Name: imaging neuronal responses in slice preparations of vomeronasal organ expressing a genetically encoded calcium sensor
Uploaded: 2011-12-06T13:00:00
Description: Watch this Scientific Journal Video about Imaging Neuronal Responses in Slice Preparations of Vomeronasal Organ Expressing a Genetically Encoded Calcium Sensor at JoVE.com

We thank Andrea Moran together with members of Lab Animal Service Facility (LASF) at Stowers Institute for their excellent support on animal husbandry and technical services. This work is supported by funding from Stowers Institute and the NIH (NIDCD 008003) to CRY. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute on Deafness and Other Communication Disorders or the National Institutes of Health. U.S. patent pending for the tetO-G-CaMP2 mice for Stowers Institute, CRY and LM.

1。ソリューションの準備<ol><li>表によると10倍、R1、R2 10Xと10X R3ソリューションを準備します。 R1 <table border="1"><tbody><tr><td>化学物質</td><td> MW（g /モル） </td><td> mMの（1X） </td><td> 10X株式（G / L） </td></tr><tr><td> NaClを</td><td> 58.44 </td><td> 125 </td><td> 73.05 </td></tr><tr><td>塩化カリウム</td><td> 74.55 </td><td> 2.5 </td><td> 1.86 </td></tr><tr><td> MgCl 2の </td><td> 1Mストック</td><td> 1 </td><td> 10ミリリ?...

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鋤鼻受容体（VRS）の大半は、デュラックとアクセル5で、その発見以来、オーファン受容体として残る。これらの化学感覚受容体と動物の行動を媒介に自らの役割のためのフェロモンリガンドについてはよくわかっていない。今までは、リガンド/受容体の1ペアのみ、ESP1ペプチドおよびその同族体、Vmn2r116（V2Rp5）は、識別し、特定の社会的な情報19,23を伝えることが示されて?...

imaging neuronal responses in slice preparations of vomeronasal organ expressing a genetically encoded calcium sensor

鋤鼻器官（VNO）は、同じ種の1,2内の個人の、社会的な性と生殖に関するステータスに関する情報を運ぶ化学的感覚信号を検出します。これらの種内の信号を、フェロモンだけでなく、いくつかの捕食者3からの信号は、特異性と感度4の高レベルの鋤鼻感覚ニューロンを（VSNを）活性化する。 G -タンパク質共役型受容体、V1R、V2RとFPR 5月14日 、の少なくとも三つの異なるファミリーは化学的感覚手がかりの検出を仲介するVNOニューロンに発現されています。フェロモン情報はVNOによってエンコードされている方法を理解するために、それは様々な刺激に個々のVSNの応答プロファイルを分析し、これらの応答を仲介する特異的な受容体を識別することが重要です。 VNOのneuroepitheliaは、鋤骨のペアで囲まれています。 VNOの半盲管状の構造は、接続する1つの開放端を（鋤鼻ダクト）がある鼻腔。 VSNをはフェロモン手がかりが樹状ノブで発現する受容体と接触しているVNO、の内腔部分にその樹状突起を延長する。 VSNの細胞体は、頂端及び基底層にそれぞれ6-8を発現しV1RとV2Rを持つ擬似成層を形成する。いくつかの技術が感覚刺激4,12,15-19へのVSNの応答を監視するために利用されている。これらの技術の中で、急性スライス標本では、いくつかの利点を提供しています。最初に、解離VSNを3,17と比較して、スライス標本は、それらのネイティブ形態の神経細胞と細胞の樹状突起が比較的そのまま滞在を維持。第二に、VSNの細胞体は、全体の上皮およびホールマウント標本12,20に比べて電気生理学的試験とイメージング実験を可能にするVNOの冠状スライスに簡単にアクセスできます。第三に、このメソッドは、受容体の同定を可能にするために、分子クローニング技術と組み合わせることができます。感覚刺激が強いのCa 2受容体の活性化4,21の指標であるVSNをで+流入を誘発する。そこで我々は、トランスジェニックマウスを開発しているVSNを15,22を含む嗅覚ニューロンにおける発現するG - CaMP2。感度とプローブの遺伝的性質が大幅にCa 2 +動態のイメージング実験を促進する。このメソッドは、以前の研究4,21で使用される染料ロードプロセスを排除しています。我々はまた、VNOのスライスに様々な刺激の応用を可能にするリガンドの配信システムを採用しています。 2つの手法の組み合わせは、私たちは刺激の大きい数字に対応して同時に複数のニューロンを監視することができます。最後に、我々は画像処理を支援するための半自動解析パイプラインを確立している。

