Name: a method for manipulating blood glucose and measuring resulting changes in cognitive accessibility of target stimuli
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Description: Watch this Scientific Journal Video about A Method for Manipulating Blood Glucose and Measuring Resulting Changes in Cognitive Accessibility of Target Stimuli at JoVE.com

この研究は、アンソニー・M・マルキオンネ財団（70256から23284）とTCU ISからの助成金で実施しました。著者らは、これらのメソッドを整形に対するそれらの有用な貢献のためにダニエルDelPriore、アマンダモリン、およびクリストファーRodehefferに感謝します。著者らはまた、このプロトコルの撮影とその支援のためにハンナブラッドショーとランディProffitt Leyvaのを感謝したいと思います。

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著者らは、開示することは何もありません。

この原稿は、血糖値だけでなく、ターゲット単語と概念の認知のアクセシビリティで生じる変化を測定するための手順を操作するための簡単​​で安価な手順の概要を説明します。上記に概説した方法は、社会心理学、認知心理学、栄養学などの研究分野の広い範囲に適用することができます。 （いくつかの一般的に調査データを使用しない）このメソッドは、複数の研究分野の人々によっ...

認知心理学と神経科学の研究者が長い間脳、認知、および行動1上の血糖変動の影響を検討しました。例えば、研究者は、血糖値の変動がメモリの違い（口頭、桁スパン、作業、およびエピソード）1-5、注目6,7、8を解くパズル、および認知需要2の様々なレベルのタスクの性能を予測することを見出しました、9,10。本研究では、血糖値を増やすと、注意および記憶を増強することを見つけ、そして高齢1,3のもので、認知的要求の厳しいタスクを上に作業するとき、これらの効果が最強であること。 ここ数十年では、人間の心理上の血糖変動の影響についての研究はまた、社会心理学11-13の分野に関心の話題となっています。 血糖値の変動は気分14、モチベーションの変化にリンクされています&lt;SUP&gt; 15、セルフコントロール11-13、態度16,17、偏見18、および消費者19における意思決定の変化、20,21を食べ、そして金融ドメイン22へ。これらの結果は従属変数の多種多様を超越するので、意思決定における血糖の役割についてのメッセージは明確です：血糖値は、脳へのエネルギーの重要な形態であり、そして認知の重要な変化に血糖リードで枯渇し、自己規制11,13,15。 血糖値は、さまざまな心理的、行動的アウトカムで果たす重要な役割を考えると、現在の原稿は、研究者が認知や行動の範囲をテストするために使用することができます血糖値を操作するための簡単​​な方法を提供しています。さらに、現在の原稿を使用して、血中グルコースの変動に応じて変更することができるターゲット単語または概念の認知アクセシビリティを評価するための方法を提供します字句判断タスク。彼らは効果的に血糖値を操作し、比較的安価な、市販品を使用して認知変化を測定するように、本方法の効率は、実験的研究に最適です。本血糖操作の手頃な価格は、簡単に血糖23の任意の操作の変化に起因する行動の統計的有意差を検出するために必要十分な電力を得るために必要な参加者の多くを実行することができます。それは簡単にこのような自己報告書は24の調査し、より明示的な措置を介してアクセスしていない認知の暗黙的な変化を明らかにすることができるという点で、さらに、字句判断タスクに便利です。 説明血糖操作タスクは、カロリーまたはノンカロリー飲料22の有向消費に続いて8時間の絶食期間を必要とします。 8時間絶食後、参加者はランダムにEITを消費するように割り当てられています彼女の砂糖甘味飲料や水。この方法はまた、伝統的なカロリーの甘味料のような甘い味ノンカロリー甘味料（ 例えば 、アスパルテーム、スクラロース）、によって生成される任意のユニークなエフェクトのテストは可能ですが、血糖値20,22,25を上げるために必要な糖を含有していません。非カロリー加糖飲料の効果に興味を持っている場合、このタスクは、参加者が「ゼロカロリー」飲料を消費するオプション、 第3被験者間条件を含みます。現在の方法は、複数の社会的心理的なパラダイム12,18,22に正常に適用され、耐糖能が26をテストするように以前に使用された措置よりも自己規制時の血糖値の変動の影響をテストするための短い方法を提供しています。 著者らはまた、字句決定タスク27,28を使用して、ターゲットの単語、または概念の認知のアクセシビリティを評価するための手順を提供します。このTAでSK、文字列や単語が矢継ぎ早形式でコンピュータを介して提示されており、典型的には数の異なる概念のカテゴリに分類されます（説明研究では、カテゴリは以下の通りであった。高脂肪食の言葉、低脂肪食品の単語、非単語）。参加者はすぐに文字の文字列かどうかを（一般的にキーストロークを介して）識別するよう求めている（ 例えば fhens;ピザ）単語か非単語です。参加者が正しく各概念のカテゴリに文字列を識別する反応速度は参加者が精神的にその概念のカテゴリ内の単語を処理することが可能であるとの相対的な容易さを測定します。より迅速な反応はコンセプトがより精神的にアクセス可能で、これにより、24,29,30 &quot;誰かの心に」であることを意味します。その結果、字句決定タスクは、有用な従属変数を作成し、最近でプライミングされた概念は（プライミングの詳細については30を参照する参照）確かに、その後、よりcogniであれば測定するために効果的な操作チェックとしても使用することができますそれが首相の不在にあるであろうよりもtivelyアクセス可能。字句判断タスクが何十年も29社会心理学で使用されているタスクですが、現在の原稿は&#39;プロシージャに著者から編集可能なプログラムテンプレート27、および言葉の実験的検証リストへのリンクをいくつかのマイナーなバリエーションを提示します非健康食品20対健康の認知アクセシビリティを測定する前の作業。これらの2つの手順のさらなる適応は行動とエネルギー規制研究を食べに、社会心理学的研究の分野での技術革新とリードを助けることを著者らの希望であり、さらに比較の視点を組み合わせた研究と複数の分野の方法論を容易にします。

