Name: measuring in vivo changes in extracellular neurotransmitters during naturally rewarding behaviors in female syrian hamsters
Uploaded: 2017-09-12T14:00:00.000+00:00
Duration: 10 min 23 s
Description: Watch this Scientific Journal Video about Measuring In Vivo Changes in Extracellular Neurotransmitters During Naturally Rewarding Behaviors in Female Syrian Hamsters at JoVE.com

著者は、Matlab コードを実行している彼のためのダニエルの Korus 学部と学部アレックス ベッチャー行動実験を実行している彼の援助に感謝したいと思います。このプロジェクトは、R.L.M. に NSF IOS 1256799 によって薬物乱用賞番号 T32DA007234 の下で健康の国民の協会の国立研究所でサポートされています。

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著者が明らかに何もありません。

比較的簡単ですが、この手法を採用する場合にいくつかの問題が発生します。まず、プローブの固定配置が正確でなければならない: マイクロダイアリシス プローブを取り巻く細胞外の環境のより広い半径でサンプルするとは異なりこの手法のみの直接接触に入って来る物質の計測が可能プローブ。第二に、繊維の幅が狭いため、炭素繊維の記録の場合破損が発生することができますと意図?...

目を覚まし行動中の動物の伝達物質の放出を測定する機能により、神経伝達の両方を生み出す機構と回路を調査するための強力なツールの時空間パターンの特定の動作をリンクする研究者自然リアルタイムでオペラント行動。歴史的に、脳1の細胞外の環境の両方の電気的反応性、非反応性の物質を測定するマイクロダイアリシスを採用されています。この技法は、先端が中空の半透膜2の小さいシャフトから成るマイクロダイアリシス プローブを細胞外液に類似のイオン組成の水溶液の連続的な流れを使用します。高速液体クロマトグラフィー (HPLC)、によってその後の分析のための間隔で収集される前に受動の拡散によって半透膜が出来る神経伝達物質または興味の他の検体プローブの挿入後、分析化学技術を区切るには、一般に利用を識別して異種混合物3のコンポーネントを定量化します。
マイクロダイアリシス興味の事実上すべての検体を測定する使用することができます敏感な手法ですが、時間の分解能は低く、およそ万分1,2分の最大サンプリング レート。高速スキャン サイクリックボルタンメトリー (FSCV)、電気活性種の酸化還元電位に依存する技術の発明は、細胞外液に興味の analyte の瞬時濃度近く解明できます。簡単に言うと (ロビンソンらを参照してください。4広範なレビューのため)、速い時間スケール4三角波ファッションに電圧を上下させる電極を適用します。電圧は、適切な範囲で、目的の化合物が繰り返し酸化して縮小しました。この酸化し削減微小交流電流を作成する電子の動きになります。スキャン レート酸化マイクロ秒単位で発生している化合物の還元とサブ秒スケールの場所を取る。現在の結果からのプローブによって作成された現在の背景を引く、現在プロットの各化合物に固有対の電圧を生成できます 1 つ。電圧振動のタイム スケールが知られているので、これらのデータは、現在のプロットを時間の関数として計算する使用できます。したがって、各酸化で転送された電子の数と還元反応が4を知られている限りは、化合物の相対濃度が決定されるかもしれない。