Watch this Scientific Journal Video about Imaging Neuronal Responses in Slice Preparations of Vomeronasal Organ Expressing a Genetically Encoded Calcium Sensor at JoVE.com

Imaging Neuronal Responses in Slice Preparations of Vomeronasal Organ Expressing a Genetically Encoded Calcium Sensor

The vomeronasal organ (VNO) detects intraspecies chemical signals that convey social and reproductive information. We have performed Ca2+ imaging experiments using transgenic mice expressing G-CaMP2 in VNO tissue. This approach allows us to analyze the complicated response patterns of the vomeronasal neurons to large numbers of pheromone stimuli.

遺伝的にコードされたカルシウムセンサーを発現する鋤鼻器のスライス標本のイメージング神経細胞応答

The vomeronasal organ (VNO) detects chemosensory signals that carry information about the social, sexual and reproductive status of the individuals within ...

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Neuroscience

Calcium Imaging of Odor-evoked Responses in the Drosophila Antennal Lobe

Calcium Imaging of Odor-evoked Responses in the Drosophila Antennal Lobe

の臭気誘発応答のカルシウムイメージングショウジョウバエ触角葉

The antennal lobe is the primary olfactory center in the insect brain and represents the anatomical and functional equivalent of the vertebrate olfactory ...

Imaging Pheromone Sensing in a Mouse Vomeronasal Acute Tissue Slice Preparation

マウス鋤鼻急性組織スライス標本のイメージングフェロモンセンシング

Peter Karlson and Martin L&uuml;scher used the term pheromone for the first time in 19591 to describe chemicals used for intra-species communication. ...

In vivo Neuronal Calcium Imaging in C. elegans

In vivo Neuronal Calcium Imaging in C. elegans

Cの生体内神経カルシウムイメージングでエレガンス

The nematode worm C. elegans is an ideal model organism  for relatively simple, low cost neuronal imaging in vivo. Its small transparent body and simple, ...

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 ...

Imaging Intracellular Ca2+ Signals in Striatal Astrocytes from Adult Mice Using Genetically-encoded Calcium Indicators

Imaging Intracellular Ca2+ Signals in Striatal Astrocytes from Adult Mice Using Genetically-encoded Calcium Indicators

イメージング細胞内Ca<sup&gt; 2+</sup遺伝的にエンコードされたカルシウム指標を用いて成体マウスからの線条体アスト​​ロサイト&gt;シグナル

Astrocytes display spontaneous intracellular Ca2+ concentration fluctuations ([Ca2+]i) and in several settings respond to neuronal excitation with ...

Recording Light-evoked Postsynaptic Responses in Neurons in Dark-adapted, Mouse Retinal Slice Preparations Using Patch Clamp Techniques

パッチクランプ技術を用いたダーク適応、マウス網膜スライス標本におけるニューロンにおける記録光誘発シナプス後応答

The retina is the gateway to the visual system. To understand visual signal processing mechanisms, we investigate retinal neural network functions. ...

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

遺伝的にコードされた蛍光電圧センサーの2種類のイメージング膜電位

Genetically encoded voltage indicators (GEVIs) have improved to the point where they are beginning to be useful for in vivo recordings. While the ultimate ...

Optical Control of a Neuronal Protein Using a Genetically Encoded Unnatural Amino Acid in Neurons

ニューロンに遺伝的にコードされた非天然アミノ酸を用いた神経タンパク質の光制御

Photostimulation is a noninvasive way to control biological events with excellent spatial and temporal resolution. New methods are desired to ...

Prolonged Incubation of Acute Neuronal Tissue for Electrophysiology and Calcium-imaging

電気生理学およびカルシウムイメージングのための急性神経組織の長期のインキュベーション

Acute neuronal tissue preparations, brain slices and retinal wholemount, can usually only be maintained for 6 - 8 h following dissection. This limits the ...

Two-photon Calcium Imaging in Neuronal Dendrites in Brain Slices

脳スライスにおける神経樹状突起における 2 光子カルシウム イメージング

Calcium (Ca2+) imaging is a powerful tool to investigate the spatiotemporal dynamics of intracellular Ca2+ signals in neuronal dendrites. Ca2+ ...