Name: measuring glutathione-induced feeding response in hydra
Uploaded: 2014-11-16T15:00:00.000+00:00
Duration: 8 min 10 s
Description: Watch this Scientific Journal Video about Measuring Glutathione-induced Feeding Response in Hydra at JoVE.com

Authors are thankful to K. P. Madhu, Nita Beliappa and staff of the Media Centre of Indian Institute of Science Education and Research, Pune for their help in the video production. The work was supported by funding under the Centre of Excellence program of Department of Biotechnology, Government of India to SG and postdoctoral fellowship by Department of Science and Technology, Government of India to RK.

摂食応答1.ハイドラ文化と測定<ol><li>アルテミア毎日それらを供給し、中に含まれる培地（1mMのTris-HCl緩衝液、pHは7.6、1のNaCl、1mMのCaCl 2、0.1mMのKClと0.1mMのMgSO 4）し、それらを保持することにより、培養中ヒドラポリープを維持12時間明、12時間暗サイクルの下で18℃、ガラスボウル12前述したように。 </li><li>摂食応答を測定するために、一つは24ウェルプレートの1ウェルに5~6触手を有するヒドラポリープ成熟移す。それを傾けることによってウェルから残留培地を除去した後、すぐに新鮮な培地500μlを加える。 </li><li>ヒドラ培地中の9100μMのグルタチオン溶液を調製する。グルタチオン溶液は酸化されやすいので、常に各実験のために新たに調製したグルタチオン溶液を使用する。 </li><li>画像記録のための条項を持つ顕微鏡のイメージングプラットフォームにプレートを転送します。行動oは、そのような暗い背景を使用してくださいfはヒドラポリープは明らかに対照的な背景に画像化することができる。 </li><li>部屋は観察し、変動強度、空気流及び騒音の光からヒドラフリーの挙動を撮像するために使用される。でも、グルタチオンが存在しない場合に - このような妨害はまたヒドラポリープは触手の収縮を示すことが原因となることがあります。 </li><li>ポリープは5分間リラックスすることができます。 </li><li>ポリープは行動が明確に画像化することができることをよくかかるの中心領域に沿って配置されていることを確認してください。ポリープはウェルの端にある場合には、ピペットを用いて培地をフラッシュすることによって、中心に持ってきて、もう一度それがリラックスすることができます。 </li><li>リラックスした状態でヒドラのイメージをキャプチャします。これは、ゼロ時点で観察されます。 </li><li>迅速にウェル内に、3μMの最終濃度に到達するために9μMグルタチオン溶液を加える。実験の目的及びヒドラで示される反応に応じて、グルタチオンおよびチュウの異なる濃度範囲を試験必要とされる適切な濃度のSeを示す。 </li><li>すぐに4-5分ごとに15〜30秒後にグルタチオン溶液とキャプチャ画像を追加した後、タイマーを起動します。タイムラプスイメージング中倍率設定を変更しないでください。 </li><li>穏やかにウェル内の動物の位置を最小限に顕微鏡の視野内に乱されることを均一な流れとグルタチオン溶液を加える。ポリープは、グルタチオン溶液を添加した後に広範囲に移動する場合は、画像キャプチャのための視野内にポリープをもたらすために非常に穏やかにプレートを移動させる。 </li><li>対照実験では、同一の他の全てのパラメータを保持しながら媒体グルタチオンを欠く使用。 </li><li>午後1時前に摂食応答の程度に概日リズムの可能性のある影響を避けるために - その日の前半に上記の実験の手順をすべて実行してください。 </li><li> GNU画像処理プログラム（GIMP）を使用して撮影した画像のそれぞれを開きます。 </lI&gt; <li>触手とhypostomeのそれぞれの先端部間の距離を決定するために測定&gt;メニュー&gt;ツールから利用できる「測定」ツールを使用します。口の開口部は、画像のいずれにおいても観察されている場合は、開いた口の中心と触手の先端部間の距離を決定する。触手の広がりとしてこの距離を参照してください。 </li><li>グルタチオンの暴露の前後に各ポリープの平均触手スプレッドを計算します。その後の時点の各々でそのまでゼロ時点での平均触手スプレッドの比を計算する。この比は、相対的な触手の広がりと呼ぶことにする。 </li><li>少なくとも20ポリープのための測定を繰り返します。 </li></ol><img alt="式（1）" fo:content-width="3.5in" src="/files/ftp_upload/52178/52178eq1.jpg" width="400" /> <img alt="式2" fo:content-width="4in" src="/files/ftp_upload/52178/52178eq2.jpg" width="450" /> 2.メソッドバリデーション使用飢餓モデル<ol><li>飢餓のために、別のガラスボウルに少数のヒドラポリープを転送し、5日間のためにそれらを餌を与えないでください。同じようなサイズのボウルにアルテミアを毎日、数ポリープの対照群を養う。実験の両方からメディアを変更して培地中の真菌の増殖を避けるために、毎日ボウル。 </li><li>実験の日に、1時間アルテミアとヒドラの対照群を供給し、メディアからすべての食べ残しや死者アルテミアを除去した後、その後の実験のためにこれらのヒドラを使用しています。 </li><li>飢餓および制御ヒドラポリープのそれぞれの測定値を交互による観察時に任意のバイアスを避けるために、以前のステップ1に記載の方法により対照群と比較して、ヒドラ飢餓ヒドラの摂食応答を測定する。 </li></ol>