この化学的特異性と高い時間分解能 FSCV体内の化学成分の濃度変化を検出するための強力な技術を確認します。ただし、これらのマニホールドの利点にもかかわらず技術的なノウハウと高価な機器、セットアップにこのテクニックが必要。さらに、この手法を使用して、nonelectroactive 神経伝達物質 (例えばグルタミン酸) を測定できません。幸いなことに、これらの発明の商品化と同様、電気化学5の分野で技術の進歩は目を覚まし行動中の動物の非ゲルロボットの神経伝達物質を測定する比較的単純なアプローチを導入しています時間精度、テクニックを損なうことがなく酵素バイオ センサー技術として知られています。この手法は、あるゲルロボット過酸化水素応用可能性5 によって生成されたアンペロ メトリック酸化電流として検出される 2 枚の基板に関心の nonelectroactive 神経伝達物質の酵素的変換を使用してください。.市販バイオ センサー プローブ選択的に競争力のある内因性接しないの貢献を減らすことによって興味の検体を測定 (図 1参照)。グルタミン酸、場合共通 interferent アスコルビン酸 (AA) の貢献は競争力のある減測定電流に共同非ゲルロボット AA に変換するセンサーの活性酵素表面に AA オキシダーゼのローカライズdihydroascorbate と水。さらに、酵素層の下に存在負荷電ナフィオン膜は内因性アニオン性化合物を除外します。
この同じバイオ実験のセットアップは、FSCV のようにゲルロボットの神経伝達物質を測定できますが、代わりにそれは固定電位記録6を採用しています。FSCV で、振動電圧とは対照的で固定電位記録電圧は抑えている酸化還元で興味の analyte の潜在的です。このアプローチのターンキーの性質が化学物質の欠如を上回る圧倒的に 1 神経伝達物質に向かって傾斜する脳領域の同じ酸化還元電位、あるかもしれない複数の神経伝達物質として FSCV より少ない化学選択的ですが、特異性。
ほぼリアルタイムでゲルロボットと nonelectroactive の両方の神経伝達物質解放を測定し、特定の行動イベントにリンクする機能では、収束の伝達物質の放出を検討する機会を提供します。この原稿は、目を覚まし行動ハムスターにおける自然な報酬への応答でドーパミンやグルタミン酸の神経伝達を尋問するこのシステムの使用を詳しく説明します。本稿の目的は、性的行動その他の行動および実験パラダイムを調べるため実現の可能性を示すことを目標に、雌ハムスターの中にこの伝達物質の放出を測定するプロセスを詳しく説明することです。
ハムスターが電気化学的録音で使用するための理想的なモデル 歴史的に、ラットとマウスのモデルは、性行動の研究で採用されています。これらの齧歯動物種は耳がぴくぴくを追跡し、最終的にマウント女性7男性を誘惑するホッピング、ダーツなど多数の女性の勧誘行動を含む動的交尾シーケンスに従事します。(膣の浸透の有無にかかわらず) 男性による取付けは続きます、ほんの数秒、アクティブな勧誘行動を再開する前に、数秒間だけ中に女性を彼女の性的行動の姿勢で (前彎と呼ばれる) も行っています。この動作、不動の短い期間と散在している活動の高レベルの構成のパターンは動物の行動の神経伝達を測定する問題です。まず、神経活動とは無関係なアンペロ メトリックの録音で運動の成果物もあります。第二に、歩行が脳の特定領域の特定の神経伝達物質のリリースに関連付けられます。たとえば、ドーパミン放出は、背側と腹側線条体8、の9、マイクロダイアリシス ドーパミン精神刺激を次の測定のための基礎を形作った発見自発運動に結合されて管理10。Mos の女性-典型的な勧誘行動神経伝達の変化を総称して最後だけ性行動の明示的なコンポーネントに帰することは困難、t 齧歯動物の歩行活動の高レベルを含む、10 分性行動テストの大部分によって表されます分です。
女性の性行動の神経化学的プロファイルを分析するには、この演習は、性行動に伴う最小限の自発運動がある種を求めた。ハムスター (ん) の交尾のシーケンスは通常のラット及びマウス11見て勧誘行動の不足のため神経の録音に最適です。結果として、雌ハムスターが入力し、セッション12をテスト 10 分から 9 分の上向きの前弯姿勢を維持します。生体内で女性余分な歩行運動の欠如と男性と性的な相互作用のコンポーネントと関連付けることができる電気化学的録音が得られます。
ハムスター交尾発作 試験室に男性の刺激の動物の紹介の後、男性は最初彼女 (図 2A) をマウントする前に女性の性器調査 (AI) で従事します。マウントする男性に、女性は前弯で彼女は背中のアーチし、取付の男性は彼女の膣への陰茎のアクセスを得ることができますので、彼女の尾を偏向として知られている受容性姿勢を仮定しなければなりません。男性は両方の足 (図 2B) と彼女の後半部を握りしめる女性をマウントし、陰茎あたって (図 2C) を得るために試みに突っ込み始めます。男性は射精を最終的に達成する前に回数を intromit し同様 (挿入) することがなく女性がマウントされます。マウントと射精につながる intromissions のこのシーケンスは、「交尾試合」と呼ばれます。男性は、単一セッション内で複数交尾一続きがあります。

