著者はE.を供給するための6月ミドルトンに感謝したいと思いますフェカリス株と指導のための細菌文化を持つ。C.エレガンスは、研究基盤プログラムのNIH事務所（P40 OD010440）によって資金を供給されるCGC、によって提供された。

1）線虫の増殖培地の調製（NGM） <ol><li> 50プレートを作成するには、1.5グラムのNaCl、8.5グラム超純寒天、及び1 Lflask〜1.25グラムペプトンを追加します。 </li><li>フラスコにのdH 2 Oの487.5ミリリットルを追加します。アルミホイルの切れ端でフラスコの開口部を混合し、カバーするために静かに渦巻き。 </li><li>標準的な条件（121 PSI、120°C、20分）でオートクレーブして、ソリューションを滅菌。 </li><li>解決策は、水浴中で45℃に冷却することができます。 </li><li> 5 mg / mlのストック溶液（エタノールに溶解）、500μlの1 M CaCl 2を、500μlの1 M MgSO 4を、そして12.5ミリリットルKPO 4緩衝液（54.15グラムから500μlのコレステロール：溶液に、順番に次の行を追加します。のdH 2 Oを500mlの中でKH 2 PO 4および17.8グラムK 2 HPO 4）18に記載されているように。 </li><li>滅菌ピペットを使用して、各ポリスチレンを60mmペトリ皿に寒天溶液10mlを加える。メディアは室温overnで固化することを許可ightの。 </li></ol> BブロスE. 2）準備大腸菌メディア<ol><li> 5を100mlの培養液を作るために、ビーカーにバクトトリプトン、2.5グラムの酵母エキス、5.0グラムのNaCl 5.0グラムを追加します。 </li><li> 500mlを最終容量まで無菌蒸留水を加える。溶質が溶解するまで、撹拌しながら、ホットプレート上でソリューションを加熱。 </li><li> 5ガラス瓶（少なくとも200ミリリットルの容積）にBブイヨンを100mlを注ぐ。 </li><li>標準的な条件（121 PSI、120℃、20分）でオートクレーブで解決策を滅菌する。 </li><li> B液は使用前に室温まで冷却することができます。 </li></ol> 3）種まきC.エレガンスメンテナンスプレート<ol><li> NGMプレートは少なくとも1週間室温で乾燥させた後、E.のいくつかのコロニー〜と100ミリリットルBのブロス培養物を接種する大腸菌 （OP50）。 </li><li> E.を育てる37℃で一晩大腸菌文化。 </li><li>ピペットワンツー滴（約10081、L）OP50文化の各NGMプレートの中央に。プレートは、典型的には、蓋側を上に積層されている。 （播種）ピペッティングと、スタックの一番下にあるプレートに文化をピペッティングすることから始めます。各スタックの上のあなたのように動作します。文化は、プレートの端に近づきすぎていないことが重要であるので、播種後のプレートを移動しないようにしてください。 </li><li>シードプレートは使用する前に、少なくとも1週間、室温で乾燥させます。 </li><li>シードされたプレートを室温で密封されたプラスチック容器に（反転）が格納される。 </li></ol> 4）C.を維持エレガンス菌株<ol><li>よく供給C.を維持するためにelegansの株は、3〜4日ごとに新しい、シードNGMプレートに3 L4sまたは妊娠成人を移動するピックワームを使用しています。 </li><li>ストアプレート（反転）は、好ましくはインキュベーター内部15℃ - 22℃記載のアッセイにおける文化20℃で保存した後elegansの開発は、温度依存性である。 maintenaとして温度が上昇NCE、発達段階の間の時間は減少します。 </li></ol>トリプシン大豆寒天（TSA）Eの5）の調製フェカリス文化メディア<ol><li>ホットプレート上で、攪拌しながら50のプレートを作るために、滅菌したフラスコに、500mlの蒸留水を測定し、沸騰させる。 </li><li>フラスコに粉末TSA寒天20gのを加え、溶液を溶解することができます。 </li><li>標準的な条件（121 PSI、120℃、20分）でオートクレーブで解決策を滅菌する。 </li><li>解決策は、水浴中で45℃に冷却することができます。 </li><li>滅菌ピペットを使用して、各ポリスチレンを60mmペトリ皿に寒天溶液10mlを加える。メディアが一晩室温で固化することができます。 </li></ol>トリプシン大豆ブロス（TSB）Eの6）の準備フェカリス文化メディア<ol><li>攪拌しながら250培養管を作るために、ホットプレート上でフラスコと暖かいに滅菌蒸留水500mlを測定します。 </lI&gt; <li>フラスコにTSB寒天粉末15グラムを加え、粉を溶解することができます。 </li><li>標準的な条件（121 PSI、120℃、20分）でオートクレーブで解決策を滅菌する。 </li><li>滅菌、5ミリリットル試験管にその後ピペット2ミリリットル、室温まで冷却するソリューションを可能にします。 </li></ol> 7）ストレプトマイシンと、E.のためのブレインハートインフュージョン（BHI）メディアの準備、 フェカリス文化<ol><li>滅菌フラスコに、500mlの蒸留水を測定し、ホットプレート上で、攪拌しながら、沸騰させる。 </li><li>フラスコにBHI培地の26グラムを加え、溶液を溶解することができます。 </li><li>標準的な条件（121 PSI、120℃、20分）でオートクレーブで解決策を滅菌する。 </li><li>ソリューションは℃の水浴中で45まで冷まします。 </li><li>無菌技術を使用して、10 mg / mlのストレプトマイシンストック溶液と混合する渦0.5 mlを加える。 </li><li>各60ミリメートルのポリスチレンシャーレに溶液10mlを加える。 </li><l私は&gt;メディアが一晩室温で固化することを許可する。ストアプレート（反転）4℃で使用前に数時間までC。 </li></ol> 8）E.を維持フェカリス菌株<ol><li> E.を維持するフェカリス株は、独立したトリプシン大豆寒天（TSA）プレートにストリーク個々のコロニーを滅菌ループを使用しています。 </li><li> 37℃で一晩培養液を育てる。数週間、室温で密封された袋や箱に保管プレート（反転）。 </li></ol> 9）E.の準備EIWアッセイ用フェカリスプレート<ol><li> E.のコロニーを2ミリリットルTSB文化を接種フェカリス 、37℃で一晩インキュベート℃、 </li><li> E.のピペットを20μl フェカリス準備BHI-ストレプトマイシンプレートの中央に文化、そして37℃で一晩インキュベート℃、 </li></ol> 10）E.の準備EIWアッセイ用大腸菌プレート（コントロールプレート） <ol><li>の100ml B-仲間に接種E.の植民地と目文化大腸菌 OP50と37℃で一晩インキュベート</li><li>非シードNGMプレートの中央にOP50文化のピペットを20μlと播いプレートは室温で一晩乾燥させます。 </li></ol>ワームアッセイ11）卵<ol><li> 15ピック- 20上演-L4 C.準備BHI-ストレプトE.上の細菌の芝生の中央に虫 （ 図1） フェカリスプレート 。 BHIプレートにOP50の多くを転送しないように注意してください。 NGM-OP50プレートコントロールにL4s同じ数を選ぶ。 </li><li> 20℃で40時間、プレートをインキュベート</li><li> 20％漂白剤溶液を準備します。蒸留水の適切な量と市販漂白剤（6.0％次亜塩素酸ナトリウム）を混ぜる。 </li><li>プラスチックの蓋の上に10の個別の場所（ワームまたは細菌プレート）に漂白剤溶液10μl滴を追加します。 </li><li>ワームピックを使用して、各漂白ドロップに1ワームを転送します。静かに渦の先端ワームは、ピックの先端を（解剖顕微鏡下で滴を表示することによって確認）洗い流されていることを確認するために漂白剤の液滴で選ぶ。 </li><li>ワームは卵を（ 図2）追放、オープンバーストまでキューティクルが約10分間、または溶解することができます。プレートから卵を転送しないように注意してください。 </li><li>解剖顕微鏡を使用して、漂白剤の各ドロップで卵を数える。 </li></ol>

