Name: morphological analysis of drosophila larval peripheral sensory neuron dendrites and axons using genetic mosaics
Uploaded: 2011-11-07T12:00:00
Description: Watch this Scientific Journal Video about Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics at JoVE.com

著者らは、資金調達のために理研に感謝。我々はまた、遺伝的および免疫組織化学プロトコルに関する議論のためにCagri Yalgin、キャロラインDelandre、とジェイパリッシュに感謝。

試薬の1.Preparation <ol><li>のCa + + -フリーHL3.1生理食塩水28を準備する。 <ol><li> mMで：70のNaCl、5のKCl、20のMgCl 2、10飽和NaHCO 3、5 HEPES、115スクロース、および5トレハロース、pHは7.2。滅菌濾過し、4℃で保存します。 注：CA + + - free溶液は、解剖時の筋収縮を防ぎます。 </li></ol></li><li>ポリ- L -リジン（PLL）カバースリップを行います。 <ol><li> -20℃で4.2ミ...

<ol>
	<li>Grueber, W. B., Jan, L. Y., Jan, Y. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tiling+of+the+Drosophila+epidermis+by+multidendritic+sensory+neurons.">Tiling of the Drosophila epidermis by multidendritic sensory neurons.</a> Development. 129, 2867-2878 (2002).</li><li>Crozatier, M., Vincent, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+multidendritic+neuron+differentiation+in+Drosophila:+the+role+of+Collier.">Control of multidendritic neuron differentiation in Drosophila: the role of Collier.</a> Dev Biol. 315, 232-242 (2008).</li><li>Hattori, Y., Sugimura, K., Uemura, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+expression+of+Knot/Collier,+a+transcriptional+regulator+of+the+EBF/Olf-1+family,+endows+the+Drosophila+sensory+system+with+neuronal+class-specific+elaborated+dendritic+patterns.">Selective expression of Knot/Collier, a transcriptional regulator of the EBF/Olf-1 family, endows the Drosophila sensory system with neuronal class-specific elaborated dendritic patterns.</a> Genes Cells. 12, 1011-1022 (2007).</li><li>Jinushi-Nakao, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Knot/Collier+and+cut+control+different+aspects+of+dendrite+cytoskeleton+and+synergize+to+define+final+arbor+shape.">Knot/Collier and cut control different aspects of dendrite cytoskeleton and synergize to define final arbor shape.</a> Neuron. 56, 963-978 (2007).</li><li>Sugimura, K., Satoh, D., Estes, P., Crews, S., Uemura, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+morphological+diversity+of+dendrites+in+Drosophila+by+the+BTB-zinc+finger+protein+abrupt.">Development of morphological diversity of dendrites in Drosophila by the BTB-zinc finger protein abrupt.</a> Neuron. 43, 809-822 (2004).</li><li>Li, W., Wang, F., Menut, L., Gao, F. 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ショウジョウバエ幼虫のDAニューロンモデルは、制御のニューロンの形態と回路形成そのメカニズムを調べる一つの優れた遺伝システムを提供します。 MARCMは、一般的に標識のためにと体DAニューロンのクローンを生成するために使用されます。 MARCMのために私達はどちらか汎神経（例えば、Gal4の C155）またはDAニューロン固有のドライバを使用してください。汎?...

<table border="1"><tr><th>試薬の名前</th><th>会社</th><th>カタログ番号</th><th>コメント（省略可能） </th></tr><tr><td> SZX16蛍光解剖顕微鏡（GFPHQフィルター付） </td><td>オリンポス</td><td> SZX16 </td><td></td></tr><tr><td>昆虫鉗子を生きる</td><td> FST </td><td> 26030〜10 </td><td></td></tr><tr><td> 26ミリメートルXの76ミリメートルうつ病のスライドガラス</td><td> Toshinriko（株） </td><td> T8 - R004 </td><td></td></tr><tr><td> Sylgard 184（またはSilpot 184） </td><td>ダウコーニング</td><td> 3097358-1004 </td><td></td></tr><tr><td>ポリ- L -リジン</td><td>シグマ</td><td> P - 1524 </td><td>この製品は、最も効果的であることが証明され</td></tr><tr><td> DPX封入剤</td><td>シグマ</td><td> 44581 </td><td></td></tr><tr><td>ウサギ抗GFP </td><td>インビトロジェン</td><td> - 11122 </td><td>希釈1:500 </td></tr><tr><td>ラット抗CD8 </td><td> Caltag </td><td> 5H10 </td><td>希釈1:200 </td></tr><tr><td>マウス抗CD2 </td><td> ABD serotec </td><td> MCA443R </td><td>希釈1:700 </td></tr><tr><td>マウス抗Fasciclin2 </td><td> DSHB </td><td> 1D4 </td><td>希釈午前1時10分</td></tr></table>

morphological analysis of drosophila larval peripheral sensory neuron dendrites and axons using genetic mosaics