<table><tbody><tr><td>Nembutal</td><td>Oak Pharmaceuticals Inc.</td><td>76478-501-50</td><td>Pentobarbital sodium injection, USP. This lab uses 8.5 mg/100 g body weight, injected intraperotineally.&amp;nbsp;</td></tr><tr><td>Loxicom analgesic&amp;nbsp;</td><td>Norbrook Laboratories&amp;nbsp;</td><td>6451603670</td><td>NSAID antinflammatory and analgesic used for post-operative pain control. Generic: meloxicam.&amp;nbsp;</td></tr><tr><td>Enroflox antibiotic&amp;nbsp;</td><td>Norbrook Laboratories&amp;nbsp;</td><td>5552915411</td><td>Fluoroquinolone antibiotic for post-operative infection prevention. Generic: Enrofloxacin.</td></tr><tr><td>Beuthanasia-D&amp;nbsp;</td><td>Merck Animal Health</td><td>00061047305</td><td>Pentobarbital Sodium, Phenytoin Sodium euthanasia agent.</td></tr><tr><td>Bone screws</td><td>Pinnacle Technologies, Inc.</td><td>8111-16</td><td>1/8&quot; bone screw (Pkg. of 16) used to affix skull cap to skull.&amp;nbsp;</td></tr><tr><td>Dental acrylic (Bosworth Duz-All)</td><td>Bosworth&amp;nbsp;</td><td>166261C&amp;nbsp;</td><td>Self curing dental acrylic is used in construction of a skull cap to affix cannula and head mount to skull.</td></tr><tr><td>Hardware biosensor setup&amp;nbsp;</td><td>Pinnacle Technologies, Inc.</td><td>8400-K2</td><td>Pinnacle offers complete hardware kits for new users of our tethered biosensor system for rats. Kits include a commutator, preamplifier, and data conditioning and acquisition system</td></tr><tr><td>Base video computer package</td><td>Pinnacle Technologies, Inc.</td><td>9000-K1</td><td>The base computer package includes a preconfigured computer with ample hard disk storage, a high-definition monitor, a keyboard and mouse, an uninterruptible power supply, and all necessary cables.&amp;nbsp;</td></tr><tr><td>Video EQ700 EverFocus camera&amp;nbsp; package</td><td>Pinnacle Technologies, Inc.</td><td>&amp;nbsp;9000-K10&amp;nbsp;</td><td>EQ700 night vision capable box camera with independent IR source was obtained as part of Pinnacle video computer package. Dome camera (9000-K9) and HD camera (9000-K11) options are also available.&amp;nbsp;</td></tr><tr><td>Sirenia Acquisition software</td><td>Pinnacle Technologies, Inc.</td><td>Free--available to download from pinnaclet.com</td><td>Sirenia&amp;nbsp;Acquisition provides a single platform for recording data from any Pinnacle hardware system. The software features synchronization of all data streams, user-configurable settings, data consolidation, and multiple export options. In addition, the software includes basic review and analysis modules for biosensor recordings. Sirenia&amp;nbsp;delivers free ll-in-one software that is ideal for data acquisition and review.</td></tr><tr><td>Tethered rat&amp;nbsp;in vitro&amp;nbsp;calibration kit</td><td>Pinnacle Technologies, Inc.</td><td>7000-K2-T-BAS&amp;nbsp;</td><td>In order to relate the current changes measured by a biosensor to actual changes in analyte concentration, it is necessary to calibrate the biosensor prior to implantation into the animal. The process also confirms the integrity and selectivity of the sensors.&amp;nbsp;Calibration kit includes 20 mL jacketed beaker (#7058), 1/2&quot; by 1/8&quot; magnetic stir bar (#7059), right angle clamp (#7056), 2 prong single-adjustment clamp (#7055), 4-channel calibration preamplifer (#7053), and calibration holder (#7051).&amp;nbsp;</td></tr><tr><td>Stir plate&amp;nbsp;</td><td>Corning</td><td>6795-410D</td><td>Corning digital Stirrer, 5&quot; x 7&quot;, 120 VAC used to spin magnetic stirrer in jacketed beaker during &lt;em&gt;in vitro&lt;/em&gt; calibration of glutamate biosensors.</td></tr><tr><td>Water bath capable of closed loop circulation</td><td>PolyScience</td><td>8006A11B</td><td>PolyScience 8006A11B 6L Standard Digital Heated Circulating Bath, 120VAC water bath was used with plastic tubing to heat jacketed beaker to physiological temperature.&amp;nbsp;</td></tr><tr><td>Carbon fiber sensor with BASi rat cannulae</td><td>Pinnacle Technology, Inc.</td><td>7002-CFS</td><td>Carbon fiber electrode used for recording dopamine neurotransmission.</td></tr><tr><td>Ag/AgCl reference electrode</td><td>Pinnacle Technology, Inc.</td><td>7065</td><td>Necessary for carbon fiber recordings.</td></tr><tr><td>Glutamate biosensors&amp;nbsp;</td><td>Pinnacle Technology, Inc.</td><td>7001&amp;nbsp;</td><td>Enzymatic biosensor probe used for recording glutamatergic neurotransmission.</td></tr><tr><td>BASi guide cannulae</td><td>Pinnacle Technologies, Inc.</td><td>7030&amp;nbsp;</td><td>Guide cannulae implanted into brain region of interest to guide probe.</td></tr><tr><td>BASi cannula plastic headpiece for rats&amp;nbsp;</td><td>Pinnacle Technologies, Inc.</td><td>7011&amp;nbsp;</td><td>Headmount stabilizes probe and attaches to potentiostat.</td></tr></tbody></table>

