゴーグルを使用する必要があります。概日システムは赤の波長に比較的鈍感であるため、あるいは、薄暗い赤い照明を使用することができます。あなたが使用する特定の赤色光はそれがアルテしないことを確実にするためにテストする必要がありますrは走行輪の活動（ 例えばマスキング）または（ 例えば、位相シフトを生成）体内時計を調整してください。 </li></ol> 2。データ収集（図1を参照してください - Vitalviewのハードウェア構成） <ol><li> 車輪を実行する：実行中の車輪の直径と人間工学は、使用24の量を変更します。したがって、マウス用の小型·軽量ホイールを使用し、ラットの大きい重いホイール。ホイールを洗浄してから再インストールするときは、車輪は遮るもののない、 &quot;ぐらつき&quot;はありませんし、録音マイクロスイッチは、ホイールの各ターンによって活性化されることをスピンすることができることを確認してください。 </li><li> マイクロスイッチ：実行中の車輪の各回転は、磁気的または機械的なマイクロスイッチを有効にする必要があります。マイクロスイッチから得た情報は、単一のチャネルを介して送信することができますし、時間をかけてビンのデータ（ 例えば、すべての2、5,6、または10分）コンピュータによって記録されます。 </li><li> コンピュータのハードウェア：当社runningの輪録音はVitalview、ミニマイテル（によって開発されたハードウェアとソフトウェアのプラットフォームで作られています<a href="http://www.minimitter.com/vitalview_software.cfm" target="_blank">http://www.minimitter.com/vitalview_software.cfm</a> ）。しかし、このようなActimetrics（ClockLabによって開発された、などの他のレコーディング·プラットホームがある<a href="http://www.actimetrics.com/ClockLab/" target="_blank">http://www.actimetrics.com/ClockLab/は</a> ）。両方のプラットフォームでは、単一のコンピュータのファイルに多くのシングル·チャンネル·ソースからのデータを（ 例えば実行中の1つの車輪によって活性化される単一のマイクロスイッチ）を一緒に持って来る。個々のチャネルからのデータは、その後、グラフ化され、後日、別々に分析することができます。 </li></ol> 3。データの録音<ol><li> ファイル：上記のソフトウェアプラットフォームは、個々のファイルは、実行中の各輪のレコードに対して作成されるように、単一のチャネルを分離するために使用することができます。このようなデータは特別にdesigneで最高の可視化とグラフ化されそのようなActiview（Minimitter、オレゴン州ベンド）、Circadia、またはすべてのピリオドとactogramsを生成することができますClocklab（Actimetrics、ウィルメット、イリノイ州）などのDプログラム。しかし、シングルチャネルのファイルも開いてこのようなエクセル（マイクロソフト社、レドモンド、ワシントン州）などの一般的な表計算プログラムを用いて分析することができる。 </li><li> 概日時間（CT）を計算する：CT 12は、定義によると、夜行性の齧歯類におけるランニングホイール活動の開始です。 24時間の日と並行して、慣例により、1日は24日周期の概日時間に分割されます。サーカディアンフリーランニング期間は壁時計によって測定されるよう24時間30分であればそれに応じて、CT 0 CT12後、約12時間15分を発生します。 </li></ol>

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特別な利害関係は宣言されません。

走行輪を使って毎日の活動リズムを監視することは夜行性の齧歯類におけるマスター概日時計の出力を評価するための最も一般的に使用され、信頼性の高い方法です。ホイールランニング活動、しかし、継続的に監視することができる行動や生理機能の多くの側面のひとつに過ぎません。実行中の輪活動の大半は夜間に行われますが、合計覚醒の30％以上が昼25,26中に発生します。他?...