<table><tbody><tr><td>Survey Building Software</td><td>Qualtrics</td><td>Qualtrics Research Suite</td><td>Alternative survey building softwares/applications include Survey Monkey, Google Forms, Media Lab, and Inquisit software.</td></tr><tr><td>Behavioral Task Software (for lexical decision task)</td><td>Inquisit</td><td>Inquisit 4 Lab (4.0.8.0)</td><td>Alternative behavioral task softwares/applications include Media Lab/Direct RT or programming the task into an internet browser using a programming language of your choice (such as java).</td></tr><tr><td>Batch File&amp;nbsp;</td><td>Microsoft&amp;nbsp;</td><td>Microsoft Notepad; Windows</td><td></td></tr><tr><td>Lexical Decision Task Template</td><td>Millisecond</td><td>Millisecond survey library, cited template author is linked on page</td><td>Can build a lexical decision task by hand in other behavioral task softwares.</td></tr><tr><td>Participant Scheduling and Compensation Software</td><td>SONA systems</td><td>SONA systems scheduling software</td><td>Appointments can be arranged manually, too.</td></tr><tr><td>Statistical Analysis Software</td><td>IBM</td><td>IBM SPSS Statistics Standard, 22</td><td>Alternate softwares include SAS and R.</td></tr><tr><td>Blood glucose manipulating beverege paradigm</td><td>Coca Cola&amp;nbsp;</td><td>Sprite, Sprite Zero, Sparkling water</td><td>Can use any store brand sugary beverage, non calorically sweetened beverage, and sparkling water beverage, as long as beverages are not easily discernable from each other by sight.&amp;nbsp;</td></tr><tr><td>Lancet</td><td>Assure</td><td>Assure Lanets 23 gauge</td><td>Many brands of testing lancets available both online and at local pharmacies.</td></tr><tr><td>Blood glucose testing meter</td><td>Bayer&#039;s</td><td>Breeze 2</td><td>Many brands of testing meters available both online and at local pharmacies.</td></tr><tr><td>Blood glucose testing strips</td><td>Bayer&#039;s</td><td>Breeze 2</td><td>Many brands of testing strips available both online and at local pharmacies, but they must be compatible with your chosen meter.</td></tr><tr><td>&amp;nbsp;Nitrile exam gloves (400 count)</td><td>Kirkland</td><td>Kirkland Signature Nitrile Exam Gloves</td><td>Any medical grade exam glove that provides sufficient protection from blood exposure can be used.</td></tr><tr><td>Disinfecting wipes</td><td>Lysol</td><td>Lysol Disinfecting Wipes Lemon Scent</td><td>Any wipe that can kill off any bloodborne or contact born contaminants.</td></tr></tbody></table>