measuring in vivo changes in extracellular neurotransmitters during naturally rewarding behaviors in female syrian hamsters

急速な時間スケールでの伝達物質の放出を測定する機能により、特定の動作または操作にリンクする神経伝達のパターン基になる機構と回路の解明で強力なツールです。マイクロダイアリシス法は、脳に興味のほぼすべての検体を測定する何十年も使用されています、この手法時間的解像度に限界があります。また、高速循環ボルタンメトリーは一時的に正確な非常に敏感です。ただし、技術的に困難な本法は興味の analyte の electroactivity に依存しているので、nonelectroactive 物質 (例えば、神経伝達物質グルタミン酸) を検出する可能性がなくなります。固定ポテンシャルをアンペロメトリーと時空間精度ゲルロボットと nonelectroactive の両方の神経伝達物質を測定する酵素バイオ センサーを組み合わせたターンキー システムの使用の詳細を説明します。これらの 2 つの強力な技術の組み合わせ、比較的簡単に強壮剤や一過性の神経伝達の測定により、複数の神経伝達物質の同時録音が可能します。本稿の目的は、ドーパミンやグルタミン酸神経伝達体内表示の究極の目標をハムスターの (すなわち、性行動) 当然のことながらやりがいのある動作を使用して測定のプロセスを示すため、その他の行動および実験パラダイムを調べるためこのアッセイの技術的可能性は。

電気化学および行動コーディング上記手法を使用して、この演習は、性行動の生体内で録音中にドーパミンやグルタミン酸の一過性変動・ トニックを特徴付けるし始めています。この方法論の一時的正確な方法によるより正確に性的行動中の神経伝達を特徴付けることができる我々同様、個々 の交尾行動 (例えばあたって、射精) の女性の受信中に対応?...

