昆虫の解剖学の知識を共有するためヨッヘン ・ ツァイル、ポール ・ クーパーとビルギット グライナーに感謝しております、テクニックのいくつかを示すため、我々 はここで記述しました。オーストラリア国立大学で高度な顕微鏡と顕微鏡ユニット MQU でセンターで支持と有能なスタッフに感謝しております。この作品は、オーストラリアの研究評議会 (DE120100019、FT140100221、DP150101172) から周波数と補助金に大学院奨学金によって支えられました。

<ol><li>Zeil, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Visual+homing%3A+an+insect+perspective%5BTitle%5D)+AND+%22Curr.+Opin.+Neurobiol%22%5BJournal%5D)">Visual homing: an insect perspective.</a> Curr. Opin. Neurobiol. 22, 285-293 (2012).</li>
<li>Wehner, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Desert+ant+navigation%3A+how+miniature+brains+solve+complex+tasks%5BTitle%5D)+AND+%22J.+Comp.+Physiol.+A%22%5BJournal%5D)">Desert ant navigation: how miniature brains solve complex tasks.</a> J. Comp. Physiol. A. 189, 579-588 (2003).</li>
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<li>Bulova, S., Purce, K., Khodak, P., Sulger, E., O'Donnell, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Into+the+black+and+back%3A+the+ecology+of+brain+investment+in+Neotropical+army+ants+%28Formicidae%3A+Dorylinae%29%5BTitle%5D)+AND+%22Naturwissen%22%5BJournal%5D)">Into the black and back: the ecology of brain investment in Neotropical army ants (Formicidae: Dorylinae).</a> Naturwissen. 103, 3-4 (2016).</li>
<li>Narendra, A., Reid, S. F., Hemmi, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+twilight+zone%3A+ambient+light+levels+trigger+activity+in+primitive+ants%5BTitle%5D)+AND+%22Proc.+R.+Soc.+B%22%5BJournal%5D)">The twilight zone: ambient light levels trigger activity in primitive ants.</a> Proc. R. Soc. B. 277, 1531-1538 (2010).</li>
<li>Narendra, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Caste-specific+visual+adaptations+to+distinct+daily+activity+schedules+in+Australian+Myrmecia+ants%5BTitle%5D)+AND+%22Proc.+R.+Soc.+B%22%5BJournal%5D)">Caste-specific visual adaptations to distinct daily activity schedules in Australian Myrmecia ants.</a> Proc. R. Soc. B. 278, 1141-1149 (2011).</li>
<li>Moser, J., et al. <a target="_blank" href="https://www.treesearch.fs.fed.us/pubs/9853">Eye size and behaviour of day-and night-flying leafcutting ant alates.</a> J. Zool. 264, 69-75 (2004).</li>
<li>Stöckl, A. L., Ribi, W. A., Warrant, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adaptations+for+nocturnal+and+diurnal+vision+in+the+hawkmoth+lamina%5BTitle%5D)+AND+%22J.+Comp.+Neurol%22%5BJournal%5D)">Adaptations for nocturnal and diurnal vision in the hawkmoth lamina.</a> J. Comp. Neurol. 524, 160-175 (2016).</li>
<li>Zeil, J. <a target="_blank" href="https://link.springer.com/article/10.1007/BF00605027">Sexual dimorphism in the visual system of flies: the compound eyes and neural superposition in Bibionidae (Diptera).</a> J. Comp. Physiol. A. 150, 379-393 (1983).</li>
<li>Dacke, M., Nordström, P., Scholtz, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Twilight+orientation+to+polarised+light+in+the+crepuscular+dung+beetle+Scarabaeus+zambesianus%5BTitle%5D)+AND+%22J.+Exp.+Biol%22%5BJournal%5D)">Twilight orientation to polarised light in the crepuscular dung beetle Scarabaeus zambesianus.</a> J. Exp. Biol. 206, 1535-1543 (2003).</li>
<li>Greiner, B., Ribi, W. A., Warrant, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Retinal+and+optical+adaptations+for+nocturnal+vision+in+the+halictid+bee+Megalopta+genalis%5BTitle%5D)+AND+%22Cell+Tiss+Res%22%5BJournal%5D)">Retinal and optical adaptations for nocturnal vision in the halictid bee Megalopta genalis.</a> Cell Tiss Res. 316, 377-390 (2004).</li>
<li>Warrant, E. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Nocturnal+vision+and+landmark+orientation+in+a+tropical+halictid+bee%5BTitle%5D)+AND+%22Curr.+Biol%22%5BJournal%5D)">Nocturnal vision and landmark orientation in a tropical halictid bee.</a> Curr. Biol. 14, 1309-1318 (2004).</li>
<li>Lattke, J. E. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aLattke%2C+JE+%22Ants+Standard+Methods+for+Measuring+and+Monitoring+Biodiversity%22">Ants Standard Methods for Measuring and Monitoring Biodiversity</a>. , 155-171 (2000).</li>