<ol>
	<li>Horvitz, H. R., Chalfie, M., Trent, C., Sulston, J. E., Evans, P. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serotonin+and+octopamine+in+the+nematode+Caenorhabditis+elegans.">Serotonin and octopamine in the nematode Caenorhabditis elegans.</a> Science. 216, 1012-1014 (1982).</li><li>Sawin, E. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Genetic and cellular analysis of modulated behaviors in Caenorhabditis elegans [dissertation]. , (1996).</li><li>Trent, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Genetic and behavioral studies of the egg-laying system of Caenorhabditis elegans [dissertation]. , (1982).</li><li>Chase, D. L., Koelle, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+analysis+of+RGS+protein+function+in+Caenorhabditis+elegans.">Genetic analysis of RGS protein function in Caenorhabditis elegans.</a> Methods Enzymol. 389, 305-320 (2004).</li><li>Trent, C., Tsuing, N., Horvitz, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Egg-laying+defective+mutants+of+the+nematode+Caenorhabditis+elegans.">Egg-laying defective mutants of the nematode Caenorhabditis elegans.</a> Genetics. 104, 619-647 (1983).</li><li>Weinshenker, D., Garriga, G., Thomas, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+and+pharmacological+analysis+of+neurotransmitters+controlling+egg+laying+in+Caenorhabditis+elegans.">Genetic and pharmacological analysis of neurotransmitters controlling egg laying in Caenorhabditis elegans.</a> J. Neurosci. 15, 6975-6985 (1995).</li><li>Waggoner, L. E., Zhou, G. T., Schafer, R. W., Schafer, W. R. <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+alternative+behavioral+states+by+serotonin+in+Caenorhabditis+elegans.">Control of alternative behavioral states by serotonin in Caenorhabditis elegans.</a> Neuron. 21, 203-214 (1998).</li><li>Bany, A., Dong, M., Koelle, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+and+cellular+basis+for+acetylcholine+inhibition+of+Caenorhabditis+elegans+egg-laying+behavior.">Genetic and cellular basis for acetylcholine inhibition of Caenorhabditis elegans egg-laying behavior.</a> J. Neurosci. 23, 8060-8069 (2003).</li><li>Dempsey, C. M., Mackenzie, S. M., Gargus, A., Blanco, G., Sze, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serotonin+(5HT),+fluoxetine,+imipramine+and+dopamine+target+distinct+5HT+receptor+signaling+to+modulate+Caenorhabditis+elegans+egg-laying+behavior.">Serotonin (5HT), fluoxetine, imipramine and dopamine target distinct 5HT receptor signaling to modulate Caenorhabditis elegans egg-laying behavior.</a> Genetics. 169, 1425-1436 (2005).</li><li>Leung, M. C., Williams, P. L., Bennedetto, A., Au, C., Helmcke, K. J., Aschner, M., Meyer, 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=Caenorhabditis+elegans:+an+emerging+model+in+biomedical+and+environmental+toxicology.">Caenorhabditis elegans: an emerging model in biomedical and environmental toxicology.</a> Toxicol. Sci. 106, 5-28 (2008).</li><li>Mylonakis, E., Aballay, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Worms+and+flies+as+genetically+tractable+animal+models+to+study+host-pathogen+interactions.">Worms and flies as genetically tractable animal models to study host-pathogen interactions.</a> Infect. Immun. 73, 3833-3841 (2005).</li><li>Schafer, W. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> WormBook. , (2005).</li><li>White, J., Southgate, E., Thomson, N., Brenner, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+structure+of+the+Caenorhabditis+elegans+nervous+system.">The structure of the Caenorhabditis elegans nervous system.</a> Philos. Trans. R. Soc. Lond. B. Biol. Sci. 314, 1-340 (1986).</li><li>Desai, C., Garriga, G., McIntire, S. L., Horvitz, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+genetic+pathway+for+the+development+of+the+Caenorhabditis+elegans+HSN+motor+neurons.">A genetic pathway for the development of the Caenorhabditis elegans HSN motor neurons.</a> Nature. 336, 638-646 (1988).</li><li>Garsin, D. A., Sifri, C. D., Mylonakis, E., Qin, X., Singh, K. V., Murray, B. E., Calderwood, S. B., Ausubel, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+model+host+for+identifying+Gram-positive+virulence+factors.">A simple model host for identifying Gram-positive virulence factors.</a> Proc. Natl. Acad. Sci. U.S.A. 98, 10892-10897 (2001).</li><li>Aballay, A., Yorgey, P., Ausubel, F. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Salmonella+typhmurium+proliferates+and+establishes+a+persistent+infection+in+the+intestine+of+Caenorhabditis+elegans.">Salmonella typhmurium proliferates and establishes a persistent infection in the intestine of Caenorhabditis elegans.</a> Curr. Biol. 10, 1539-1542 (2000).</li><li>O'Quinn, A. L., Wiegand, E. M., Jeddeloh, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burkholderia+pseudomallei+kills+the+nematode+Caenorhabditis+elegans+using+an+endotoxin-mediated+paralysis.">Burkholderia pseudomallei kills the nematode Caenorhabditis elegans using an endotoxin-mediated paralysis.</a> Cell. Microbiol. 3, 381-393 (2001).</li><li>Brenner, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetics+of+Caenorhabditis+elegans.">Genetics of Caenorhabditis elegans.</a> Genetics. 77, 71-94 (1974).</li><li>Harvey, S. C., Shorto, A., Orbidans, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+genetic+analysis+of+life-+history+traits+of+Caenorhabditis+elegans+in+stressful+environments.">Quantitative genetic analysis of life- history traits of Caenorhabditis elegans in stressful environments.</a> BMC Evol. Biol. 8, 15 (2008).</li><li>Harvey, S. C., Orbidans, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=All+eggs+are+not+equal:+the+maternal+environment+affects+progeny+reproduction+and+developmental+fate+in+Caenorhabditis+elegans.">All eggs are not equal: the maternal environment affects progeny reproduction and developmental fate in Caenorhabditis elegans.</a> PLoS One. 6, e25840 (2011).</li><li>Klass MR, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aging+in+the+nematode+Caenorhabditis+elegans:+major+biological+and+environmental+factors+influencing+life+span.">Aging in the nematode Caenorhabditis elegans: major biological and environmental factors influencing life span.</a> Mech. Ageing. Dev. 6, 413-429 (1977).</li></ol>