<table border="1"&gt;<tbody&gt;<tr&gt;<td<strong試薬の&gt;名前</strong</td&gt;<td<strong&gt;会社概要</strong</td&gt;<td<strong&gt;カタログ番号</strong</td&gt;<td<strong&gt;コメント（オプション）</strong</td&gt;</tr&gt;<tr&gt;<td&gt; Vitalviewカード＆ソフトウェア</td&gt;<td&gt;ミニマイテル</td&gt;<td&gt;＃855-0030-00</td&gt;<td&gt;（ベンド、オレゴン州、米国）</td&gt;</tr&gt;<tr&gt;<td&gt; DP24データポート</td&gt;<td&gt;ミニマイテル</td&gt;<td&gt;＃840-0024-00</td&gt;<td&gt;（ベンド、オレゴン州、米国）</td&gt;</tr&gt;<tr&gt;<td&gt; QA4·モジュール</td&gt;<td&gt;ミニマイテル</td&gt;<td&gt;＃130-0050-00</td&gt;<td&gt;（ベンド、オレゴン州、米国）</td&gt;</tr&gt;<tr&gt;<td&gt;磁気スイッチ</td&gt;<td&gt;ミニマイテル</td&gt;<td&gt;＃130-0015-00</td&gt;<td&gt;（ベンド、オレゴン州、米国）</td&gt;</tr&gt;<tr&gt;<td&gt; C-50ケーブルアセンブリ</td&gt;<td&gt;ミニマイテル</td&gt;<td&gt;＃060-0045-10</td&gt;<td&gt;（ベンド、オレゴン州、米国）</td&gt;</tr&gt;<tr&gt;<tdホイールアセンブリを実行&gt;ラット</td&gt;<td&gt;ミニマイテル</td&gt;<td&gt;＃640-0700-00</td&gt;<td&gt;（ベンド、オレゴン州、米国）</td&gt;</tr&gt;<tr&gt;<td&gt;ケージとトレイ支持</td&gt;<td&gt;ミニマイテル</td&gt;<td&gt;＃640-0400-00</td&gt;<td&gt;（ベンド、オレゴン州、米国）</td&gt;</tr&gt;<tr&gt;<td&gt;使用可能にケージを切り取る</td&gt;<td&gt;ミニマイテル</td&gt;<td&gt;＃664-2154-00</td&gt;<td&gt;（ベンド、オレゴン州、米国）</td&gt;</tr&gt;<tr&gt;<tdケージ用&gt;グリッド床</td&gt;<td&gt;ミニマイテル</td&gt;<td&gt;＃676-2154-00</td&gt;<td&gt;（ベンド、オレゴン州、米国）</td&gt;</tr&gt;<tr&gt;<td&gt;廃棄トレイ</td&gt;<td&gt;ミニマイテル</td&gt;<td&gt;＃684-2154-00</td&gt;<td&gt;（ベンド、オレゴン州、米国）</td&gt;</tr&gt;<tr&gt;<td&gt;ランプハウジング</td&gt;<td&gt;マイクロライト·サイエンティフィック</td&gt;<td&gt;＃R-101</td&gt;<td&gt;（トロント、ON、カナダ）</td&gt;</tr&gt;<tr&gt;<td&gt; 4W蛍光灯</td&gt;<td&gt;マイクロライト·サイエンティフィック</td&gt;<td&gt;＃F4T5/CW</td&gt;<td&gt;（トロント、ON、カナダ）</td&gt;</tr&gt;<tr&gt;<td&gt;アイソレーションチャンバー</td&gt;<td&gt;カスタム構築された</td&gt;<td</td&gt;<td&gt; 28 &quot;H×20&quot;幅x 28 &quot;D½&quot;ブラックメラミン。</td&gt;</tr&gt;</tbody&gt;</table