<li>Ribi, W. A. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aRibi%2C+WA+%22A+Handbook+in+Biological+Electron+Microscopy%22">A Handbook in Biological Electron Microscopy</a>. , 1-106 (1987).</li>
<li>Narendra, A., Ramirez-Esquivel, F., Ribi, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Compound+eye+and+ocellar+structure+for+walking+and+flying+modes+of+locomotion+in+the+Australian+ant%2C+Camponotus+consobrinus%5BTitle%5D)+AND+%22Sci.+Rep%22%5BJournal%5D)">Compound eye and ocellar structure for walking and flying modes of locomotion in the Australian ant, Camponotus consobrinus.</a> Sci. Rep. 6, 22331(2016).</li>
<li>Narendra, A., Greiner, B., Ribi, W. A., Zeil, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Light+and+dark+adaptation+mechanisms+in+the+compound+eyes+of+Myrmecia+ants+that+occupy+discrete+temporal+niches%5BTitle%5D)+AND+%22J.+Exp.+Biol%22%5BJournal%5D)">Light and dark adaptation mechanisms in the compound eyes of Myrmecia ants that occupy discrete temporal niches.</a> J. Exp. Biol. 219, 2435-2442 (2016).</li>
<li>Ribi, W. A., Zeil, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+visual+system+of+the+Australian+%26%2334%3BRedeye%26%2334%3B+cicada+%28Psaltoda+moerens%29%5BTitle%5D)+AND+%22Arthr.+Struct.+Dev%22%5BJournal%5D)">The visual system of the Australian &#34;Redeye&#34; cicada (Psaltoda moerens).</a> Arthr. Struct. Dev. 44, 574-586 (2015).</li>
<li>Ribi, W. A., Warrant, E. J., Zeil, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+organization+of+honeybee+ocelli%3A+regional+specializations+and+rhabdom+arrangements%5BTitle%5D)+AND+%22Arthr.+Struct.+Dev%22%5BJournal%5D)">The organization of honeybee ocelli: regional specializations and rhabdom arrangements.</a> Arthr. Struct. Dev. 40, 509-520 (2011).</li>
<li>Ribi, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Colour+receptors+in+the+eye+of+the+digger+wasp%2C+Sphex+cognatus+Smith%3A+evaluation+by+selective+adaptation%5BTitle%5D)+AND+%22Cell+Tiss.+Res%22%5BJournal%5D)">Colour receptors in the eye of the digger wasp, Sphex cognatus Smith: evaluation by selective adaptation.</a> Cell Tiss. Res. 195, 471-483 (1978).</li>
<li>Ribi, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ultrastructure+and+migration+of+screening+pigments+in+the+retina+of+Pieris+rapae+L.+%28Lepidoptera%2C+Pieridae%29%5BTitle%5D)+AND+%22Cell+Tiss.+Res%22%5BJournal%5D)">Ultrastructure and migration of screening pigments in the retina of Pieris rapae L. (Lepidoptera, Pieridae).</a> Cell Tiss. Res. 191, 57-73 (1978).</li>
<li>Lau, T., Gross, E., Meyer-Rochow, V. B. <a target="_blank" href="https://www.eje.cz/pdfs/eje/2007/03/14.pdf">Sexual dimorphism and light/dark adaptation in the compound eyes of male and female Acentria ephemerella (Lepidoptera: Pyraloidea: Crambidae).</a> Eur. J. Entomol. 104, 459-470 (2007).</li>
<li>Wipfler, B., Pohl, H., Yavorskaya, M. I., Beutel, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+review+of+methods+for+analysing+insect+structures+-+the+role+of+morphology+in+the+age+of+phylogenomics%5BTitle%5D)+AND+%22Curr.+Opin.+Insect+Sci%22%5BJournal%5D)">A review of methods for analysing insect structures - the role of morphology in the age of phylogenomics.</a> Curr. Opin. Insect Sci. 18, 60-68 (2016).</li>
<li>Streinzer, M., Brockmann, A., Nagaraja, N., Spaethe, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Sex+and+caste-specific+variation+in+compound+eye+morphology+of+five+honeybee+species%5BTitle%5D)+AND+%22PLoS+ONE%22%5BJournal%5D)">Sex and caste-specific variation in compound eye morphology of five honeybee species.</a> PLoS ONE. 8, e57702(2013).</li>
<li>Somanathan, H., Warrant, E. J., Borges, R. M., Wallén, R., Kelber, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Resolution+and+sensitivity+of+the+eyes+of+the+Asian+honeybees+Apis+florea%2C+Apis+cerana+and+Apis+dorsata%5BTitle%5D)+AND+%22J.+Exp.+Biol%22%5BJournal%5D)">Resolution and sensitivity of the eyes of the Asian honeybees Apis florea, Apis cerana and Apis dorsata.</a> J. Exp. Biol. 212, 2448-2453 (2009).</li>
</ol>