a method for manipulating blood glucose and measuring resulting changes in cognitive accessibility of target stimuli

Much research in social psychology has investigated the impact of bodily energy need on cognition and decision-making. As such, blood glucose, the body's primary energy source, has been of special interest to researchers for years. Fluctuations in blood glucose have been linked to a variety of changes in cognitive and behavioral processes, such as self-control, political attitudes, and eating behavior. To help meet growing interest in the links between bodily energy need and these processes, this manuscript offers a simple methodology to experimentally manipulate blood glucose using a fasting procedure followed by administration of a sugar-sweetened, unsweetened, or artificially-sweetened beverage. This is followed by presentation of a method for measuring resulting changes in implicit cognition using a lexical decision-task. In this task, participants are asked to identify whether strings of letters are words or non-words and response latencies are recorded. Sample results from a recent publication are presented as an example of the applications for the experimental manipulation of blood glucose and the lexical decision task measures.

参加者上記の方法は、中堅企業、米国南部の私立大学でヒルと同僚20によって実行研究で実施されました。大学の学部の人口は、被験試料を提供し、参加者は、試験参加のための補償として部分的な単位時間を受け取りました。現在の原稿プロトコルで概説正確な方法を用いて、著者らは、（使用飲み物について?...

Watch this Scientific Journal Video about A Method for Manipulating Blood Glucose and Measuring Resulting Changes in Cognitive Accessibility of Target Stimuli at JoVE.com

A Method for Manipulating Blood Glucose and Measuring Resulting Changes in Cognitive Accessibility of Target Stimuli

The authors introduce a method for manipulating blood glucose and measuring resulting changes in cognitive accessibility of target words using a lexical decision task.

血糖の操作とターゲット刺激の認知アクセシビリティにおける測定結果の変化のための方法

Much research in social psychology has investigated the impact of bodily energy need on cognition and decision-making.  As such, blood glucose, the body's ...

a-method-for-manipulating-blood-glucose-measuring-resulting-changes

Universidad San Sebastian

Research

JoVE Journal

Behavior

8.9K ビュー.  Texas Christian University. The authors introduce a method for manipulating blood glucose and measuring resulting changes in cognitive accessibility of target words using a lexical decision task.

ビデオ: A Method for Manipulating Blood Glucose and Measuring Resulting Changes in Cognitive Accessibility of Target Stimuli