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The authors declare no competing financial interests.

ヒドラに摂食行動することは後生動物の中で最も祖先の化学感覚システムのいずれかを表します。刺胞アシスト獲物捕獲後にリリース甲殻流体中のGSHの存在はずっと前に1を検出されたが、GSHRタンパク質も推定されるコード遺伝子/ sのどちらも、ヒドラから現在までに特徴付けられている。いくつかの試みがヒドラ8、14、15結合タンパク質GSHを特徴付けるために行われている?...

Hydra is the most primitive organism possessing a nervous system and chemosensation for detecting reduced glutathione (GSH) for capturing the prey1. It feeds on a variety of animals such as nematode, crustacea, insect larvae, tadpoles and newly hatched fish1. The movement of these prey organisms causes mechanosensory discharge of the stinging capsules called nematocysts from hydra, which are inserted into the prey2. GSH present in the fluid released from the injured prey subsequently activates the feeding response in hydra which includes curling of the tentacles to bring the prey towards the mouth, opening of the mouth, and consequent engulfing of the prey. Multiple molecules, such as dopamine3, glutamate4, GABA, glycine5, NMDA receptors6, and allatotropin7, have been shown to be involved in the feeding response in hydra. It has also been shown that the chemosensory response induced by GSH is modulated by the feeding status of the animal such that starved hydra exhibited enhanced feeding response1. Such an increase in the GSH sensitivity is biologically relevant since under starvation hydra need to find its prey at higher sensitivity. 
Although the feeding response induced by GSH can be clearly observed under microscope, the methods typically used for measuring the feeding response observations are non-quantitative. In most of the cases, the time during which the mouth of the hydra remains open was taken as a measure of the feeding response8,9; whereas in another case, quantitation was based on the number of hydra out of a population showing the feeding response10. However, observing the mouth opening time of the hydra polyps is cumbersome and subject to variation induced by uncontrollable parameters such as the direction of the mouth orientation during observations. Similarly, since the feeding response is a quantitative parameter, population-based approaches are subject to variations/errors caused by the opinion or observational bias of the individual observer. To circumvent these issues we have developed a method for the relative quantification of the feeding response in hydra (Hydra vulgaris Ind-Pune11) based on the distance of the apical end of the tentacle from the mouth of the hydra polyp.

<table><tbody><tr><td>Cooled Incubator</td><td>Panasonic&amp;nbsp;</td><td>MIR-254-PE</td><td></td></tr><tr><td>Microscope</td><td>Leica</td><td>S8AP0&amp;nbsp;</td><td></td></tr><tr><td>Camera for the microscope</td><td>Leica&amp;nbsp;</td><td>EC3</td><td></td></tr><tr><td>Reduced glutathione</td><td>Sigma</td><td>G4251</td><td>Stored at 4 &amp;deg;C. Bring the bottle to room temperature before opening to avoid oxidation</td></tr><tr><td>Image editing program</td><td>GIMP</td><td>Version 2.8</td><td></td></tr></tbody></table>