著者は競争の興味を宣言しません。

上記の方法のスイート アリなどの昆虫の光学系に効果的な調査を可能にします。これらのテクニックは、サンプリングの解像度、光感度で、検討されている目の潜在的な偏波依存性の私達の理解をお知らせ。この知識は、視覚能力に生理・行動の調査のための重要な基礎を提供します。さらに、ここでの方法は、ant の視覚系に焦点を当てている、これらのテクニック使用できます他の昆虫 (...

ビジョンは、ほとんどの動物のための重要な感覚モダリティです。ビジョンは、正確に示す目標の確立し、ルートに従うこととコンパス情報1,2を取得ためのナビゲーションのコンテキストで特に重要です。昆虫は複眼のペアを使用して視覚情報を検出し、いくつかのケースで 1 ~ 3 背側に配置されたシンプルな目は単眼3,4,5と呼ばれます。
アリの目は、ので、種を渡っていくつかのキーの特性を保全するアリは驚くほど多様なのですが、特に興味深いのです。解剖学、サイズ、および生態系の劇的な変化にもかかわらず種の大半社会性、コロニーに住んでいます。結果として、異なる種中心的な場所とリソース間の前後移動の面で同様の視覚課題に直面します。体長、厳密に夜行性の種に専ら昼間および視覚捕食者6,7、跳躍する地下歩行が遅いから 0.5 〜 26 ミリメートルに至る動物の蟻の間で同じ基本的な目 bauplan を観察できます。 8,9,10。すべての生態および行動のこれらの驚異的な違いは本体サイズ11,12ライフ スタイル、さまざまな環境に合わせて同じ基本的な眼の構造の無数の順列に上昇を与えます。結果として、アリのビジュアルの生態を勉強断固とした探偵に可能性の宝庫を提供します。
昆虫の視覚系を理解することは、行動能力に洞察力を得ることに不可欠です。これは、素敵な生態と、いくつかの昆虫のグループ (例えば、参照13,14,15,16,で大成功を動作解剖学を組み合わせた統合的な研究から明らかです。17). ant ナビゲーションと蟻の行動のフィールドは、一般的にかなり成功している、しかし非常に少し重点を ant ビジョン、いくつか選択した種の外に配置されています。ここでは、私たちは、アリの目のデザインを調査する技術について詳しく説明します。我々 は、アリに集中しながらこれらの技術適用できます、他の昆虫のわずかな変更でも。

<table><tbody><tr><td>Ant</td><td>Myrmecia midas</td><td></td><td></td></tr><tr><td>Stereomicroscope</td><td>Leica M205 FA</td><td></td><td></td></tr><tr><td>Sputter coater</td><td>Pro Sci Tech</td><td></td><td></td></tr><tr><td>Ethanol</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Petri dish</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Dissecting microscope</td><td>Leica MZ6</td><td></td><td></td></tr><tr><td>Insect Pin</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Colourless nail polish</td><td>Non branded: from any cosmetic store</td><td></td><td></td></tr><tr><td>Glass slide</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Razor blade</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Foreceps</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Cover slip</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Compound microscope</td><td>Leica DM5000 B</td><td></td><td></td></tr><tr><td>Glutaraldehyde</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Paraformalydehyde</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Potassium Chloride (KCl)</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>di-Sodium Hydrogen phosphate (Na2HPO4)</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Potassium di-Hydrogen Phosphate (KH2PO4)</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Sodium Chloride (NaCl)</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Osmium tetroxide</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Acetone</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Araldite Epoxy Resin</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Pasteur pipette</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Toluidie Blue</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Hotplate</td><td>Riechert HK120</td><td></td><td></td></tr></tbody></table>