Watch this Scientific Journal Video about Measuring In Vivo Changes in Extracellular Neurotransmitters During Naturally Rewarding Behaviors in Female Syrian Hamsters at JoVE.com

Measuring In Vivo Changes in Extracellular Neurotransmitters During Naturally Rewarding Behaviors in Female Syrian Hamsters

自然やりがいのある行動の中にドーパミンやグルタミン酸高時間分解能のリリースを測定する炭素繊維電極と酵素バイオ センサー技術を使用して固定電位電流記録の使用の詳細を説明しますメスのハムスター。

当然やりがい雌ハムスターの行動中に細胞外の神経伝達物質の生体内変化を測定

The ability to measure neurotransmitter release on a rapid time scale allows patterns of neurotransmission to be linked to specific behaviors or ...

measuring-vivo-changes-extracellular-neurotransmitters-during

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Measuring In Vivo Changes in Extracellular Neurotransmitters During Naturally Rewarding Behaviors in Female Syrian Hamsters

Biopsychology

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10.1K ビュー.  University of Minnesota. 自然やりがいのある行動の中にドーパミンやグルタミン酸高時間分解能のリリースを測定する炭素繊維電極と酵素バイオ センサー技術を使用して固定電位電流記録の使用の詳細を説明しますメスのハムスター。

ビデオ: Measuring In Vivo Changes in Extracellular Neurotransmitters During Naturally Rewarding Behaviors in Female Syrian Hamsters

Comprehensive Analysis of Transcription Dynamics from Brain Samples Following Behavioral Experience

The encoding of experiences in the brain and the consolidation of long-term memories depend on gene transcription. Identifying the function of specific genes in encoding experience is one of the main objectives of molecular neuroscience. Furthermore, the functional association of defined genes with specific behaviors has implications for understanding the basis of neuropsychiatric disorders. Induction of robust transcription programs has been observed in the brains of mice following various behavioral manipulations. While some genetic elements are utilized recurrently following different behavioral manipulations and in different brain nuclei, transcriptional programs are overall unique to the inducing stimuli and the structure in which they are studied1,2.
In this publication, a protocol is described for robust and comprehensive transcriptional profiling from brain nuclei of mice in response to behavioral manipulation. The protocol is demonstrated in the context of analysis of gene expression dynamics in the nucleus accumbens following acute cocaine experience. Subsequent to a defined in vivo experience, the target neural tissue is dissected; followed by RNA purification, reverse transcription and utilization of microfluidic arrays for comprehensive qPCR analysis of multiple target genes. This protocol is geared towards comprehensive analysis (addressing 50-500 genes) of limiting quantities of starting material, such as small brain samples or even single cells.
The protocol is most advantageous for parallel analysis of multiple samples (e.g. single cells, dynamic analysis following pharmaceutical, viral or behavioral perturbations). However, the protocol could also serve for the characterization and quality assurance of samples prior to whole-genome studies by microarrays or RNAseq, as well as validation of data obtained from whole-genome studies.

This manuscript describes a protocol that applies comprehensive profiling for analysis of transcriptional programs induced in specific brain nuclei of rodents following behavioral paradigms. Herein, this approach is illustrated in the context of profiling genes induced in the nucleus accumbens (NAc) of mice following acute cocaine exposure, utilizing microfluidic qPCR arrays.

行動経験に続いて脳試料からの転写動態の網羅的解析

The encoding of experiences in the brain and the consolidation of long-term memories depend on gene transcription. Identifying the function of specific ...

行動学

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

変更された穴会は - マウスとラットにおける行動、認知と社会的相互作用を測定する

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

Quantifying Social Motivation in Mice Using Operant Conditioning

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered shuttle boxes were equipped with two operant levers (left and right) and a food receptacle in one chamber, which was then divided from the second chamber by an automated guillotine door covered by a wire grid. Different stimulus mice, rotated across testing days, served as a social stimulus behind the wire grid, and were only visible following the opening of the guillotine door. Test mice were trained to lever press in order to open the door and gain access to the stimulus partner for 15 sec. The number of lever presses required to obtain the social reward progressively increased on a fixed schedule of 3. Testing sessions ended after test mice stopped lever pressing for 5 consecutive minutes. The last reinforced ratio or breakpoint can be used as a quantitative measure of social motivation. For the second paradigm, test mice were trained to discriminate between left and right lever presses in order to obtain either a food reward or the social reward. Mice were rewarded for every 3 presses of each respective lever. The number of food and social rewards can be compared as a measurement of the value placed upon each reward. The ratio of each reward type can also be compared between mouse strains and the change in this ratio can be monitored within testing sessions to measure satiation with a given reward type. Both of these operant conditioning paradigms are highly useful for the quantification of social motivation in mouse models of autism and other disorders of social behavior.