Hyperinsulinemic-Euglycemic Clamp in the Conscious Rat

Type 2 diabetes (T2D) is rapidly rising in prevalence. Characterized by either inadequate insulin production or the inability to utilize insulin produced, T2D results in elevated blood glucose levels. The "gold-standard" in assessing insulin sensitivity is a hyperinsulinemic-euglycemic clamp or insulin clamp. In this procedure, insulin is infused at a constant rate resulting in a drop in blood glucose. To maintain blood glucose at a constant level, exogenous glucose (D50) is infused into the venous circulation. The amount of glucose infused to maintain homeostasis is indicative of insulin sensitivity. Here, we show the basic clamp procedure in the chronically catheterized, unrestrained, conscious rat. This model allows blood to be collected with minimal stress to the animal. Following the induction of anesthesia, a midline incision is made and the left common carotid artery and right jugular vein are catheterized. Inserted catheters are flushed with heparinized saline, then exteriorized and secured. Animals are allowed to recover for 4-5 days prior to experiments, with weight gain monitored daily. Only those animals who regain weight to pre-surgery levels are used for experiments. On the day of the experiment, rats are fasted and connected to pumps containing insulin and D50. Baseline glucose is assessed from the arterial line and used a benchmark throughout the experiment (euglycemia). Following this, insulin is infused at a constant rate into the venous circulation. To match the drop in blood glucose, D50 is infused. If the rate of D50 infusion is greater than the rate of uptake, a rise in glucose will occur. Similarly, if the rate is insufficient to match whole body glucose uptake, a drop will occur. Titration of glucose continues until stable glucose readings are achieved. Glucose levels and glucose infusion rates during this stable period are recorded and reported. Results provide an index of whole body insulin sensitivity. The technique can be refined to meet specific experimental requirements. It is further enhanced by the use of radioactive tracers that can determine tissue specific insulin-stimulated glucose uptake as well as whole body glucose turnover.

The hyperinsulinemic-euglycemic clamp is the "gold standard" for the assessment of insulin action. Insulin is infused at a constant rate stimulating glucose uptake. The amount of exogenous glucose infused to counter this drop is indicative of insulin sensitivity. Here the procedure is performed on a conscious, unrestrained rat.

覚醒ラットにおける高インスリン血症、正常血糖クランプ

Type 2 diabetes (T2D) is rapidly rising in prevalence. Characterized by either inadequate insulin production or the inability to utilize insulin produced, ...

医学

Hyperinsulinemic-euglycemic Clamps in Conscious, Unrestrained Mice

Type 2 diabetes is characterized by a defect in insulin action. The hyperinsulinemic-euglycemic clamp, or insulin clamp, is widely considered the "gold standard" method for assessing insulin action in vivo. During an insulin clamp, hyperinsulinemia is achieved by a constant insulin infusion. Euglycemia is maintained via a concomitant glucose infusion at a variable rate. This variable glucose infusion rate (GIR) is determined by measuring blood glucose at brief intervals throughout the experiment and adjusting the GIR accordingly. The GIR is indicative of whole-body insulin action, as mice with enhanced insulin action require a greater GIR. The insulin clamp can incorporate administration of isotopic 2[14C]deoxyglucose to assess tissue-specific glucose uptake and [3-3H]glucose to assess the ability of insulin to suppress the rate of endogenous glucose appearance (endoRa), a marker of hepatic glucose production, and to stimulate the rate of whole-body glucose disappearance (Rd).
The miniaturization of the insulin clamp for use in genetic mouse models of metabolic disease has led to significant advances in diabetes research. Methods for performing insulin clamps vary between laboratories. It is important to note that the manner in which an insulin clamp is performed can significantly affect the results obtained. We have published a comprehensive assessment of different approaches to performing insulin clamps in conscious mice1 as well as an evaluation of the metabolic response of four commonly used inbred mouse strains using various clamp techniques2. Here we present a protocol for performing insulin clamps on conscious, unrestrained mice developed by the Vanderbilt Mouse Metabolic Phenotyping Center (MMPC; URL: www.mc.vanderbilt.edu/mmpc). This includes a description of the method for implanting catheters used during the insulin clamp. The protocol employed by the Vanderbilt MMPC utilizes a unique two-catheter system3. One catheter is inserted into the jugular vein for infusions. A second catheter is inserted into the carotid artery, which allows for blood sampling without the need to restrain or handle the mouse. This technique provides a significant advantage to the most common method for obtaining blood samples during insulin clamps which is to sample from the severed tip of the tail. Unlike this latter method, sampling from an arterial catheter is not stressful to the mouse1. We also describe methods for using isotopic tracer infusions to assess tissue-specific insulin action. We also provide guidelines for the appropriate presentation of results obtained from insulin clamps.