recording and analysis of circadian rhythms in running-wheel activity in rodents

げっ歯類は、ホームケージで実行されているホイールへの無料アクセスを提供していたとき、このホイールの自主的な使用は、一日1-5時間に依存します。ラット、ハムスター、マウスなどの夜行性の齧歯類では、夜の間にアクティブであり、日中は比較的不活性。他の多くの行動や生理的な措置は、生活リズムを示すが、げっ歯類では、実行中の輪活動は視床下部の視交叉上核（SCN）、マスター概日時計の出力は特に信頼性と便利な尺度として役立つ。一般的には、エントレインメントと呼ばれるプロセスを介して、実行中の輪活動の毎日のパターンが自然環境の明暗サイクル（;：12時間-暗例えば 12時間-光LDサイクル）に合わせます。しかし概日リズムは内因性に一定の暗闇の中で〜24時間の期間を示し、持続する行動のパターンを生成されます。このように、LDのサイクルが存在しない場合には、走行輪活動の記録と分析ができます主観的な時間帯を決定するために使用される。これらのリズムが体内時計によって指示されているので、主観的な時刻は概日時間（CT）と呼ばれています。これとは対照的に、LDのサイクルが存在しているとき、環境のLDサイクルによって決定される時刻は、zeitgeberタイム（ZT）と呼ばれています。 走行輪活動の概日リズムは、典型的には、SCNクロック6から8にリンクされているが、脳と身体9-14の他の多くの地域における概日振動子も毎日の活動リズムの調節に関与している可能性があります。例えば、食品先行アクティビティの生活リズムは、SCN 15,16を必要とせず、その代わりに、余分-SCN発振器17から20の活性の変化と相関している。したがって、実行中の輪活動の記録は、マスターSCNクロックの出力についてだけでなく、余分-SCN発振器の活動のみならず重要な行動情報を提供することができる。我々descrを下機器や実験用げっ歯類の概日歩行活動リズムを記録し、分析して表示するために使用されるメソッドをIBE。

<ol><li> コンピュータプログラム：専用のコンピュータプログラムは、一般的にactogramsと概日周期の計算の生成に使用されています。これらのプログラムが含まれますが、（Minimitter、オレゴン州ベンド）とCircadia Actiview、これらに限定されない。 </li><li> Actograms：Actogramsは、running-ホイール活動の毎日のパターンの写実的な実例を提供します。シングルプロット（x軸= 24時間）とダブルプ...

Watch this Scientific Journal Video about Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents at JoVE.com

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

哺乳類における自主的なホイールランニング活動の概日リズムはしっかりと脳内のマスタ·クロックの分子振動に結合されている。このように、この動作の生活リズムは、この体内時計の機能に関する遺伝的、薬理学的、および環境要因の影響を研究するために使用することができます。

げっ歯類におけるランニングホイール活動の概日リズムの記録および分析

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

recording-analysis-circadian-rhythms-running-wheel-activity

Research

JoVE Journal

Neuroscience

21.1K ビュー.  McGill University. 哺乳類における自主的なホイールランニング活動の概日リズムはしっかりと脳内のマスタ·クロックの分子振動に結合されている。このように、この動作の生活リズムは、この体内時計の機能に関する遺伝的、薬理学的、および環境要因の影響を研究するために使用することができます。

ビデオ: Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) pacemakers or clocks. Malfunctions in the human circadian system are associated with numerous diseases or disorders. Much progress towards our understanding of the mechanisms underlying circadian rhythms has emerged from genetic screens whereby an easily measured behavioral rhythm is used as a read-out of clock function. Studies using Drosophila have made seminal contributions to our understanding of the cellular and biochemical bases underlying circadian rhythms. The standard circadian behavioral read-out measured in Drosophila is locomotor activity. In general, the monitoring system involves specially designed devices that can measure the locomotor movement of Drosophila. These devices are housed in environmentally controlled incubators located in a darkroom and are based on using the interruption of a beam of infrared light to record the locomotor activity of individual flies contained inside small tubes. When measured over many days, Drosophila exhibit daily cycles of activity and inactivity, a behavioral rhythm that is governed by the animal's endogenous circadian system. The overall procedure has been simplified with the advent of commercially available locomotor activity monitoring devices and the development of software programs for data analysis. We use the system from Trikinetics Inc., which is the procedure described here and is currently the most popular system used worldwide. More recently, the same monitoring devices have been used to study sleep behavior in Drosophila. Because the daily wake-sleep cycles of many flies can be measured simultaneously and only 1 to 2 weeks worth of continuous locomotor activity data is usually sufficient, this system is ideal for large-scale screens to identify Drosophila manifesting altered circadian or sleep properties.