measuring glutathione-induced feeding response in hydra

Hydra is among the most primitive organisms possessing a nervous system and chemosensation for detecting reduced glutathione (GSH) for capturing the prey. The movement of prey organisms causes mechanosensory discharge of the stinging cells called nematocysts from hydra, which are inserted into the prey. The feeding response in hydra, which includes curling of the tentacles to bring the prey towards the mouth, opening of the mouth and consequent engulfing of the prey, is triggered by GSH present in the fluid released from the injured prey. To be able to identify the molecular mechanism of the feeding response in hydra which is unknown to date, it is necessary to establish an assay to measure the feeding response. Here, we describe a simple method for the quantitation of the feeding response in which the distance between the apical end of the tentacle and mouth of hydra is measured and the ratio of such distance before and after the addition of GSH is determined. The ratio, called the relative tentacle spread, was found to give a measure of the feeding response. This assay was validated using a starvation model in which starved hydra show an enhanced feeding response in comparison with daily fed hydra.

グルタチオンは、ヒドラが獲物を飲み込むの目的のために、口の方に触手のカールを呈するようになります。触手のようなカールが近いhypostomeに触手の先端両端をもたらします。これは触手の広がりの減少、または触手とhypostome（ 図1）の先端部間の直線距離が生じる。相対触手広がり、または平均触手の比率は、前に広がり、グルタチオンを添加した後、時間の経過とともに減...

Watch this Scientific Journal Video about Measuring Glutathione-induced Feeding Response in Hydra at JoVE.com

Measuring Glutathione-induced Feeding Response in Hydra

Here we describe a simple assay for the quantification of the feeding response in hydra induced by the reduced form of glutathione. This assay relies on measuring the distance between the apical end of the tentacle and mouth of hydra.

ハイドラ内グルタチオンによって誘発される摂食応答を測定する

Hydra is among the most primitive organisms possessing a nervous system and chemosensation for detecting reduced glutathione (GSH) for capturing the prey. ...

measuring-glutathione-induced-feeding-response-in-hydra

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

11.3K ビュー.  India Institute of Science Education and Research, Pune. Here we describe a simple assay for the quantification of the feeding response in hydra induced by the reduced form of glutathione. This assay relies on measuring the distance between the apical end of the tentacle and mouth of hydra.

ビデオ: Measuring Glutathione-induced Feeding Response in Hydra

Imaging Approaches to Assessments of Toxicological Oxidative Stress Using Genetically-encoded Fluorogenic Sensors

While oxidative stress is a commonly cited toxicological mechanism, conventional methods to study it suffer from a number of shortcomings, including destruction of the sample, introduction of potential artifacts, and a lack of specificity for the reactive species involved. Thus, there is a current need in the field of toxicology for non-destructive, sensitive, and specific methods that can be used to observe and quantify intracellular redox perturbations, more commonly referred to as oxidative stress. Here, we present a method for the use of two genetically-encoded fluorogenic sensors, roGFP2 and HyPer, to be used in live-cell imaging studies to observe xenobiotic-induced oxidative responses. roGFP2 equilibrates with the glutathione redox potential (EGSH), while HyPer directly detects hydrogen peroxide (H2O2). Both sensors can be expressed into various cell types via transfection or transduction, and can be targeted to specific cellular compartments. Most importantly, live-cell microscopy using these sensors offers high spatial and temporal resolution that is not possible using conventional methods. Changes in the fluorescence intensity monitored at 510 nm serves as the readout for both genetically-encoded fluorogenic sensors when sequentially excited by 404 nm and 488 nm light. This property makes both sensors ratiometric, eliminating common microscopy artifacts and correcting for differences in sensor expression between cells. This methodology can be applied across a variety of fluorometric platforms capable of exciting and collecting emissions at the prescribed wavelengths, making it suitable for use with confocal imaging systems, conventional wide-field microscopy, and plate readers. Both genetically-encoded fluorogenic sensors have been used in a variety of cell types and toxicological studies to monitor cellular EGSH and H2O2 generation in real-time. Outlined here is a standardized method that is widely adaptable across cell types and fluorometric platforms for the application of roGFP2 and HyPer in live-cell toxicological assessments of oxidative stress.

This manuscript describes the use of genetically-encoded fluorogenic reporters in an application of live-cell imaging for the examination of xenobiotic-induced oxidative stress. This experimental approach offers unparalleled spatiotemporal resolution, sensitivity, and specificity while avoiding many of the shortcomings of conventional methods used for the detection of toxicological oxidative stress.

遺伝子にコードされた蛍光センサーを用いた毒性酸化ストレスの評価へのアプローチをイメージング

While oxidative stress is a commonly cited toxicological mechanism, conventional methods to study it suffer from a number of shortcomings, including ...