This article describes a new protocol for the quantitative measurement of social motivation in mice using operant conditioning for a social reward. The protocol is useful for the measurement of social motivation in mouse models of autism and other disorders of social behavior.

オペラント条件付けを使用したマウスにおける社会的モチベーションを定量化します

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered ...

Survivable Stereotaxic Surgery in Rodents 

The ability to measure extracellular basal levels of neurotransmitters in the brain of awake animals allows for the determination of effects of different systemic challenges (pharmacological or physiological) to the CNS. For example, one can directly measure how the animal's midbrain dopamine projections respond to dopamine-releasing drugs like d-amphetamine or natural stimuli like food. In this video, we show you how to implant guide cannulas targeting specific sites in the rat brain, how to insert and implant a microdialysis probe and how to use high performance liquid chromatography coupled with electrochemical detection (HPLC-EC) to measure extracellular levels of oxidizable neurotransmitters and metabolites. Local precise introduction of drugs through the microdialysis probe allows for refined work on site specificity in a compound s mechanism of action. This technique has excellent anatomical and chemical resolution but only modest time resolution as microdialysis samples are usually processed every 20-30 minutes to ensure detectable neurotransmitter levels. Complementary ex vivo tools (i.e., slice and cell culture electrophysiology) can assist with monitoring real-time neurotransmission.

Survivable Stereotaxic Surgery in Rodents

The monitoring of extracellular neurotransmitter levels in distinct brain regions of freely moving animals offers insights on the link between neurotransmitter release and behavior. In vivo microdialysis coupled with electrochemical detection provides excellent anatomical and chemical resolution; and information on how basal neurotransmission is altered by pharmacological or physiological manipulations.

げっ歯類で存続可能な定位手術

The ability to measure extracellular basal levels of neurotransmitters in the brain of awake animals allows for the determination of effects of different ...

生物学

A Conflict Model of Reward-seeking Behavior in Male Rats

The present protocol describes a novel conflict task as a model of inhibitory control in rats. In this model, a natural rewarding stimulus (sexual stimulus) that represents a high-value reward, and the aversive stimuli (pins), are concurrently presented. The male rats have to climb or jump over the obstacle full of pins to approach and investigate the sexual partner. If the animal persists in their approaching behavior regardless of the aversive stimuli, it is considered as a maladaptive or risky reward-seeking behavior. The conflict task permits the evaluation of deficit in inhibitory control resulting from exposure to abused drug, such as morphine, or a stressful event. 
The main advantage of this model is that it provides a simple and quick way to discover the deficit in inhibitory control after exposure to opiate drugs or other stressful events. In addition to opiates, this behavioral model would also be useful for quickly discovering the inhibitory control deficits induced by other addictive drugs. However, the limitation is that the male rats' performance may be subject to exercising effects with repeated testing under this conflict task. In the future, one can hope that the individuals with the compulsive phenotype of reward-seeking behavior after exposure to opiates will be identified based on modifying this conflict model.

This conflict model is used to measure the impairment of inhibitory control after exposure to addictive drugs, or other factors that may influence inhibitory control. A sexual stimulus and an aversive obstacle are concurrently presented, thus male rats have to conquer the obstacle to approach the sexual reward.

雄ラットの報酬探索行動の競合モデル

The present protocol describes a novel conflict task as a model of inhibitory control in rats. In this model, a natural rewarding stimulus (sexual ...