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

We describe procedures for recording daily locomotor activity rhythms of Drosophila and subsequent data analysis. Locomotor activity rhythms are a reliable behavioral output of animal circadian clocks and are used as the standard readout of clock function when studying circadian mutants or examining how the environment regulates the circadian system.

に概日リズムと睡眠のパラメータを研究するために運動活性を測定するショウジョウバエ</em

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) ...

神経科学

Measuring Circadian and Acute Light Responses in Mice using Wheel Running Activity

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2. They are responsible for timing our sleep/wake cycles and hormone secretion. Since this timing is not precisely 24-hours, it is synchronized to the solar day by light input. This is accomplished via photic input from the retina to the suprachiasmatic nucleus (SCN) which serves as the master pacemaker synchronizing peripheral clocks in other regions of the brain and peripheral tissues to the environmental light dark cycle3-7. The alignment of rhythms to this environmental light dark cycle organizes particular physiological events to the correct temporal niche, which is crucial for survival8. For example, mice sleep during the day and are active at night. This ability to consolidate activity to either the light or dark portion of the day is referred to as circadian photoentrainment and requires light input to the circadian clock9. Activity of mice at night is robust particularly in the presence of a running wheel. Measuring this behavior is a minimally invasive method that can be used to evaluate the functionality of the circadian system as well as light input to this system. Methods that will covered here are used to examine the circadian clock, light input to this system, as well as the direct influence of light on wheel running behavior.

This article will review methods that can be used to determine circadian function and light responsiveness in mice.

ホイール実行中のアクティビティを使用して、マウスの概日リズムと急性ライト応答を測定する

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2.  They ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects1-11. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design.
Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (<a href="http://www.neuronexustech.com/" >http://www.neuronexustech.com/</a>; <a href="http://www.sbmicrosystems.com/">http://www.sbmicrosystems.com/</a>; <a href="http://www.acreo.se/">http://www.acreo.se/</a>) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.

We describe methods for large-scale recording of multiple single units and local field potential in behaving rodents with silicon probes. Drive fabrication, probe attachment to the drive and probe implantation processes are illustrated in sufficient details for easy replication.

動作げっ歯類の可動シリコンプローブによるニューロンの大規模なレコーディング

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

A Fully Automated and Highly Versatile System for Testing Multi-cognitive Functions and Recording Neuronal Activities in Rodents

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be investigated in a single task sequence. The unique feature of this system is a custom-made, acoustically transparent chamber that eliminates many of the issues associated with auditory cue control in most commercially available chambers. The ease with which operant devices can be added or replaced makes this system quite versatile, allowing for the implementation of a variety of auditory, visual, and olfactory behavioral tasks. Automation of the system allows fine temporal (10 ms) control and precise time-stamping of each event in a predesigned behavioral sequence. When combined with a multi-channel electrophysiology recording system, multiple cognitive brain functions, such as motivation, attention, decision-making, patience, and rewards, can be examined sequentially or independently.

In this report, we present a fully automated and highly versatile system capable of simultaneously testing multi-cognitive behaviors and recording neuronal activities for rodents.

マルチ認知機能のテストとげっ歯類の神経活動を記録するための完全に自動化された汎用性の高いシステム

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be ...