techniques for investigating the anatomy of the ant visual system

この記事は光学顕微鏡 (LM) における技術のスイートについて説明と電子顕微鏡検査 (EM) 昆虫の内部および外部の目の解剖学を研究するために使用できます。アリの目に仕事用に最適化され、透過型電子顕微鏡 (TEM)、走査型電子顕微鏡 (SEM) など他の技術と連携して動作させる従来の組織学的技術が含まれます。これらの技術は、非常に役に立つ、困難な場合が初心者顕微鏡の重視は、トラブルシューティングと異なる標本の最適化に関するこの記事に配置されているので。全体の標本 (写真顕微鏡・ SEM) のイメージングに関する情報を提供し、それぞれの長所と短所を話し合います。我々 は全体の目のレンズの直径を決定する際に使用される技術を強調表示し、改善のための新しい手法を説明します。最後に、LM と TEM の試料を準備にかかわっている技術について述べる断面、染色、およびこれらのサンプル画像します。我々 はサンプルを準備するときに、彼らの周りを移動する最善の方法を来るかもしれない 1 つのハードルをについて説明します。

ここで説明する方法では、単純なものの詳細な研究とアリの複眼を有効にします。Z ただし技術を使用して頭の背表示画像の視覚システム (図 1) レイアウトの概要を取得することができます。これは良い準備解剖、必要な断面の角度を決定します。この手法、また頭の幅、目の長さ、単眼レンズの直径などの測定に役立ちます。SEM イメージング?...

Watch this Scientific Journal Video about Techniques for Investigating the Anatomy of the Ant Visual System at JoVE.com

Techniques for Investigating the Anatomy of the Ant Visual System

この記事では、光と昆虫の内部および外部の目の解剖学を研究、電子顕微鏡の技術のスイートについて説明します。アリの目、詳細なトラブルシューティングと異なる標本や関心領域の最適化のための提案の仕事のために最適化されたいくつかの伝統的なテクニックが含まれます。

Ant の視覚系の解剖学を調査するためのテクニック

This article outlines a suite of techniques in light microscopy (LM) and electron microscopy (EM) which can be used to study the internal and external eye ...

techniques-for-investigating-the-anatomy-of-the-ant-visual-system

Research

JoVE Journal

Environment

13.3K ビュー.  Australian National University. この記事では、光と昆虫の内部および外部の目の解剖学を研究、電子顕微鏡の技術のスイートについて説明します。アリの目、詳細なトラブルシューティングと異なる標本や関心領域の最適化のための提案の仕事のために最適化されたいくつかの伝統的なテクニックが含まれます。

ビデオ: Techniques for Investigating the Anatomy of the Ant Visual System

Visual Classical Conditioning in Wood Ants

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, studies implementing classical conditioning paradigms with harnessed insects have been important for investigating the exact cues that individuals learn and the neural mechanisms underlying learning and memory formation. Here we present a protocol for evoking visual associative learning in wood ants through classical conditioning. In this paradigm, ants are harnessed and presented with a visual cue (a blue cardboard), the conditional stimulus (CS), paired with an appetitive sugar reward, the unconditional stimulus (US). Ants perform a Maxilla-Labium Extension Reflex (MaLER), the unconditional response (UR), which can be used as a readout for learning. Training consists of 10 trials, separated by a 5-minute intertrial interval (ITI). Ants are also tested for memory retention 10 minutes or 1 hour after training. This protocol has the potential to allow researchers to analyze, in a precise and controlled manner, the details of visual memory formation and the neural basis of learning and memory formation in wood ants.

We present a protocol for the classical conditioning of harnessed ants that permits researchers to study visual learning and memory formation at a level of analysis not possible with freely moving individuals.

木蟻で visual の古典的条件付け

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, ...