がん研究

Combining Quantitative Food-intake Assays and Forcibly Activating Neurons to Study Appetite in Drosophila

Food consumption is under the tight control of the brain, which integrates the physiological status, palatability, and nutritional contents of the food, and issues commands to start or stop feeding. Deciphering the processes underlying the decision-making of timely and moderate feeding carries major implications in our understanding of physiological and psychological disorders related to feeding control. Simple, quantitative, and robust methods are required to measure the food ingestion of animals after experimental manipulation, such as forcibly increasing the activities of certain target neurons. Here, we introduced dye-labeling-based feeding assays to facilitate the neurogenetic study of feeding control in adult fruit flies. We review available feeding assays, and then describe our methods step-by-step from setup to analysis, which combine thermogenetic and optogenetic manipulation of neurons controlling feeding motivation with dye-labeled food intake assay. We also discuss the advantages and limitations of our methods, compared with other feeding assays, to help readers choose an appropriate assay.

Combining Quantitative Food-intake Assays and Forcibly Activating Neurons to Study Appetite in Drosophila

Quantitative food-intake assays with dyed food provide a robust and high-throughput means to evaluate feeding motivation. Combining the food consumption assay with thermogenetic and optogenetic screens is a powerful approach to investigate the neural circuits underlying appetite in adult Drosophila melanogaster.

定量的食物摂取の試金を組み合わせると強制的にショウジョウバエの食欲を勉強するニューロンを活性化

Food consumption is under the tight control of the brain, which integrates the physiological status, palatability, and nutritional contents of the food, ...

行動学

Measurement of Glutamate Uptake using Radiolabeled L-[3H]-Glutamate in Acute Transverse Slices Obtained from Rodent Resected Hippocampus

Glutamate removal from the extracellular space by high-affinity Na+-dependent transporters is essential to ensure that the brain&#39;s intrinsic connectivity mechanisms work properly and homeostasis is maintained. The hippocampus is a unique brain structure that manages higher cognitive functions, and is the subject of several studies regarding neurologic diseases. The investigation of physiological and pathological mechanisms in rodent models can benefit from acute hippocampal slice (AHS) preparations. AHS has the advantage of providing reliable information on cell function since the cytoarchitecture and synaptic circuits are preserved. Although AHS preparations are commonly used in neurochemistry laboratories, it is possible to find some methodological differences in the literature. Considering that distinctive slice preparation protocols might change the hippocampal regions analyzed, this current protocol proposes a standard technique for obtaining transverse AHS from resected hippocampus. This simple-to-perform protocol may be used in mice and rats&#39; experimental models and allow several ex vivo approaches investigating neurochemical dynamics (in dorsal, intermediate and ventral hippocampus) in different backgrounds (e.g., transgenic manipulations) or after in vivo manipulations (e.g., pharmacological treatments or suitable rodent models to study clinical disorders). After dissecting the hippocampus from the rodent brain, transverse slices along the septo-temporal axis (300 &#181;m thick) were obtained. These AHS contain distinct parts of the hippocampus and were subjected to an individual neurochemical investigation (as an example: neurotransmitter transporters using their respective substrates). As the hippocampus presents a high density of excitatory synapses, and glutamate is the most important neurotransmitter in the brain, the glutamatergic system is an interesting target for in vivo observed phenomena. Thus, the current protocol provides detailed steps to explore glutamate uptake in ex vivo AHS using L-[3H]-Glutamate. Using this protocol to investigate hippocampal function may help to better understand the influence of glutamate metabolism on mechanisms of neuroprotection or neurotoxicity.

Measurement of Glutamate Uptake using Radiolabeled L-[3H]-Glutamate in Acute Transverse Slices Obtained from Rodent Resected Hippocampus

This article describes a reliable and simple way to obtain ex vivo acute hippocampal transverse slices from mice and rats using a tissue chopper. Slices obtained from resected hippocampi can be submitted for functional glutamate uptake analysis to investigate glutamatergic system homeostasis.

Glutamate removal from the extracellular space by high-affinity Na+-dependent transporters is essential to ensure that the brain&#39;s intrinsic ...