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from individual neurons are critical for understanding how neural circuits function under natural conditions. Traditionally, these recordings have been performed 'blind', meaning the identity of the recorded cell is unknown at the start of the recording. Cellular identity can be subsequently determined via intracellular1, juxtacellular2 or loose-patch3 iontophoresis of dye, but these recordings cannot be pre-targeted to specific neurons in regions with functionally heterogeneous cell types. Fluorescent proteins can be expressed in a cell-type specific manner permitting visually-guided single-cell electrophysiology4-6. However, there are many model systems for which these genetic tools are not available. Even in genetically accessible model systems, the desired promoter may be unknown or genetically homogenous neurons may have varying projection patterns. Similarly, viral vectors have been used to label specific subgroups of projection neurons7, but use of this method is limited by toxicity and lack of trans-synaptic specificity. Thus, additional techniques that offer specific pre-visualization to record from identified single neurons in vivo are needed. Pre-visualization of the target neuron is particularly useful for challenging recording conditions, for which classical single-cell recordings are often prohibitively difficult8-11. The novel technique described in this paper uses retrograde transport of a fluorescent dye applied using tungsten needles to rapidly and selectively label a specific subset of cells within a particular brain region based on their unique axonal projections, thereby providing a visual cue to obtain targeted electrophysiological recordings from identified neurons in an intact circuit within a vertebrate CNS.
The most significant novel advancement of our method is the use of fluorescent labeling to target specific cell types in a non-genetically accessible model system. Weakly electric fish are an excellent model system for studying neural circuits in awake, behaving animals12. We utilized this technique to study sensory processing by "small cells" in the anterior exterolateral nucleus (ELa) of weakly electric mormyrid fish. "Small cells" are hypothesized to be time comparator neurons important for detecting submillisecond differences in the arrival times of presynaptic spikes13. However, anatomical features such as dense myelin, engulfing synapses, and small cell bodies have made it extremely difficult to record from these cells using traditional methods11, 14. Here we demonstrate that our novel method selectively labels these cells in 28% of preparations, allowing for reliable, robust recordings and characterization of responses to electrosensory stimulation.

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

逆行性蛍光標識は、特定された神経細胞から標的細胞外シングルユニット記録を可能にします<em生体内で&gt;</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

ローカスの神経活動を特徴づけるためにマルチユニット記録方法（<em&gt; Schistocercaアメリカーナ</em&gt;）嗅覚回路

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for using blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) to image rodents with simultaneous DBS. DBS fMRI presents a number of technical challenges, including accuracy of electrode implantation, MR artifacts created by the electrode, choice of anesthesia and paralytic to minimize any neuronal effects while simultaneously eliminating animal motion, and maintenance of physiological parameters, deviation from which can confound the BOLD signal.&#160;Our laboratory has developed a set of procedures that are capable of overcoming most of these possible issues. For electrical stimulation, a homemade tungsten bipolar microelectrode is used, inserted stereotactically at the stimulation site in the anesthetized subject. In preparation for imaging, rodents are fixed on a plastic headpiece and transferred to the magnet bore. For sedation and paralysis during scanning, a cocktail of dexmedetomidine and pancuronium is continuously infused, along with a minimal dose of isoflurane; this preparation minimizes the BOLD ceiling effect of volatile anesthetics. In this example experiment, stimulation of the subthalamic nucleus (STN) produces BOLD responses which are observed primarily in ipsilateral cortical regions, centered in motor cortex. Simultaneous DBS and fMRI allows the unambiguous modulation of neural circuits dependent on stimulation location and stimulation parameters, and permits observation of neuronal modulations free of regional bias. This technique may be used to explore the downstream effects of modulating neural circuitry at nearly any brain region, with implications for both experimental and clinical DBS.

