Name: an objective and reproducible test of olfactory learning and discrimination in mice
Uploaded: 2018-03-22T13:00:00
Description: Watch this Scientific Journal Video about An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice at JoVE.com

このプロトコルは、前作 (黄ら研究室内から適応8). ここで説明したすべてのメソッドは、動物のケアおよび使用委員会 (ACUC) の薬の Baylor の大学によって承認されています。によりマクネア医学研究所、NINDS 付与 R01NS078294 B.R.A.、IDDRC NIH グラント U54HD083092、jmp、NIDDK グラント F30DK112571 および組織プラスミノーゲンアクテベータ補助金 F31NS092435 CKM にサポートされます。

すべてのマウスは、NIH の基準に従ってベイラー大学の医学機関動物ケアおよび使用委員会によって承認されたプロトコルで使用されていた。マウスをこのプロトコルで使用されたすべての成体マウス (> 6 週齢) C57BL6/j の背景し、男性と女性の両方のマウスを含まれています。トレーニング/ステージング タスク後マウスが彼らのホームのケージに返されます。
1、建設やオ?...

<ol>
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B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+broadening+of+inhibitory+sensory+maps.">Developmental broadening of inhibitory sensory maps.</a> Nat Neurosci. 20 (2), 189 (2017).</li><li>Huang, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Task+learning+promotes+plasticity+of+interneuron+connectivity+maps+in+the+olfactory+bulb.">Task learning promotes plasticity of interneuron connectivity maps in the olfactory bulb.</a> J Neurosci. 36 (34), 8856-8871 (2016).</li><li>Arbuckle, E. P., Smith, G. D., Gomez, M. C., Lugo, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testing+for+odor+discrimination+and+habituation+in+mice.">Testing for odor discrimination and habituation in mice.</a> J Vis Sci. (99), e52615 (2015).</li><li>Zou, J., Wang, W., Pan, Y. W., Lu, S., Xia, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+to+measure+olfactory+behavior+in+mice.">Methods to measure olfactory behavior in mice.</a> Curr Protoc Toxicol. , 11-18 (2015).</li><li>Uchida, N., Takahashi, Y. K., Tanifuji, M., Mori, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Odor+maps+in+the+mammalian+olfactory+bulb:+domain+organization+and+odorant+structural+features.">Odor maps in the mammalian olfactory bulb: domain organization and odorant structural features.</a> Nat Neurosci. 3 (10), 1035 (2000).</li><li>Cang, J., Isaacson, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+whole-cell+recording+of+odor-evoked+synaptic+transmission+in+the+rat+olfactory+bulb.">In vivo whole-cell recording of odor-evoked synaptic transmission in the rat olfactory bulb.</a> J Neurosci. 23 (10), 4108-4116 (2003).</li><li>Parthasarathy, K., Bhalla, U. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laterality+and+symmetry+in+rat+olfactory+behavior+and+in+physiology+of+olfactory+input.">Laterality and symmetry in rat olfactory behavior and in physiology of olfactory input.</a> J Neurosci. 33 (13), 5750-5760 (2013).</li><li>Rajan, R., Clement, J. P., Bhalla, U. 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齧歯動物の嗅覚系は、感覚依存可塑性を勉強するユニークなモデルを提供します。報酬や罰臭気のペアに関連付けるためにネズミを訓練する嗅覚学習パラダイムをご紹介します。この学習を通じて下流の回路変更は (電気生理学、生体内神経イメージングなど) その後の実験で学ぶことができます。完了すると、マウスが 1 つ臭水報酬とタイムアウト罰別臭気に関連付ける単純な...