著者は、彼らが競合する経済的利益を持っていないことを宣言します。

正常にこのアッセイを実施する中で最も重要なステップは次のとおりである：1）C.のよく供給銘柄を使用して正確に識別虫 、2）アッセイプレート上の細菌の培養、単一種類、3）E.への露出のためにL4ワームを上演フェカリス、4）Eに露光時間を維持するすべての試験および5にわたって一貫フェカリス ）漂白時間は卵の崩壊を防ぐために、10分を超えてはなら...

線虫（Caenorhabditis elegans）、顕微鏡、自由生活回虫は、伝統的に人間に、その透明な解剖学、十分に特徴付け開発、完全に順ゲノム、短い世代時間、および遺伝的相同性の発達と細胞シグナル伝達過程を研究するために使用されるモデル生物である。より最近では、C.エレガンスは、環境毒物学と先天性免疫10、11の分野でモデル生物となっています。 これらの自己受精雌雄同体のワームは卵から孵化2〜3日以内に、性的に成熟したとなります。そのライフサイクル、C.中エレガンスは、成人に達する前に、4幼虫（L1〜L4）を通過する。 One孤立ふたなり、ピーク産卵から3日以内に、平均して、300子孫を生成することができます。生殖成熟したCでエレガンス雌雄同体は、受精卵が敷設される前に数時間のために子宮内に保持されています。ザ任意の一時点で子宮内に格納された卵の数は、通常の（ピーク産卵時）10及び15 12の間である。子宮内の卵の数は、産卵率と産卵率の両方の関数である。受精卵は、外陰部13の開口部の周囲に配置された16外陰部の筋肉の収縮によって子宮から追放されています。ふたなり特有の運動ニューロン（HSNの）とVCの運動ニューロンのシナプス筋収縮に影響を与える外陰部の筋肉の上に、したがって、産卵行動5,7,13,14。子宮から卵の追放は、ニューロンと筋肉の協調活動に起因して発生する。 C.のラボ文化エレガンスは 、通常非病原大腸菌 OP50の食事で育てています。自然環境、Cの線虫は、例えば病原性細菌などの食品、さまざまなソースに接触し、それは潜在的に有害であり得る。中の有害物質にさらされた場合環境、C.環境がより有利になるまで虫卵を保持します。おそらく、この卵の保持はその子孫を守るための努力です。 ワーム（EIW）アッセイでは、この卵で、C.エレガンスは 、潜在的に病原性細菌、環境で発見されている腸球菌を 、公開されています。 E.の病原フォームへの暴露フェカリスはCで永続的な腸の感染症や死を引き起こす可能性がありますエレガンス 15。病原性細菌の他の形態への曝露は、しかし、効果を定量されなかった、卵の保持16,17に影響を及ぼすことが示されている。また、E.の軽度の病原性株の効果産卵行動に関するフェカリス 、すぐに致死でない株は、研究されていない。 EIWアッセイは、C.の子宮内に保持された卵の数を数える含むエレガンス 4。にもかかわらずC.エレガンス透明であり、子宮内に蓄積した卵は、無傷の動物で定量化することは困難になる可能性があります。 EIWアッセイは妊娠成人Cを漂白関与一定期間細菌に曝露した虫 。漂白剤溶液は、背後に卵を残して外側のキューティクルを溶解する。卵は、保護卵殻の存在による漂白の効果に屈折率である。漂白した後、一方は非常に簡単に漂白時にワームの子宮から放出された卵の数をカウントすることができる。 記載されたアッセイは、一度に子宮に卵の数を定量化する、単純で安価かつ迅速な方法であるため、E.の影響を定量化卵の保持にフェカリス 。このアッセイは、卵の保持に対する細菌、環境毒素又は薬剤の他の種類の効果を定量化するために使用することができる。このアッセイはまた、病原性細菌のためのスクリーンとして使用される可能性を秘めている。

<table border="0" cellpadding="0" cellspacing="0" width="2472">
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		<col />
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		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Agar, ultrapure</td>
			<td style="width:191px;">Affymetrix</td>
			<td style="width:119px;">10906</td>
			<td style="width:1791px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Bacto Peptone</td>
			<td style="width:191px;">Becton Dickinson</td>
			<td style="width:119px;">211677</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Bacto Tryptone</td>
			<td style="width:191px;">Becton Dickinson</td>
			<td style="width:119px;">211705</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Brain Heart Infusion dehydrated medium</td>
			<td style="width:191px;">Carolina Biological Supply</td>
			<td style="width:119px;">781781</td>
			<td></td>
		</tr>
		<tr height="18">
			<td height="18" style="height:19px;width:371px;">C. elegans, N2 strain</td>
			<td style="width:191px;">Caenorhabditis Genetics Center</td>
			<td style="width:119px;"></td>
			<td>http://www.cbs.umn.edu/cgc</td>
		</tr>
		<tr height="18">
			<td height="18" style="height:19px;width:371px;">Cholesterol</td>
			<td style="width:191px;">Alfa Aesar</td>
			<td style="width:119px;">A11470</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Culture plates for C. elegans</td>
			<td style="width:191px;">Tritech Research Inc.</td>
			<td style="width:119px;">T3308</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:371px;">Culture plates for E. faecalis</td>
			<td style="width:191px;">Fisher Scientific-Fisherbrand</td>
			<td style="width:119px;">875713</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:371px;">E. coli (OP50)</td>
			<td style="width:191px;">Caenorhabditis Genetics Center</td>
			<td style="width:119px;"></td>
			<td>http://www.cbs.umn.edu/cgc</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">E. faecalis strains</td>
			<td style="width:191px;"></td>
			<td style="width:119px;"></td>
			<td>provided by J. Middleton. All isolates were confirmed as enterococci</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:191px;"></td>
			<td style="width:119px;"></td>
			<td>by observing growth on enterococcosel agar (BBL) and in 6% NaCl broth;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:191px;"></td>
			<td style="width:119px;"></td>
			<td>
			all strains grew at 44.5 &#186;C and were catalase negative and hydrolyzed esculin. A simplified&#160;dichotomous key based on pigmentation and fermentation reactions for six sugars&#160;(arabinose, mannitol, methyl-&#945;-D-glucopyranoside (MGP), ribose, sorbose and sorbitol) allowed&#160;presumptive identification of all E. faecalis strains (Efs lacks pigmentation and is arabinose, MGP&#160;and sorbose negative and sorbitol, mannitol and ribose positive). All presumptive Efs strains&#160;were confirmed using the API 20 STREP system (Biomerieux).
			</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:191px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:191px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:191px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:191px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:191px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Microscope</td>
			<td style="width:191px;">Motic</td>
			<td style="width:119px;">SMZ 168B</td>
			<td>any microscope with transmitted illumination and 50X magnification should be sufficient</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Streptomycin sulfate</td>
			<td style="width:191px;">Fisher BioReagents</td>
			<td style="width:119px;">BP910-50</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:371px;">Tryptic Soy Agar (Soybean-Casein Digest Agar Medium), Difco</td>
			<td style="width:191px;">Becton Dickinson</td>
			<td style="width:119px;">236950</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:371px;">Trypticase Soy Broth (Soybean-Casein Digest Medium), BBL</td>
			<td style="width:191px;">Becton Dickinson</td>
			<td style="width:119px;">211768</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Yeast extract</td>
			<td style="width:191px;">Acros</td>
			<td style="width:119px;">61180-1000</td>
			<td></td>
		</tr>
	</tbody>
</table>

measuring the effects of bacteria on c. elegans behavior using an egg retention assay