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

げっ歯類での同時のfMRIと脳深部刺激

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments

Afferent nerves not only convey information concerning normal physiology, but also signal disturbed homeostasis and pathophysiological processes of the different organ systems from the periphery towards the central nervous system. As such, the increased activity or &#39;sensitization&#39; of mesenteric afferent nerves has been allocated an important role in the pathophysiology of visceral hypersensitivity and abdominal pain syndromes.
Mesenteric afferent nerve activity can be measured in vitro in an isolated intestinal segment that is mounted in a purpose-built organ bath and from which the splanchnic nerve is isolated, allowing researchers to directly assess nerve activity adjacent to the gastrointestinal segment. Activity can be recorded at baseline in standardized conditions, during distension of the segment or following the addition of pharmacological compounds delivered intraluminally or serosally. This technique allows the researcher to easily study the effect of drugs targeting the peripheral nervous system in control specimens; besides, it provides crucial information on how neuronal activity is altered during disease. It should be noted however that measuring afferent neuronal firing activity only constitutes one relay station in the complex neuronal signaling cascade, and researchers should bear in mind not to overlook neuronal activity at other levels (e.g., dorsal root ganglia, spinal cord or central nervous system) in order to fully elucidate the complex neuronal physiology in health and disease.
Commonly used applications include the study of neuronal activity in response to the administration of lipopolysaccharide, and the study of afferent nerve activity in animal models of irritable bowel syndrome. In a more translational approach, the isolated mouse intestinal segment can be exposed to colonic supernatants from IBS patients. Furthermore, a modification of this technique has been recently shown to be applicable in human colonic specimens.

In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments

Mesenteric afferent nerves convey information from the gastrointestinal tract towards the brain regarding normal homeostasis as well as pathophysiology. Gastrointestinal afferent nerve activity can be assessed by mounting isolated intestinal segments with attached afferent nerves into an organ bath, isolating the nerve, and assessing basal as well as stimulated activity.

マウス空腸および結腸セグメントにおける腸間膜求心性神経活動のin vitro記録で

Afferent nerves not only convey information concerning normal physiology, but also signal disturbed homeostasis and pathophysiological processes of the ...

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic tectum, is a popular model to study how neural circuits self-assemble. The ability to carry out whole cell patch clamp recordings from tectal neurons and to record RGC-evoked responses, either in vivo or using a whole brain preparation, has generated a large body of high-resolution data about the mechanisms underlying normal, and abnormal, circuit formation and function. Here we describe how to perform the in vivo preparation, the original whole brain preparation, and a more recently developed horizontal brain slice preparation for obtaining whole cell patch clamp recordings from tectal neurons. Each preparation has unique experimental advantages. The in vivo preparation enables the recording of the direct response of tectal neurons to visual stimuli projected onto the eye. The whole brain preparation allows for the RGC axons to be activated in a highly controlled manner, and the horizontal brain slice preparation allows recording from across all layers of the tectum.

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

In this paper, we discuss three brain preparations used for whole cell patch clamp recording to study the retinotectal circuit of Xenopus laevis tadpoles. Each preparation, with its own specific advantages, contributes to the experimental tractability of the Xenopus tadpole as a model to study neural circuit function.

準備やアフリカツメガエルの視蓋ニューロンの全体セル パッチ ・ クランプ記録のためのプロトコル

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in neurocognitive disorders. Despite the popularization of prepulse inhibition (PPI) in recent years, many current protocols promote using a percent of control measure, thereby precluding the assessment of temporal processing. The present study used cross-modal PPI and gap prepulse inhibition (gap-PPI) to demonstrate the benefits of employing a range of interstimulus intervals (ISIs) to delineate effects of sensory modality, psychostimulant exposure, and age. Assessment of sensory modality, psychostimulant exposure, and age reveals the utility of an approach varying the interstimulus interval (ISI) to establish the shape of the ISI function, including increases (sharper curve inflections) or decreases (flattening of the response amplitude curve) in startle amplitude. Additionally, shifts in peak response inhibition, suggestive of a differential sensitivity to the manipulation of ISI, are often revealed. Thus, the systematic manipulation of ISI affords a critical opportunity to evaluate temporal processing, which may reveal the underlying neural mechanisms involved in neurocognitive disorders.

Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.