マウス OB、匂いの情報中枢神経系 (CNS) に初めて入る場所、経験依存性構造変化を研究するための優れたモデルを提供します。OB の迂遠は継続的に活動に依存した方法で大人生まれるニューロンを統合します。大人生まれるニューロン前駆細胞は側脳室1に隣接する脳室下帯をライン前駆細胞から切り分けます。OB に移行する時にこれらの神経前駆細胞生き残るため、差別化し抑制性顆粒細胞として統合またはアポトーシス2を受けます。細胞の運命の選択は、嗅覚学習3,4,5,6を含む嗅覚の活動によって影響されます。統合後学習によるシナプス変化は 2 週間重要に期間7,8中に顆粒細胞で発生します。したがって、嗅覚学習のための試金は、どのように経験依存的可塑性の影響構造と機能再編成熟脳回路を調べる際に役立ちます。
このプロトコルでは、オペラント条件づけを用いた嗅覚トレーニング方法の 1 つを提供しています。このタスクでは水を奪われたマウスは 1 つ臭 ("Go"臭気) 水報酬と別臭気 (「立入禁止」臭気) トライアル タイムアウト罰に関連付ける訓練されます。2-4 週間にわたって訓練段階の傾斜シリーズを通してマウスの進行状況。トレーニングが完了したら、マウスが離散、対応する動作 (移動試験水の報酬を求めておよび立入禁止の試験水の報酬を求めていない) と行ったり可否の臭気に対応 (図 1A)。トレーニングが完了した後、マウスをさらに化学的に類似の香りペアで挑戦できる差別をテストしたりに移行になる研究調査方法嗅覚学習改変の構造や OB の機能匂い弁別課題が報告されている、最も 2 つ匂い9,10間盗聴の数などの主観的な測定に依存してください。さらに、人間得点などのタスクのための必要性はまた時間がかかるです。このプロトコルで説明する行くか嗅覚学習作業では、公平な直接測定匂い弁別学習と嗅覚学習を提供しています。

<table border="0" cellpadding="0" cellspacing="0" style="width:1259px;" width="1258">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Glass vial</td>
			<td style="width:123px;">Qorpak</td>
			<td style="width:344px;">GLC-01016</td>
			<td style="width:576px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Silicon Tubing</td>
			<td>Thermo Scientific</td>
			<td>86000030</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">18 gauge needles</td>
			<td>BD</td>
			<td>305196</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">1-Butanol</td>
			<td>Sigma Aldrich</td>
			<td>437603</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Propionic Acid</td>
			<td>Sigma Aldrich</td>
			<td>402907</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Mouse Chamber</td>
			<td>Med Associates</td>
			<td>ENV-307W</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Chamber Floor</td>
			<td>Med Associates</td>
			<td>ENV-307W-GFW</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Water Port</td>
			<td>Med Associates</td>
			<td>ENV-313W</td>
			<td>Need two</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Odor stimulus</td>
			<td>Med Associates</td>
			<td>ENV-275</td>
			<td>Contain 2 valves to gate odor delivery&#160;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Odor Port</td>
			<td>Med Associates</td>
			<td>ENV-375W-NPP</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">USB Interface</td>
			<td>Med Associates</td>
			<td>DIG-703A-USB</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Desktop Computer with Windows 2000, XP, Vista, or 7</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Flow meter</td>
			<td>VWR</td>
			<td>97004-952</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Behavioral software</td>
			<td>Med Associates</td>
			<td>SOF-735</td>
			<td>This software, which runs each training stage, has now been replaced with Med-PC V</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Data Transfer software</td>
			<td>Med Associates</td>
			<td>SOF-731</td>
			<td>This software formats the data to Excel</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Training Software</td>
			<td>Med Associates</td>
			<td>DIG-703A-USB</td>
			<td>This software is used to program each training stage</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Water Valve</td>
			<td>Neptune Research</td>
			<td>225P012-11</td>
			<td>This valve is used to gate the water delivery. Need Two</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Odor Valve</td>
			<td>Neptune Research</td>
			<td>360P012-42</td>
			<td>This valve is used to gate the odor delivery. Need Two</td>
		</tr>
	</tbody>
</table>

an objective and reproducible test of olfactory learning and discrimination in mice

嗅覚はマウスで優勢な感覚モダリティと採餌、捕食動物の検出、交尾、子育てなど、多くの重要な動作に影響を与えます。重要なは、マウスを訓練して、小説臭の嗅覚回路機能への洞察力を提供するために具体的な行動応答を関連付けることができます。このプロトコルは、行くかどうかオペラント学習タスクにマウスのトレーニングの手順を詳しく説明します。このアプローチでマウスは 2-4 週間のために毎日自動試験の何百もの訓練、および新規行くかどうかにおいペアで嗅覚の差別を評価したり、構造や機能、嗅覚の匂い学習を変更する方法の研究に使用するテストできます。回路。さらに、マウスの嗅球 (OB) は大人生まれるニューロンの継続的な統合を備えています。興味深いことに、嗅覚学習増加生存とこれらの大人生まれるニューロンのシナプス接続します。したがって、このプロトコルは、神経細胞の生存と可塑性を仲介する学習と活動依存の要因を検討し他の生化学的、電気生理学的、およびイメージング技術と結合できます。

一度マウスの嗅覚学習タスクに学んだ、今報酬と罰小説臭ペアを関連付けることができます彼ら。これらの訓練を受けたマウスは通常約 50% で始まる進むかどうかのタスクでの精度。学習曲線の新しい臭気のペア (図 3A) として試作ブロックによって正しい割合をプロットできます。10 のブロック試験を実行する 30 分未満のほとんど?...