行動学

The Gateway to the Brain: Dissecting the Primate Eye

The visual system in humans is considered the gateway to the world and plays a principal role in the plethora of sensory, perceptual and cognitive processes. It is therefore not surprising that quality of vision is tied to quality of life . Despite widespread clinical and basic research surrounding the causes of visual disorders, many forms of visual impairments, such as retinitis pigmentosa and macular degeneration, lack effective treatments. Non-human primates have the closest general features of eye development to that of humans. Not only do they have a similar vascular anatomy, but amongst other mammals, primates have the unique characteristic of having a region in the temporal retina specialized for high visual acuity, the fovea1. Here we describe a general technique for dissecting the primate retina to provide tissue for retinal histology, immunohistochemistry, laser capture microdissection, as well as light and electron microscopy. With the extended use of the non-human primate as a translational model, our hope is that improved understanding of the retina will provide insights into effective approaches towards attenuating or reversing the negative impact of visual disorders on the quality of life of affected individuals.

The non-human primate is an important translational species for our understanding of development and aging. The anatomical organization of the primate retina may provide important insights into normal and pathological conditions in humans.

脳へのゲートウェイ：霊長目の解剖

The visual system in humans is considered the  gateway  to the world and plays a principal role in the plethora of sensory, perceptual and cognitive ...

生物学

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees (Apis mellifera L.)

Honey bees (Apis mellifera L.) are eusocial insects and well known for their complex division of labor and associative learning capability1, 2. The worker bees spend the first half of their life inside the dark hive, where they are nursing the larvae or building the regular hexagonal combs for food (e.g. pollen or nectar) and brood3. The antennae are extraordinary multisensory feelers and play a pivotal role in various tactile mediated tasks4, including hive building5 and pattern recognition6. Later in life, each single bee leaves the hive to forage for food. Then a bee has to learn to discriminate profitable food sources, memorize their location, and communicate it to its nest mates7. Bees use different floral signals like colors or odors7, 8, but also tactile cues from the petal surface9 to form multisensory memories of the food source. Under laboratory conditions, bees can be trained in an appetitive learning paradigm to discriminate tactile object features, such as edges or grooves with their antennae10, 11, 12, 13. This learning paradigm is closely related to the classical olfactory conditioning of the proboscis extension response (PER) in harnessed bees14. The advantage of the tactile learning paradigm in the laboratory is the possibility of combining behavioral experiments on learning with various physiological measurements, including the analysis of the antennal movement pattern.

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees Apis mellifera L.

In this protocol we show how to condition harnessed honey bees to tactile stimuli and introduce a 2D motion capture technique for analyzing the kinematics of fine-scale antennal sampling pattern.

蜂蜜の蜂で触角サンプリング戦略の触覚エアコンと運動解析（<em&gt;ミツバチ</em&gt; L.）

Honey bees (Apis mellifera L.) are eusocial insects and well known for their complex division of labor and associative learning capability1, 2. The worker ...

神経科学

Dissection, Immunohistochemistry and Mounting of Larval and Adult Drosophila Brains for Optic Lobe Visualization

The Drosophila optic lobe, comprised of four neuropils: the lamina, medulla, lobula and lobula plate, is an excellent model system for exploring the developmental mechanisms that generate neural diversity and drive circuit assembly. Given its complex three-dimensional organization, analysis of the optic lobe requires that one understand how its adult neuropils and larval progenitors are positioned relative to each other and the central brain. Here, we describe a protocol for the dissection, immunostaining and mounting of larval and adult brains for optic lobe imaging. Special emphasis is placed on the relationship between mounting orientation and the spatial organization of the optic lobe. We describe three mounting strategies in the larva (anterior, posterior and lateral) and three in the adult (anterior, posterior and horizontal), each of which provide an ideal imaging angle for a distinct optic lobe structure.

Dissection, Immunohistochemistry and Mounting of Larval and Adult Drosophila Brains for Optic Lobe Visualization

This protocol describes three steps to prepare larval and adult Drosophila optic lobes for imaging: 1) brain dissections, 2) immunohistochemistry and 3) mounting. Emphasis is placed on step 3, as distinct mounting orientations are required to visualize specific optic lobe structures.&#160;

視葉可視化のためのショ ウジョウバエ の幼虫および成虫の脳の解剖、免疫組織化学、およびマウント

The Drosophila optic lobe, comprised of four neuropils: the lamina, medulla, lobula and lobula plate, is an excellent model system for exploring the ...