生化学

線虫の食物摂取量と寿命を測定するためのハイスループットアッセイ

Quantifying Food Intake in Caenorhabditis elegans by Measuring Bacterial Clearance

Feeding is an essential biological process for an organism&#39;s growth, reproduction, and survival. This assay aims to measure the food intake of Caenorhabditis elegans&#160;(C. elegans), an important parameter when studying the genetics of aging or metabolism. In most species, feeding is determined by measuring the difference between the amount of food provided and the amount left after a given time interval. The method presented here uses the same strategy to determine the feeding of C. elegans. It measures the amount of bacteria, the food source of C. elegans, cleared within 72 h. This method uses 96-well microtiter plates and has allowed the screening of hundreds of drugs for their ability to modulate food intake at a speed and depth not possible in other animal models. The strength of this assay is that it allows to measure feeding and lifespan simultaneously and directly measures the disappearance of food and, thus, is based on the same principles used for other organisms, facilitating species-to-species comparison.

著者スポットライト:脂肪毒性とHFpEFの関係を探る 

This protocol describes an assay to quantify Caenorhabditis elegans feeding rate based on measuring the clearance of bacteria in liquid culture.

細菌クリアランスの測定による 線虫 の食物摂取量の定量化

Feeding is an essential biological process for an organism&#39;s growth, reproduction, and survival. This assay aims to measure the food intake of ...

GSH/GSSG 測定 – 抗酸化物質研究のための精製 OPA アッセイ

Rapid Quantification of Oxidized and Reduced Forms of Glutathione Using Ortho -phthalaldehyde in Cultured Mammalian Cells In Vitro

Glutathione has long been considered a key biomarker for determining the antioxidant response of the cell. Hence, it is a primary marker for reactive oxygen species studies. The method utilizes Ortho-phthalaldehyde (OPA) to quantify the cellular concentration of glutathione(s). OPA conjugates with reduced glutathione (GSH) via sulfhydryl binding to subsequently form an isoindole, resulting in a highly fluorescent conjugate. To attain an accurate result of both oxidized glutathione (GSSG) and GSH, a combination of masking agents and reducing agents, which have been implemented in this protocol, are required. Treatments may also impact cellular viability. Hence, normalization via protein assay is presented in this multiparametric assay. The assay demonstrates a pseudo-linear detection range of 0.234 - 30&#181;M (R2=0.9932&#177;0.007 (N=12)) specific to GSH. The proposed assay also allows for the determination of oxidized glutathione with the addition of the masking agent N-ethylmaleimide to bind reduced glutathione, and the reducing agent tris(2-carboxyethyl) phosphine is introduced to cleave the disulfide bond in GSSG to produce two molecules of GSH. The assay is used in combination with a validated bicinchoninic acid assay for protein quantification and an adenylate kinase assay for cytotoxicity assessment.

著者スポットライト:酸化ストレス研究のためのチオール定量とバイオマーカー検出の革新 

Quantification of both oxidized and reduced forms of glutathione (GSSG and GSH, respectively) has been achieved through the use of Ortho-phthalaldehyde (OPA). OPA becomes highly fluorescent once conjugated to GSH but is unable to conjugate GSSG until reduced. Here, we describe a multiparametric assay to quantify both using protein quantification for normalization.

培養哺乳類細胞における オルト -フタルアルデヒドを用いたグルタチオンの酸化型および還元型グルタチオンの迅速な定量(英語)

Glutathione has long been considered a key biomarker for determining the antioxidant response of the cell. Hence, it is a primary marker for reactive ...

Generation and Long-term Maintenance of Nerve-free Hydra

The interstitial cell lineage of Hydra includes multipotent stem cells, and their derivatives: gland cells, nematocytes, germ cells, and nerve cells. The interstitial cells can be eliminated through two consecutive treatments with colchicine, a plant-derived toxin that kills dividing cells, thus erasing the potential for renewal of the differentiated cells that are derived from the interstitial stem cells. This allows for the generation of Hydra that lack nerve cells. A nerve-free polyp cannot open its mouth to feed, egest, or regulate osmotic pressure. Such animals, however, can survive and be cultured indefinitely in the laboratory if regularly force-fed and burped. The lack of nerve cells allows for studies of the role of the nervous system in regulating animal behavior and regeneration. Previously published protocols for nerve-free Hydra maintenance involve outdated techniques such as mouth-pipetting with hand-pulled micropipette tips to feed and clean the Hydra. Here, an improved protocol for maintenance of nerve-free Hydra is introduced. Fine-tipped forceps are used to force open the mouth and insert freshly killed Artemia. Following force-feeding, the body cavity of the animal is flushed with fresh medium using a syringe and hypodermic needle to remove undigested material, referred to here as "burping". This new method of force-feeding and burping nerve-free Hydra through the use of forceps and syringes eliminates the need for mouth-pipetting using hand-pulled micropipette tips. It thus makes the process safer and significantly more time efficient. To ensure that the nerve cells in the hypostome have been eliminated, immunohistochemistry using anti-tyrosine-tubulin is conducted.