Simultaneous Monitoring of Wireless Electrophysiology and Memory Behavioral Test as a Tool to Study Hippocampal Neurogenesis

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on animals and humans. Adult neurogenesis mechanism is also linked to memory and learning improvement. Traditionally, researchers used to assess learning and memory parameters in rodent models by behavioral tasks. Therefore, the simultaneous monitoring of behavioral changes and EEG is particularly interesting in correlating data between brain activity and task-related behaviors. However, most of the equipment required to perform both studies are either complex, expensive, or uses a wired setup network that hinders the natural animals&#8217; movement. In this study, EEG was recorded with a wireless electrophysiology device along with the execution of a novel object recognition task (NORT). The animal&#8217;s behavior was monitored simultaneously by a video tracking system. Both recordings were analyzed offline by their timestamps which were synched to link EEG signals with the animal&#8217;s actions. Subjects consist of adult Wistar rats after medium-term environmental enrichment treatment. Six skull screw electrodes were fixed in pairs on both hemispheres over frontal, central, and parietal regions and were referenced to an electrode located posterior of the nasal bone. NORT protocol consists of exposing the animal to two identical objects for 10 min. After 2 h and 24 h, one of the objects was replaced with a novel one. Exploration time for each object was monitored by a behavioral tracking software (BTS) and EEG data recording. The analysis of the EEG synced with behavioral data consists of estimations of alpha and beta relative band power and comparisons between novel object recognition versus familiar object exploration, between three experimental stages. In this manuscript, we have discussed electrodes manufacturing process, epidural electrodes implantation surgery, environmental enrichment protocol, NORT protocol, BTS setup, EEG &#8211; BTS coupling for simultaneous monitoring in real-time, and EEG data analysis based on automatic events detection.

The protocol presented here provides information on the simultaneous electroencephalography (EEG) and behavioral assessment in real-time. We have discussed all steps involved in this protocol as an attractive solution for researchers in many fields of neuroscience, particularly in learning and memory areas.

海馬神経新生研究ツールとしての無線電気生理学と記憶行動検査の同時モニタリング

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on ...

Environmental Modulations of the Number of Midbrain Dopamine Neurons in Adult Mice

Long-lasting changes in the brain or &#8216;brain plasticity&#8217; underlie adaptive behavior and brain repair following disease or injury. Furthermore, interactions with our environment can induce brain plasticity. Increasingly, research is trying to identify which environments stimulate brain plasticity beneficial for treating brain and behavioral disorders. Two environmental manipulations are described which increase or decrease the number of tyrosine hydroxylase immunopositive (TH+, the rate-limiting enzyme in dopamine (DA) synthesis) neurons in the adult mouse midbrain. The first comprises pairing male and female mice together continuously for 1 week, which increases midbrain TH+ neurons by approximately 12% in males, but decreases midbrain TH+ neurons by approximately 12% in females. The second comprises housing mice continuously for 2 weeks in &#8216;enriched environments&#8217; (EE) containing running wheels, toys, ropes, nesting material, etc., which increases midbrain TH+ neurons by approximately 14% in males. Additionally, a protocol is described for concurrently infusing drugs directly into the midbrain during these environmental manipulations to help identify mechanisms underlying environmentally-induced brain plasticity. For example, EE-induction of more midbrain TH+ neurons is abolished by concurrent blockade of synaptic input onto midbrain neurons. Together, these data indicate that information about the environment is relayed via synaptic input to midbrain neurons to switch on or off expression of &#8216;DA&#8217; genes. Thus, appropriate environmental stimulation, or drug targeting of the underlying mechanisms, might be helpful for treating brain and behavioral disorders associated with imbalances in midbrain DA (e.g. Parkinson&#8217;s disease, attention deficit and hyperactivity disorder, schizophrenia, and drug addiction).

This protocol describes two different environmental manipulations and a concurrent brain infusion protocol to study environmentally-induced brain changes underlying adaptive behavior and brain repair in adult mice.

成体マウスにおける中脳ドーパミンニューロンの数の環境モジュレーション

Long-lasting changes in the brain or &#8216;brain plasticity&#8217; underlie adaptive behavior and brain repair following disease or injury. Furthermore, ...