Watch this Scientific Journal Video about An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice at JoVE.com

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice

ここでは、匂い弁別学習をテストする連想学習タスクにマウスを養成します。このプロトコルでは、脳の学習誘起構造変化に関する研究も可能です。

嗅覚学習とマウスにおける差別の客観的かつ再現性のあるテスト

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and ...

The overall goal of this behavior protocol is to train mice so they associate one odor with a reward, and another odor with a punishment. This method can help answer key questions in the neuroscience field, such as learning, synaptic plasticity, and sensory discrimination. The main advantage of this technique is that once trained, mice perform hundreds of trials within hours for robust statistical sampling.The implications of this technique extend toward therapy or diagnosis of neurodegenerative diseases, because severity of olfactory dysfunction is predictive of subsequent cognitive decline in humans. Though this method can provide insight into olfaction, it can also be applied to other systems, such as vision or audition. Visual demonstration of this method is critical, as the initial steps for both setting up the behavior chamber and using the software are difficult to learn, because of different connecting ports and training steps.To begin, assemble a mouse chamber with chamber floor. Keep the training box in a low-traffic, dimly-lit area to avoid distractions. Then, drill each water port with a small hole on top, to allow an 18-gauge needle to dispense water inside the port.Fill a glass vial with odorant dissolved in mineral oil, and securely tighten the cap. Next, connect an 18-gauge needle to the silicone tubing. Pierce the glass vial cap, and connect the other end of the silicone tubing to the intake of the odor ports.After that, place each silicone tube into an odor valve. Connect the vacuum line to the odor ports. Attach two 10-milliliter syringes to a metal holding rod, and connect the tubing to the syringes.Subsequently, connect the other side of the tubing to an 18-gauge needle. Fit the needle into the drilled hole of the nose poke port on the mouse chamber, and connect the other end of the tubing to the water valve. Fill the two 10-milliliter syringes with rodent drinking water.Then connect an airflow meter to the air intake, and maintain airflow at three to five liters per minute. Connect the two water valves, two odor valves, two water ports, odor port, and power to the USB interface system. And lastly, connect the USB interface box to power.Next, adjust the vacuum suction to avoid cross-contamination of odors between trials, and use odor-specific tubing to connect odorant vials to the chamber. To prepare the mice for training, begin water restriction at 40 milliliters per kilogram per day. Weigh the mice daily to ensure they are above 80%of baseline weight, and keep all environmental factors constant throughout the protocol, including temperature, noise, and stray odors.Clean the cage before each mouse is transferred to it. Spray and wipe the chamber with 70%ethanol to minimize mouse odor distraction. For stage one training, configure the behavior box with a water port in the center, and with all side ports inaccessible.Place a mouse into the chamber and close it. Then, begin the stage one program. Consider this stage as complete when the mouse achieves 100 trials within 60 minutes.Remove the mouse from the chamber after 60 minutes, or 100 trials have been completed. For stage two training, configure this and every subsequent stage with two water ports on the sides and the odor port in the middle. Place a mouse into the chamber and close it.Then, begin the stage two program. Consider this stage as complete when a minimum of 40 trials are performed in 60 minutes, with at least 25%of water rewards received within five seconds of the center port nose poke. Remove the mouse once this stage has completed.For stage three training, connect the S-plus odor to the odor delivery controller. Place a mouse into the chamber and close it. Afterward, begin the stage three program.Consider this stage as complete when there are greater than 60 rewards within 60 minutes, and remove the mouse once the stage has competed. For stage four-A training, set the box configuration the same as previous stage three. Then, connect both S-plus and S-minus odor to the odor delivery controller.Place a mouse into the chamber and close it. Then, begin the stage four-A program. After the mouse completes 40 trials, switch the program to randomly deliver the odors.Remove the mouse once the stage has completed. For stage four-B training, make this stage identical to stage four-A. However, the time-out punishment for attempting a water reward after an S-minus odor is four seconds.Connect both S-plus and S-minus odor to the odor delivery controller. Place a mouse into the chamber. Close the mouse chamber and begin the stage four-B program.Consider this stage complete when there are more than 100 trials within 60 minutes, with accuracy more than 85%For go/no-go assay, set the box configuration the same as previous stage four-B. Connect both S-plus and S-minus odor to the odor delivery controller. Then, place a mouse into the chamber.Begin the stage go/no-go program and then expose the mouse that has learned the olfactory learning task to novel pairs of chemically similar odors. Once the mice have learned the olfactory learning task, they can associate novel odor pairs with reward and punishment. These trained mice normally begin with about 50%accuracy on the go/no-go task.The percentage correct can be plotted by trial block as a learning curve for novel odor pairs. Within 10 block trials, mice are able to correctly discriminate between odors, with greater than 85%accuracy. This shows that our protocol has successfully trained wild-type mice to associate one odor with a water reward and another with a time out punishment.Once the mice learn the task, odor pairs can be modified to increase or decrease the task difficulty. For example, decreasing the odor pair concentration increases the task difficulty. Mice are also able to remember previous learned odor pairs.After waiting seven days since learning the task, a recall test shows that wild-type mice can quickly remember previously-learned odor associations. While attempting this procedure, it's important to remember to ensure that the desired odorants are properly connected to the behavior box. It's especially important if multiple experimenters are using different odors.Following this procedure, other methods like odor mapping can be performed in order to answer additional questions like, How does learning influence the way sensory information is represented by the brain? After its development, this technique paved the way for researchers in the field of neuroscience to explore learning-induced neural changes in the mouse olfactory system. After watching this video, you should have a good understanding of how to teach mice to associate one odor with a reward and another odor with a punishment.Don't forget that working with sharp needles can be extremely hazardous, and precautions such as proper disposal of sharps should always be taken while performing this procedure.