C. elegansの産卵行動は、浸透圧1と振動2などの環境手がかりに影響されます。食品Cの合計がない場合に虫も産卵を中止し、その子宮3で受精卵を保持します。しかし、特に異なる食品の源、特に病原性細菌と産卵行動に腸球菌 、の効果は十分に特徴付けされていません。卵内ワーム（EIW）アッセイは、この場合にはE.、細菌の異なる種類の効果を定量化するための有用なツールであるフェカリス、産卵行動に。 EIWアッセイは、C.の子宮内に保持された卵の数を数える含むエレガンス 4。 EIWアッセイは上演漂白、妊娠成人C.を伴うelegansのキューティクルを除去し、動物から保持卵を分離する。前の漂白に、ワームは、細菌（または環境キューの任意のタイプにさらされている）一定の期間。漂白した後、一方は非常に簡単にワームの子宮内部に保持された卵の数をカウントすることができる。 E.後の卵の保存では、このアッセイでは、定量化可能な増加フェカリス露光を容易に測定することができる。 EIWアッセイは、潜在的に病原性の細菌や環境毒素の存在についてスクリーニングするために使用され得る行動アッセイである。また、EIWアッセイは産卵行動がこのようなセロトニンやアセチルコリンなどの神経伝達物質5-9によって変調であるため、シグナル伝達物質に影響を与える薬をスクリーニングするためのツールかもしれません。

このアッセイは、一つC.内に保持された卵の数を定量化することができE.への曝露後のelegansフェカリス 。 L4は、ワームは（彼らの外陰部、 図1上記の透明オープンスペースが特徴）E.にさらされた上演成人期を通じてフェカリス 。 E.への暴露の40時間後フェカリスは 、漂白を行った。キューティクルは、漂白液滴で崩壊したように、...

Watch this Scientific Journal Video about Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay at JoVE.com

Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay | Biology | Video

Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

卵·イン·ワーム（EIW）アッセイは、産卵行動を定量化するための有用な方法である。産卵の変化は、モデル生物の行動反応することができます<em&gt;線虫（Caenorhabditis elegans）</em&gt;このような病原性細菌によって生成されるものとして、潜在的に有害な環境物質へ。

上の細菌の影響を測定する<em&gt; C.エレガンス</em卵保持アッセイを用いて&gt;行動

C. elegans egg-laying behavior is  affected by environmental cues such as osmolarity1 and  vibration2. In the total absence of food C. elegans also cease ...

measuring-effects-bacteria-on-c-elegans-behavior-using-an-egg

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Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

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16.9K ビュー.  Fairleigh Dickinson University. 卵·イン·ワーム（EIW）アッセイは、産卵行動を定量化するための有用な方法である。産卵の変化は、モデル生物の行動反応することができます線虫（Caenorhabditis elegans）このような病原性細菌によって生成されるものとして、潜在的に有害な環境物質へ。

ビデオ: Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

Generation of Stable Transgenic C. elegans Using Microinjection

Transgenic Caenorhabditis elegans can be readily created via microinjection of a DNA plasmid solution into the gonad 1. The plasmid DNA rearranges to form extrachromosomal concatamers that are stably inherited, though not with the same efficiency as actual chromosomes 2. A gene of interest is co-injected with an obvious phenotypic marker, such as rol-6 or GFP, to allow selection of transgenic animals under a dissecting microscope. The exogenous gene may be expressed from its native promoter for cellular localization studies. Alternatively, the transgene can be driven by a different tissue-specific promoter to assess the role of the gene product in that particular cell or tissue. This technique efficiently drives gene expression in all tissues of C. elegans except for the germline or early embryo 3. Creation of transgenic animals is widely utilized for a range of experimental paradigms. This video demonstrates the microinjection procedure to generate transgenic worms. Furthermore, selection and maintenance of stable transgenic C. elegans lines is described.

This video demonstrates the technique of microinjection into the gonad of C. elegans to create transgenic animals.

マイクロインジェクションを使用して安定したトランスジェニック線虫の世代

Transgenic Caenorhabditis elegans can be readily created via microinjection of a DNA plasmid solution into the gonad 1.  The plasmid DNA rearranges to ...

生物学

Measuring Caenorhabditis elegans Life Span on Solid Media

Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The nematode Caenorhabditis elegans has emerged as a principal model used to study the biology of aging. Because virtually every biological subsystem undergoes functional decline with increasing age, life span is the primary endpoint of interest when considering total rate of aging. In nematodes, life span is typically defined as the number of days an animal remains responsive to external stimuli. Nematodes can be propagated either in liquid media or on solid media in plates, and techniques have been developed for measuring life span under both conditions. Here we present a generalized protocol for measuring life span of nematodes maintained on solid nematode growth media and fed a diet of UV-killed bacteria. These procedures can easily be adapted to assay life span under various common conditions, including a diet consisting of live bacteria, dietary restriction, and RNA interference.

Measuring Caenorhabditis elegans Life Span on Solid Media

In this article we present a general protocol for measuring life span of nematodes maintained on solid media with UV-killed bacterial food.

計測線虫（Caenorhabditis elegans）</em固体培地上>寿命

Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The nematode ...