神経系の発達には、正確なニューロンのクラス特有の樹状突起の発達と軸索の配線が続くニューロンの位置と同一の正確な仕様が必要です。最近ショウジョウバエの幼虫の末梢神経系（PNS）の樹状分枝（DA）の感覚ニューロンは、ニューロンの分化の両方の一般とクラス固有のメカニズムを解明するための強力な遺伝的モデルとなっている。 4つの主要なDAニューロンのクラス（I - IV）1があります。彼らは、増加する樹状突起の複雑さの順に名前が付けられ、それらの分化2-10の遺伝的制御のクラス固有の相違点を持っています。 1）それが2）、DAニューロン樹状突起が広がる、ショウジョウバエで利用できる強力な遺伝学的ツールを活用することができますので、DA感覚システムは、樹枝状形態の制御11から13の背後にある分子メカニズムを調べるために実用的なモデルです。光学的にCLEの下にだけ2次元のそれは容易にin vivoでの高分解能で可視化することAR幼虫のクチクラ、3）樹枝状形態におけるクラス固有の多様性は、単純な比較高度に分岐した樹枝状の木の形成を制御する重要な要素を見つけるために比較分析を容易に、そして4）樹状あずまや固定観念別のDAニューロンの形状は、形態学的統計分析を促進する。 DAニューロンの活動は、幼虫の運動中枢パターン発生器14から16の出力を変更します。別のDAニューロンのクラスは、異なる感覚モダリティを持っており、それらの活性化は、14,16-20異なる行動反応を誘発する。さらに別のクラスには、腹側神経索のショウジョウバエ幼虫の中枢神経系（VNC）21に型通りに軸索投射を送る。これらの予測は、樹状フィールド7,22の体壁のDAニューロンの感覚モダリティと位置の両方の地形表現で終了、23。 DA軸索突起のよって検査は、地形図7,22,23、だけでなく、簡単な回路を変調幼虫の運動14から17までの配線の基礎となるメカニズムを解明するために使用することができます。 ここではMARCM（抑制性細胞マーカーを持つモザイク解析）1,10,25とFLPアウト22,26,27技術（図1に要約）を介して24マーキングDAニューロンを生成し、遺伝的モザイクを分析する実用的なガイドを提示する。

Watch this Scientific Journal Video about Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics at JoVE.com

Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics

の樹状分枝感覚ニューロン<em&gt;ショウジョウバエ</em&gt;幼虫の末梢神経系は、一般とニューロンの分化のニューロンクラス特有のメカニズムの両方を解明するために有用なモデルです。我々は樹状突起分枝の神経細胞遺伝学的モザイクを生成して解析するための実用的なガイドを提示する。

の形態素解析ショウジョウバエ</em遺伝モザイクを使用>幼虫末梢感覚ニューロンの樹状突起と軸索

Nervous system development requires the correct specification of neuron position and identity, followed by accurate neuron class-specific dendritic ...

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Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics

In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons

In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons

ショウジョウバエ幼虫分節軸索内ではシナプス小胞の in vivo可視化で

Elucidating the mechanisms of axonal transport has shown to be very important in determining how defects in long distance transport affect different ...

Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall

Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall

免疫組織学的な感覚ニューロンの樹状突起における微小管の標識、気管、との筋肉ショウジョウバエ幼虫の体壁

To understand how differences in complex cell shapes are achieved, it is important to accurately follow microtubule organization. The Drosophila larval ...

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

使用して、マウス大脳皮質および海馬における可視化と樹状突起と棘の遺伝子操作<em&gt;子宮内で</em&gt;エレクトロポレーション

In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5. To date ...

Voltage-sensitive Dye Recording from Axons, Dendrites and Dendritic Spines of Individual Neurons in Brain Slices

軸索、樹状突起および脳スライスにおける個々のニューロンの樹状突起棘から電位感受性色素記録

Understanding the biophysical properties and functional organization of single neurons and how they process information is fundamental for understanding ...

Functional Analysis of the Larval Feeding Circuit in Drosophila

Functional Analysis of the Larval Feeding Circuit in Drosophila

幼虫給電回路の機能解析ショウジョウバエ</i

The serotonergic  feeding circuit in Drosophila  melanogaster larvae can be used to investigate neuronal substrates of  critical importance during the ...

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

における嗅覚受容ニューロンのホールマウント免疫標識<em&gt;ショウジョウバエ</em&gt;アンテナ

Odorant molecules bind to their target receptors in a precise and coordinated manner. Each receptor recognizes a specific signal and relays this ...

Production and Isolation of Axons from Sensory Neurons for Biochemical Analysis Using Porous Filters

多孔質フィルタを使用した生化学分析のための感覚ニューロンからの軸索の産生および単離

Neuronal axons use specific mechanisms to mediate extension, maintain integrity, and induce degeneration. An appropriate balance of these events is ...

Removal of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells

Removal of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells

の除去<em&gt;ショウジョウバエ</em感覚ニューロンおよび表皮細胞の免疫蛍光分析のための幼虫の切り身から&gt;筋肉組織

Drosophila larval dendritic arborization (da) neurons are a popular model for investigating mechanisms of neuronal morphogenesis. Da neurons develop in ...

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

勉強するリニアアガロースチャネルを使用します<em&gt;ショウジョウバエ</em&gt;幼虫クロールの動作

Drosophila larval crawling is emerging as a powerful model to study neural control of sensorimotor behavior. However, larval crawling behavior on flat ...

A Microbiomechanical System for Studying Varicosity Formation and Recovery in Central Neuron Axons

静脈瘤形成と中枢神経の軸索の回復を研究するための Microbiomechanical システム

Axonal varicosities are enlarged structures along the shafts of axons with a high degree of heterogeneity. They are present not only in brains with ...

の形態素解析ショウジョウバエ幼虫末梢感覚ニューロンの樹状突起と軸索