発生生物学

An Explant System for Time-Lapse Imaging Studies of Olfactory Circuit Assembly in Drosophila

~Neurons are precisely interconnected to form circuits essential for the proper function of the brain. The Drosophila olfactory system provides an excellent model to investigate this process since 50 types of olfactory receptor neurons (ORNs) from the antennae and maxillary palps project their axons to 50 identifiable glomeruli in the antennal lobe and form synaptic connections with dendrites from 50 types of second-order projection neurons (PNs). Previous studies mainly focused on identifying important molecules that regulate the precise targeting in the olfactory circuit using fixed tissues. Here, an antennae-brain explant system that recapitulates key developmental milestones of olfactory circuit assembly in culture is described. Through dissecting the external cuticle and cleaning opaque fat bodies covering the developing pupal brain, high quality images of single neurons from live brains can be collected using two-photon microscopy. This allows time-lapse imaging of single ORN axon targeting from live tissue. This approach will help reveal important cell biological contexts and functions of previously identified important genes and identify mechanisms underpinning the dynamic process of circuit assembly.

An Explant System for Time-Lapse Imaging Studies of Olfactory Circuit Assembly in Drosophila

This protocol describes the dissection procedure, culture condition, and live imaging of an antennae-brain explant system for the study of the olfactory circuit assembly.

ショウジョウバエの嗅覚回路アセンブリのタイムラプスイメージング研究のための外植システム

~Neurons are precisely interconnected to form circuits essential for the proper function of the brain. The Drosophila olfactory system provides an ...

Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light

The nervous system has the remarkable ability to adapt and respond to various stimuli. This neural adjustment is largely achieved through plasticity at the synaptic level. The Active Zone (AZ) is the region at the presynaptic membrane that mediates neurotransmitter release and is composed of a dense collection of scaffold proteins. AZs of Drosophila melanogaster (Drosophila) photoreceptors undergo molecular remodeling after prolonged exposure to natural ambient light. Thus the level of neuronal activity can rearrange the molecular composition of the AZ and contribute to the regulation of the functional output.
Starting from the light exposure set-up preparation to the immunohistochemistry, this protocol details how to quantify the number, the spatial distribution, and the delocalization level of synaptic molecules at AZs in Drosophila photoreceptors. Using image analysis software, clusters of the GFP-fused AZ component Bruchpilot were identified for each R8 photoreceptor (R8) axon terminal. Detected Bruchpilot spots were automatically assigned to individual R8 axons. To calculate the distribution of spot frequency along the axon, we implemented a customized software plugin. Each axon&#39;s start-point and end-point were manually defined and the position of each Bruchpilot spot was projected onto the connecting line between start and end-point. Besides the number of Bruchpilot clusters, we also quantified the delocalization level of Bruchpilot-GFP within the clusters. These measurements reflect in detail the spatially resolved synaptic dynamics in a single neuron under different environmental conditions to stimuli.

Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light

Here we show how to quantify the number and spatial distribution of synaptic active zones in Drosophila melanogaster&#160;photoreceptors, highlighted with a genetically encoded molecular marker, and their modulation after prolonged exposure to light.

のシナプス変調を分析<em&gt;キイロショウジョウバエ</em&gt;長時間の光照射後の感光体

The nervous system has the remarkable ability to adapt and respond to various stimuli. This neural adjustment is largely achieved through plasticity at ...

Analyzing Dendritic Morphology in Columns and Layers

In many regions of the central nervous systems, such as the fly optic lobes and the vertebrate cortex, synaptic circuits are organized in layers and columns to facilitate brain wiring during development and information processing in developed animals. Postsynaptic neurons elaborate dendrites in type-specific patterns in specific layers to synapse with appropriate presynaptic terminals. The fly medulla neuropil is composed of 10 layers and about 750 columns; each column is innervated by dendrites of over 38 types of medulla neurons, which match with the axonal terminals of some 7 types of afferents in a type-specific fashion. This report details the procedures to image and analyze dendrites of medulla neurons. The workflow includes three sections: (i) the dual-view imaging section combines two confocal image stacks collected at orthogonal orientations into a high-resolution 3D image of dendrites; (ii) the dendrite tracing and registration section traces dendritic arbors in 3D and registers dendritic traces to the reference column array; (iii) the dendritic analysis section analyzes dendritic patterns with respect to columns and layers, including layer-specific termination and planar projection direction of dendritic arbors, and derives estimates of dendritic branching and termination frequencies. The protocols utilize custom plugins built on the open-source MIPAV (Medical Imaging Processing, Analysis, and Visualization) platform and custom toolboxes in the matrix laboratory language. Together, these protocols provide a complete workflow to analyze the dendritic routing of Drosophila medulla neurons in layers and columns, to identify cell types, and to determine defects in mutants.