Imaging C. elegans Embryos using an Epifluorescent Microscope and Open Source Software

Cellular processes, such as chromosome assembly, segregation and cytokinesis,are inherently dynamic. Time-lapse imaging of living cells, using fluorescent-labeled reporter proteins or differential interference contrast (DIC) microscopy, allows for the examination of the temporal progression of these dynamic events which is otherwise inferred from analysis of fixed samples1,2. Moreover, the study of the developmental regulations of cellular processes necessitates conducting time-lapse experiments on an intact organism during development. The Caenorhabiditis elegans embryo is light-transparent and has a rapid, invariant developmental program with a known cell lineage3, thus providing an ideal experiment model for studying questions in cell biology4,5and development6-9. C. elegans is amendable to genetic manipulation by forward genetics (based on random mutagenesis10,11) and reverse genetics to target specific genes (based on RNAi-mediated interference and targeted mutagenesis12-15). In addition, transgenic animals can be readily created to express fluorescently tagged proteins or reporters16,17. These traits combine to make it easy to identify the genetic pathways regulating fundamental cellular and developmental processes in vivo18-21. In this protocol we present methods for live imaging of C. elegans embryos using DIC optics or GFP fluorescence on a compound epifluorescent microscope. We demonstrate the ease with which readily available microscopes, typically used for fixed sample imaging, can also be applied for time-lapse analysis using open-source software to automate the imaging process.

Imaging C. elegans Embryos using an Epifluorescent Microscope and Open Source Software

The C. elegans embryo is a powerful system for studying cell biology and development. We present a protocol for live imaging of C. elegans embryos utilizing DIC optics or fluorescence using readily available epifluorescent microscopes and open-source software.

イメージング C.エレガンス</em落射蛍光顕微鏡とオープンソースソフトウェアを使用して>胚

Cellular processes, such as chromosome assembly, segregation and cytokinesis,are inherently dynamic. Time-lapse imaging of living cells, using ...

Visualization of Mitochondrial Respiratory Function using Cytochrome C Oxidase / Succinate Dehydrogenase (COX/SDH) Double-labeling Histochemistry

Mitochondrial DNA (mtDNA) defects are an important cause of disease and may underlie aging and aging-related alterations 1,2. The mitochondrial theory of aging suggests a role for mtDNA mutations, which can alter bioenergetics homeostasis and cellular function, in the aging process 3. A wealth of evidence has been compiled in support of this theory 1,4, an example being the mtDNA mutator mouse 5; however, the precise role of mtDNA damage in aging is not entirely understood 6,7.
Observing the activity of respiratory enzymes is a straightforward approach for investigating mitochondrial dysfunction. Complex IV, or cytochrome c oxidase (COX), is essential for mitochondrial function. The catalytic subunits of COX are encoded by mtDNA and are essential for assembly of the complex (Figure 1). Thus, proper synthesis and function are largely based on mtDNA integrity 2. Although other respiratory complexes could be investigated, Complexes IV and II are the most amenable to histochemical examination 8,9. Complex II, or succinate dehydrogenase (SDH), is entirely encoded by nuclear DNA (Figure 1), and its activity is typically not affected by impaired mtDNA, although an increase might indicate mitochondrial biogenesis 10-12. The impaired mtDNA observed in mitochondrial diseases, aging, and age-related diseases often leads to the presence of cells with low or absent COX activity 2,12-14. Although COX and SDH activities can be investigated individually, the sequential double-labeling method 15,16 has proved to be advantageous in locating cells with mitochondrial dysfunction 12,17-21.
Many of the optimal constitutions of the assay have been determined, such as substrate concentration, electron acceptors/donors, intermediate electron carriers, influence of pH, and reaction time 9,22,23. 3,3'-diaminobenzidine (DAB) is an effective and reliable electron donor 22. In cells with functioning COX, the brown indamine polymer product will localize in mitochondrial cristae and saturate cells 22. Those cells with dysfunctional COX will therefore not be saturated by the DAB product, allowing for the visualization of SDH activity by reduction of nitroblue tetrazolium (NBT), an electron acceptor, to a blue formazan end product 9,24. Cytochrome c and sodium succinate substrates are added to normalize endogenous levels between control and diseased/mutant tissues 9. Catalase is added as a precaution to avoid possible contaminating reactions from peroxidase activity 9,22. Phenazine methosulfate (PMS), an intermediate electron carrier, is used in conjunction with sodium azide, a respiratory chain inhibitor, to increase the formation of the final reaction products 9,25. Despite this information, some critical details affecting the result of this seemly straightforward assay, in addition to specificity controls and advances in the technique, have not yet been presented.

Visualization of Mitochondrial Respiratory Function using Cytochrome C Oxidase / Succinate Dehydrogenase COX/SDH Double-labeling Histochemistry

The cytochrome c oxidase/sodium dehydrogenase (COX/SDH) double-labeling method allows for direct visualization of mitochondrial respiratory enzyme deficiencies in fresh-frozen tissue sections. This is a straightforward histochemical technique and is useful in investigating mitochondrial diseases, aging, and aging-related disorders.

シトクロムを使用してミトコンドリア呼吸機能の可視化 Cオキシダーゼ/コハク酸脱水素酵素（COX / SDH）二重標識組織化学

Mitochondrial DNA (mtDNA) defects are an important cause of disease and may underlie aging and aging-related alterations 1,2. The mitochondrial theory of ...