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Low-stress Route Learning Using the Lashley III Maze in Mice

マウスでラシュリーIIIの迷路を用いた低ストレスルートの学習

Many behavior tests designed to assess learning and memory in rodents, particularly mice, rely on visual cues, food and/or water deprivation, or other ...

神経科学

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer&#39;s Disease Model Mice

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer's Disease Model Mice

アルツハイマー病モデルマウスにおける学習記憶障害のためのモリス水迷路試験

The Morris Water Maze (MWM) was first established by neuroscientist Richard G. Morris in 1981 in order to test hippocampal-dependent learning, including ...

T-maze Forced Alternation and Left-right Discrimination Tasks for Assessing Working and Reference Memory in Mice

マウスで作業していて、メモリの参照を評価するためのT-迷路強制交替と左右弁別課題

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in ...

Mapping Cortical Dynamics Using Simultaneous MEG/EEG and Anatomically-constrained Minimum-norm Estimates: an Auditory Attention Example

同時MEG / EEGおよび解剖学的に制約のある最小ノルム推定値を使用してマッピング皮質ダイナミクス：聴覚注意の例

Magneto- and electroencephalography (MEG/EEG) are neuroimaging techniques that provide a high temporal resolution particularly suitable to investigate the ...

Appetitive Associative Olfactory Learning in Drosophila Larvae

Appetitive Associative Olfactory Learning in Drosophila Larvae

で食欲連想嗅覚学習<em&gt;ショウジョウバエ</em&gt;幼虫

In the following we describe the methodological  details of appetitive associative olfactory learning in Drosophila larvae. The setup, in combination with ...

Drosophila Adult Olfactory Shock Learning

Drosophila Adult Olfactory Shock Learning

ショウジョウバエアダルト嗅覚ショックラーニング

Drosophila have been used in classical conditioning experiments for over 40 years, thus greatly facilitating our understanding of memory, including the ...

Ex Vivo Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb

Ex Vivo Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb

無傷の鋤鼻器官と副嗅球のex vivoでの準備

The mouse accessory olfactory system (AOS) is a specialized sensory pathway for detecting nonvolatile social odors, pheromones, and kairomones. The first ...

Whole Mount Labeling of Cilia in the Main Olfactory System of Mice

マウスの主嗅覚系における繊毛のホールマウントのラベリング

The mouse olfactory system comprises 6-10 million olfactory sensory neurons in the epithelium lining the nasal cavity. Olfactory neurons extend a single ...

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

の記録細胞内の電圧応答のための電気生理学的手法<em&gt;ショウジョウバエ</em&gt;光刺激に感光体と介在ニューロン<em&gt;インビボ</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

High Resolution Quantitative Synaptic Proteome Profiling of Mouse Brain Regions After Auditory Discrimination Learning

聴覚弁別学習した後、マウス脳領域の高分解能定量的シナプスプロテオームプロファイリング

The molecular synaptic mechanisms underlying auditory learning and memory remain largely unknown. Here, the workflow of a proteomic study on auditory ...