Generation and Long-term Maintenance of Nerve-free Hydra

Through a double treatment with colchicine, a plant-derived toxin that kills dividing cells, nerve-free Hydra vulgaris can be generated. These Hydra cannot feed or egest on their own. This paper describes an improved method for long-term maintenance of nerve free Hydra vulgaris in the laboratory.

神経フリーの生成と長期維持<em&gt;ヒドラ</em

The interstitial cell lineage of Hydra includes multipotent stem cells, and their derivatives: gland cells, nematocytes, germ cells, and nerve cells. The ...

ハイドラ内グルタチオンによって誘発される摂食応答を測定する

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この動画の章

遺伝子にコードされた蛍光センサーを用いた毒性酸化ストレスの評価へのアプローチをイメージング

定量的食物摂取の試金を組み合わせると強制的にショウジョウバエの食欲を勉強するニューロンを活性化

Measurement of Glutamate Uptake using Radiolabeled L-[³H]-Glutamate in Acute Transverse Slices Obtained from Rodent Resected Hippocampus

細菌クリアランスの測定による線虫の食物摂取量の定量化

細菌クリアランスの測定による線虫の食物摂取量の定量化

細菌クリアランスの測定による線虫の食物摂取量の定量化

培養哺乳類細胞におけるオルト -フタルアルデヒドを用いたグルタチオンの酸化型および還元型グルタチオンの迅速な定量(英語)

培養哺乳類細胞におけるオルト -フタルアルデヒドを用いたグルタチオンの酸化型および還元型グルタチオンの迅速な定量(英語)

培養哺乳類細胞におけるオルト -フタルアルデヒドを用いたグルタチオンの酸化型および還元型グルタチオンの迅速な定量(英語)

神経フリーの生成と長期維持

ハイドラ内グルタチオンによって誘発される摂食応答を測定する

要約

さらに動画を探す

この動画の章

遺伝子にコードされた蛍光センサーを用いた毒性酸化ストレスの評価へのアプローチをイメージング

定量的食物摂取の試金を組み合わせると強制的にショウジョウバエの食欲を勉強するニューロンを活性化

Measurement of Glutamate Uptake using Radiolabeled L-[3H]-Glutamate in Acute Transverse Slices Obtained from Rodent Resected Hippocampus

細菌クリアランスの測定による 線虫 の食物摂取量の定量化

細菌クリアランスの測定による 線虫 の食物摂取量の定量化

細菌クリアランスの測定による 線虫 の食物摂取量の定量化

培養哺乳類細胞における オルト -フタルアルデヒドを用いたグルタチオンの酸化型および還元型グルタチオンの迅速な定量(英語)

培養哺乳類細胞における オルト -フタルアルデヒドを用いたグルタチオンの酸化型および還元型グルタチオンの迅速な定量(英語)

培養哺乳類細胞における オルト -フタルアルデヒドを用いたグルタチオンの酸化型および還元型グルタチオンの迅速な定量(英語)

神経フリーの生成と長期維持

Measurement of Glutamate Uptake using Radiolabeled L-[³H]-Glutamate in Acute Transverse Slices Obtained from Rodent Resected Hippocampus

細菌クリアランスの測定による線虫の食物摂取量の定量化

細菌クリアランスの測定による線虫の食物摂取量の定量化

細菌クリアランスの測定による線虫の食物摂取量の定量化

培養哺乳類細胞におけるオルト -フタルアルデヒドを用いたグルタチオンの酸化型および還元型グルタチオンの迅速な定量(英語)

培養哺乳類細胞におけるオルト -フタルアルデヒドを用いたグルタチオンの酸化型および還元型グルタチオンの迅速な定量(英語)

培養哺乳類細胞におけるオルト -フタルアルデヒドを用いたグルタチオンの酸化型および還元型グルタチオンの迅速な定量(英語)