Creating Defined Gaseous Environments to Study the Effects of Hypoxia on C. elegans

Oxygen is essential for all metazoans to survive, with one known exception1. Decreased O2 availability (hypoxia) can arise during states of disease, normal development or changes in environmental conditions2-5. Understanding the cellular signaling pathways that are involved in the response to hypoxia could provide new insight into treatment strategies for diverse human pathologies, from stroke to cancer. This goal has been impeded, at least in part, by technical difficulties associated with controlled hypoxic exposure in genetically amenable model organisms.
The nematode Caenorhabditis elegans is ideally suited as a model organism for the study of hypoxic response, as it is easy to culture and genetically manipulate. Moreover, it is possible to study cellular responses to specific hypoxic O2 concentrations without confounding effects since C. elegans obtain O2 (and other gasses) by diffusion, as opposed to a facilitated respiratory system6. Factors known to be involved in the response to hypoxia are conserved in C. elegans. The actual response to hypoxia depends on the specific concentration of O2 that is available. In C. elegans, exposure to moderate hypoxia elicits a transcriptional response mediated largely by hif-1, the highly-conserved hypoxia-inducible transcription factor6-9. C .elegans embryos require hif-1 to survive in 5,000-20,000 ppm O27,10. Hypoxia is a general term for "less than normal O2". Normoxia (normal O2) can also be difficult to define. We generally consider room air, which is 210,000 ppm O2 to be normoxia. However, it has been shown that C. elegans has a behavioral preference for O2 concentrations from 5-12% (50,000-120,000 ppm O2)11. In larvae and adults, hif-1 acts to prevent hypoxia-induced diapause in 5,000 ppm O212. However, hif-1 does not play a role in the response to lower concentrations of O2 (anoxia, operational definition &lt;10 ppm O2)13. In anoxia, C. elegans enters into a reversible state of suspended animation in which all microscopically observable activity ceases10. The fact that different physiological responses occur in different conditions highlights the importance of having experimental control over the hypoxic concentration of O2.
Here, we present a method for the construction and implementation of environmental chambers that produce reliable and reproducible hypoxic conditions with defined concentrations of O2. The continual flow method ensures rapid equilibration of the chamber and increases the stability of the system. Additionally, the transparency and accessibility of the chambers allow for direct visualization of animals being exposed to hypoxia. We further demonstrate an effective method of harvesting C. elegans samples rapidly after exposure to hypoxia, which is necessary to observe many of the rapidly-reversed changes that occur in hypoxia10,14. This method provides a basic foundation that can be easily modified for individual laboratory needs, including different model systems and a variety of gasses.

Creating Defined Gaseous Environments to Study the Effects of Hypoxia on C. elegans

This paper details how to use continuous-flow hypoxia chambers to generate atmospheres with defined concentrations of O2 to understand biological responses to decreased O2. This system is easy to setup and maintain, and flexible enough to suit a wide range of O2 concentrations and model systems

に低酸素の影響を検討するために定義され気体環境を作成する<em&gt; C.エレガンス</em

Oxygen is essential for all metazoans to survive, with one known exception1. Decreased O2 availability (hypoxia) can arise during states of disease, ...

An Explant Assay for Assessing Cellular Behavior of the Cranial Mesenchyme

The central nervous system is derived from the neural plate that undergoes a series of complex morphogenetic movements resulting in formation of the neural tube in a process known as neurulation. During neurulation, morphogenesis of the mesenchyme that underlies the neural plate is believed to drive neural fold elevation. The cranial mesenchyme is comprised of the paraxial mesoderm and neural crest cells. The cells of the cranial mesenchyme form a pourous meshwork composed of stellate shaped cells and intermingling extracellular matrix (ECM) strands that support the neural folds. During neurulation, the cranial mesenchyme undergoes stereotypical rearrangements resulting in its expansion and these movements are believed to provide a driving force for neural fold elevation. However, the pathways and cellular behaviors that drive cranial mesenchyme morphogenesis remain poorly studied. Interactions between the ECM and the cells of the cranial mesenchyme underly these cell behaviors. Here we describe a simple ex vivo explant assay devised to characterize the behaviors of these cells. This assay is amendable to pharmacological manipulations to dissect the signaling pathways involved and live imaging analyses to further characterize the behavior of these cells. We present a representative experiment demonstrating the utility of this assay in characterizing the migratory properties of the cranial mesenchyme on a variety of ECM components.

The cranial mesenchyme undergoes dramatic morphogenic movements that likely provides a driving force for elevation of the neural folds1,2. Here we describe a simple ex vivo explant assay to characterize the cellular behaviors of the cranial mesenchyme during neurulation. This assay has numerous applications including being amenable to pharmacological manipulations and live imaging analyses.

頭蓋間葉の細胞挙動を評価するための外植片アッセイ

The central nervous system is derived from the neural plate that undergoes a series of complex morphogenetic movements resulting in formation of the ...

C. elegans Chemotaxis Assay

Many organisms use chemotaxis to seek out food sources, avoid noxious substances, and find mates. Caenorhabditis elegans has impressive chemotaxis behavior.
 The premise behind testing the response of the worms to an odorant is to place them in an area and observe the movement evoked in response to an odorant. Even with the many available assays, optimizing worm starting location relative to both the control and test areas, while minimizing the interaction of worms with each other, while maintaining a significant sample size remains a work in progress 1-10. The method described here aims to address these issues by modifying the assay developed by Bargmann et al.1. A Petri dish is divided into four quadrants, two opposite quadrants marked "Test" and two are designated "Control". Anesthetic is placed in all test and control sites. The worms are placed in the center of the plate with a circle marked around the origin to ensure that non-motile worms will be ignored. Utilizing a four-quadrant system rather than one 2 or two 1 eliminates bias in the movement of the worms, as they are equidistant from test and control samples, regardless of which side of the origin they began. This circumvents the problem of worms being forced to travel through a cluster of other worms to respond to an odorant, which can delay worms or force them to take a more circuitous route, yielding an incorrect interpretation of their intended path. This method also shows practical advantages by having a larger sample size and allowing the researcher to run the assay unattended and score the worms once the allotted time has expired.