Here, we show how to analyze dendritic routing of Drosophila medulla neurons in columns and layers. The workflow includes a dual-view imaging technique to improve the image quality and computational tools for tracing, registering dendritic arbors to the reference column array and for analyzing the dendritic structures in 3D space.

列およびレイヤーで樹状形態を分析します

In many regions of the central nervous systems, such as the fly optic lobes and the vertebrate cortex, synaptic circuits are organized in layers and ...

<meta charset="utf8"></head><body>ショウジョウバエの若年脳における嗅覚体験誘発性シナプスリモデリング

Experience-Dependent Remodeling of Juvenile Brain Olfactory Sensory Neuron Synaptic Connectivity in an Early-Life Critical Period

Early-life olfactory sensory experience induces dramatic synaptic glomeruli remodeling in the Drosophila juvenile brain, which is experientially dose-dependent, temporally restricted, and transiently reversible only in a short, well-defined critical period. The directionality of brain circuit synaptic connectivity remodeling is determined by the specific odorant acting on the respondent receptor class of olfactory sensory neurons. In general, each neuron class expresses only a single odorant receptor and innervates a single olfactory synaptic glomerulus. In the Drosophila genetic model, the full array of olfactory glomeruli has been precisely mapped by odorant responsiveness and behavioral output. Ethyl butyrate (EB) odorant activates Or42a receptor neurons innervating the VM7 glomerulus. During the early-life critical period, EB experience drives dose-dependent synapse elimination in the Or42a olfactory sensory neurons. Timed periods of dosed EB odorant exposure allow investigation of experience-dependent circuit connectivity pruning in juvenile brain. Confocal microscopy imaging of antennal lobe synaptic glomeruli is done with Or42a receptor-driven transgenic markers that provide quantification of synapse number and innervation volume. The sophisticated Drosophila genetic toolkit enables the systematic dissection of the cellular and molecular mechanisms mediating brain circuit remodeling.

<meta charset="utf8"></head><body>著者スポットライト:臨界期の経験依存性シナプス刈り込みにおけるグリアの影響の調査

We describe here methods for inducing and analyzing olfactory experience-dependent remodeling of antennal lobe synaptic glomeruli in the Drosophila juvenile brain.

Ant の視覚系の解剖学を調査するためのテクニック

要約

さらに動画を探す

木蟻で visual の古典的条件付け

脳へのゲートウェイ：霊長目の解剖

蜂蜜の蜂で触角サンプリング戦略の触覚エアコンと運動解析（

視葉可視化のためのショウジョウバエの幼虫および成虫の脳の解剖、免疫組織化学、およびマウント

ショウジョウバエの嗅覚回路アセンブリのタイムラプスイメージング研究のための外植システム

のシナプス変調を分析

列およびレイヤーで樹状形態を分析します

若年脳嗅覚神経細胞シナプス結合性の経験依存的リモデリング

若年脳嗅覚神経細胞シナプス結合性の経験依存的リモデリング

若年脳嗅覚神経細胞シナプス結合性の経験依存的リモデリング

Ant の視覚系の解剖学を調査するためのテクニック

要約

さらに動画を探す

木蟻で visual の古典的条件付け

脳へのゲートウェイ：霊長目の解剖

蜂蜜の蜂で触角サンプリング戦略の触覚エアコンと運動解析（

視葉可視化のためのショ ウジョウバエ の幼虫および成虫の脳の解剖、免疫組織化学、およびマウント

ショウジョウバエの嗅覚回路アセンブリのタイムラプスイメージング研究のための外植システム

のシナプス変調を分析

列およびレイヤーで樹状形態を分析します

若年脳嗅覚神経細胞シナプス結合性の経験依存的リモデリング

若年脳嗅覚神経細胞シナプス結合性の経験依存的リモデリング

若年脳嗅覚神経細胞シナプス結合性の経験依存的リモデリング

視葉可視化のためのショウジョウバエの幼虫および成虫の脳の解剖、免疫組織化学、およびマウント