神経科学

Simultaneous Detection of c-Fos Activation from Mesolimbic and Mesocortical Dopamine Reward Sites Following Naive Sugar and Fat Ingestion in Rats

This study uses cellular c-fos activation to assess effects of novel ingestion of fat and sugar on brain dopamine (DA) pathways in rats. Intakes of sugars and fats are mediated by their innate attractions as well as learned preferences. Brain dopamine, especially meso-limbic and meso-cortical projections from the ventral tegmental area (VTA), has been implicated in both of these unlearned and learned responses. The concept of distributed brain networks, wherein several sites and transmitter/peptide systems interact, has been proposed to mediate palatable food intake, but there is limited evidence empirically demonstrating such actions. Thus, sugar intake elicits DA release and increases c-fos-like immunoreactivity (FLI) from individual VTA DA projection zones including the nucleus accumbens (NAC), amygdala (AMY) and medial prefrontal cortex (mPFC) as well as the dorsal striatum. Further, central administration of selective DA receptor antagonists into these sites differentially reduce acquisition and expression of conditioned flavor preferences elicited by sugars or fats. One approach by which to determine whether these sites interacted as a distributed brain network in response to sugar or fat intake would be to simultaneous evaluate whether the VTA and its major mesotelencephalic DA projection zones (prelimbic and infralimbic mPFC, core and shell of the NAc, basolateral and central-cortico-medial AMY) as well as the dorsal striatum would display coordinated and simultaneous FLI activation after oral, unconditioned intake of corn oil (3.5%), glucose (8%), fructose (8%) and saccharin (0.2%) solutions. This approach is a successful first step in identifying the feasibility of using cellular c-fos activation simultaneously across relevant brain sites to study reward-related learning in ingestion of palatable food in rodents.

The goal of this study is to identify reward-related distributed brain networks by delineating a reliable immunohistological technique using cellular c-fos activation to measure simultaneous changes in dopamine pathways and terminal sites after novel ingestion of fat and sugar in rats.

ラットにおけるナイーブ砂糖と脂肪の摂取後の中脳辺縁系と中間皮質ドーパミン報酬サイトからのc-Fosの活性化の同時検出

This study uses cellular c-fos activation to assess effects of novel ingestion of fat and sugar on brain dopamine (DA) pathways in rats. Intakes of sugars ...

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We demonstrate a simple way to tap into the fly reward system, modify experiences related to natural reward, and use voluntary ethanol consumption as a measure for changes in reward states. The approach serves as a relevant tool to study the neurons and genes that play a role in experience-mediated changes of internal state. The protocol is composed of two discrete parts: exposing the flies to rewarding and nonrewarding experiences, and assaying voluntary ethanol consumption as a measure of the motivation to obtain a drug reward. The two parts can be used independently to induce the modulation of experience as an initial step for further downstream assays or as an independent two-choice feeding assay, respectively. The protocol does not require a complicated setup and can therefore be applied in any laboratory with basic fly culture tools.

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for inducing rewarding and nonrewarding experiences in fruit flies (Drosophila melanogaster) using voluntary ethanol consumption as a measure for changes in reward states.

使用して報酬を求める行動の変化を測定するための簡単な方法<em&gt;キイロショウジョウバエ</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

In the central nervous system, the D1-alpha subtype receptor (Drd1&#945;) is the most abundant dopamine (DA) receptor, which plays a vital role in regulating neuronal growth and development. However, the mechanisms underlying Drd1&#945; receptor abnormalities mediating behavioral responses and modulating working memory function are still unclear. Using a novel RNA in situ hybridization assay, the current study identified dopamine Drd1&#945; receptor and tyrosine hydroxylase (TH) RNA expression from DA-related circuitry in the nucleus accumbens (NAc) area and substantia nigra region (SNR), respectively. Drd1&#945; expression in the NAc shows a &#34;discrete dot&#34; staining pattern. Clear sex differences in Drd1&#945; expression were observed. In contrast, TH shows a &#34;clustered&#34; staining pattern. Regarding TH expression, female rats displayed a higher signal expression per cell relative to male animals. The methods presented here provide a novel in situ hybridization technique for investigating changes in dopamine system dysfunction during the progression of central nervous system diseases.

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

Identification of dopamine D1-alpha receptor in the nucleus accumbens is critical for clarifying D1 receptor dysfunction during a central nervous system disease. We performed a novel RNA in situ hybridization assay to visualize single RNA molecules in a specific brain area.

当然やりがい雌ハムスターの行動中に細胞外の神経伝達物質の生体内変化を測定

要約

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