C. elegans Chemotaxis Assay

A method of quantitatively evaluating the chemotactic response of Caenorhabditis elegans is described. A chemotactic index (CI) was employed as a way to precisely evaluate the response of worms to certain targets, and serve as a platform of comparison between strains and compounds of interest.

C. elegansの走化性アッセイ

Many organisms use chemotaxis to seek out food sources, avoid noxious substances, and find mates.  Caenorhabditis elegans has impressive chemotaxis ...

Antibody Staining in C. Elegans Using &quot;Freeze-Cracking&quot;

To stain C. elegans with antibodies, the relatively impermeable cuticle must be bypassed by chemical or mechanical methods. "Freeze-cracking" is one method used to physically pull the cuticle from nematodes by compressing nematodes between two adherent slides, freezing them, and pulling the slides apart. Freeze-cracking provides a simple and rapid way to gain access to the tissues without chemical treatment and can be used with a variety of fixatives. However, it leads to the loss of many of the specimens and the required compression mechanically distorts the sample. Practice is required to maximize recovery of samples with good morphology. Freeze-cracking can be optimized for specific fixation conditions, recovery of samples, or low non-specific staining, but not for all parameters at once. For antibodies that require very hard fixation conditions and tolerate the chemical treatments needed to chemically permeabilize the cuticle, treatment of intact nematodes in solution may be preferred. If the antibody requires a lighter fix or if the optimum fixation conditions are unknown, freeze-cracking provides a very useful way to rapidly assay the antibody and can yield specific subcellular and cellular localization information for the antigen of interest.

Antibody Staining in C. Elegans Using &quot;Freeze-Cracking&quot;

"Freeze-cracking," a method for exposing the inner tissues of the nematode C. elegans to antibodies for protein localization, is demonstrated.

抗体染色中<em&gt; C.虫</em&gt;「フリーズクラッキング」使用

To stain C.  elegans with antibodies, the relatively impermeable cuticle must be  bypassed by chemical or mechanical methods.  "Freeze-cracking" is one ...

Large-scale Gene Knockdown in C. elegans Using dsRNA Feeding Libraries to Generate Robust Loss-of-function Phenotypes

RNA interference by feeding worms bacteria expressing dsRNAs has been a useful tool to assess gene function in C. elegans. While this strategy works well when a small number of genes are targeted for knockdown, large scale feeding screens show variable knockdown efficiencies, which limits their utility. We have deconstructed previously published RNAi knockdown protocols and found that the primary source of the reduced knockdown can be attributed to the loss of dsRNA-encoding plasmids from the bacteria fed to the animals. Based on these observations, we have developed a dsRNA feeding protocol that greatly reduces or eliminates plasmid loss to achieve efficient, high throughput knockdown. We demonstrate that this protocol will produce robust, reproducible knock down of C. elegans genes in multiple tissue types, including neurons, and will permit efficient knockdown in large scale screens. This protocol uses a commercially available dsRNA feeding library and describes all steps needed to duplicate the library and perform dsRNA screens. The protocol does not require the use of any sophisticated equipment, and can therefore be performed by any C. elegans lab.

Large-scale Gene Knockdown in C. elegans Using dsRNA Feeding Libraries to Generate Robust Loss-of-function Phenotypes

While dsRNA feeding in C. elegans is a powerful tool to assess gene function, current protocols for large scale feeding screens result in variable knockdown efficiencies. We describe an improved protocol for performing large scale RNAi feeding screens that results in highly efficacious and reproducible knockdown of gene expression.

大規模な遺伝子ノックダウン中<em&gt; C.虫</em&gt;堅牢な機能喪失表現型を生成するためのdsRNA摂食ライブラリの使用

RNA interference by feeding worms bacteria expressing dsRNAs has been a useful tool to assess gene function in C. elegans. While this strategy works well ...

An Anoxia-starvation Model for Ischemia/Reperfusion in C. elegans

Protocols for anoxia/starvation in the genetic model organism C. elegans simulate ischemia/reperfusion. Worms are separated from bacterial food and placed under anoxia for 20 hr&#160;(simulated ischemia), and subsequently moved to a normal atmosphere with food (simulated reperfusion). This experimental paradigm results in increased death and neuronal damage, and techniques are presented to assess organism viability, alterations to the morphology of touch neuron processes, as well as touch sensitivity, which represents the behavioral output of neuronal function. Finally, a method for constructing hypoxic incubators using common kitchen storage containers is described. The addition of a mass flow control unit allows for alterations to be made to the gas mixture in the custom incubators, and a circulating water bath allows for both temperature control and makes it easy to identify leaks. This method provides a low cost alternative to commercially available units.

An Anoxia-starvation Model for Ischemia/Reperfusion in C. elegans

A protocol is described that uses anoxia/starvation in C. elegans to model ischemia/reperfusion. Functional outcomes include increased mortality, visible abnormalities in GFP-labeled neuronal processes, and impaired behavioral responses that require neuronal function.

における虚血/再灌流のための無酸素飢餓モデル<em&gt; C.虫</em

Protocols for anoxia/starvation in the genetic model organism C. elegans simulate ischemia/reperfusion. Worms are separated from bacterial food and placed ...