The hyperinsulinemic-euglycemic clamp, or insulin clamp, is the gold standard for assessing insulin action in vivo. A method for performing insulin clamps in mice is described. This includes a method for arterial catheterization that permits experiments to be performed in conscious, unrestrained mice with minimal stress.

意識、無拘束マウスにおける高インスリン血症、正常血糖クランプ

Type 2 diabetes is characterized by a defect in insulin action. The hyperinsulinemic-euglycemic clamp, or insulin clamp, is widely considered the "gold ...

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

Glucose metabolism is a useful marker for local neural activity, forming the basis of methods such as 2-deoxyglucose and functional magnetic resonance imaging. However, use of such methods in animal models requires anesthesia and hence both alters the brain state and prevents behavioral measures. An alternative method is the use of in vivo microdialysis to take continuous measurement of brain extracellular fluid concentrations of glucose, lactate, and related metabolites in awake, unrestrained animals. This technique is especially useful when combined with tasks designed to rely on specific brain regions and/or acute pharmacological manipulation; for example, hippocampal measurements during a spatial working memory task (spontaneous alternation) show a dip in extracellular glucose and rise in lactate that are suggestive of enhanced glycolysis1-3,4-5, and intrahippocampal insulin administration both improves memory and increases hippocampal glycolysis6. Substances such as insulin can be delivered to the hippocampus via the same microdialysis probe used to measure metabolites. The use of spontaneous alternation as a measure of hippocampal function is designed to avoid any confound from stressful motivators (e.g. footshock), restraint, or rewards (e.g. food), all of which can alter both task performance and metabolism; this task also provides a measure of motor activity that permits control for nonspecific effects of treatment. Combined, these methods permit direct measurement of the neurochemical and metabolic variables regulating behavior.

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

Modulation of hippocampally-dependent spatial working memory by direct intrahippocampal microinjection, accompanied and followed by in vivo microdialysis for metabolites in conscious, behaving animals.

海馬のインスリンおよびマイクロインジェクション<emインビボ&gt;</em空間記憶テスト中&gt;マイクロダイアリシス

Glucose metabolism is a useful marker for local neural activity, forming the basis of methods such as 2-deoxyglucose and functional magnetic resonance ...

神経科学

Membrane Potential Dye Imaging of Ventromedial Hypothalamus Neurons From Adult Mice to Study Glucose Sensing

Studies of neuronal activity are often performed using neurons from rodents less than 2 months of age due to the technical difficulties associated with increasing connective tissue and decreased neuronal viability that occur with age. Here, we describe a methodology for the dissociation of healthy hypothalamic neurons from adult-aged mice. The ability to study neurons from adult-aged mice allows the use of disease models that manifest at a later age and might be more developmentally accurate for certain studies. Fluorescence imaging of dissociated neurons can be used to study the activity of a population of neurons, as opposed to using electrophysiology to study a single neuron. This is particularly useful when studying a heterogeneous neuronal population in which the desired neuronal type is rare such as for hypothalamic glucose sensing neurons. We utilized membrane potential dye imaging of adult ventromedial hypothalamic neurons to study their responses to changes in extracellular glucose. Glucose sensing neurons are believed to play a role in central regulation of energy balance. The ability to study glucose sensing in adult rodents is particularly useful since the predominance of diseases related to dysfunctional energy balance (e.g.&#160;obesity) increase with age.

The activity of single neurons from adult-aged mice can be studied by dissociating neurons from specific brain regions and using fluorescent membrane potential dye imaging. By testing responses to changes in glucose, this technique can be used to study the glucose sensitivity of adult ventromedial hypothalamic neurons.

グルコース感知を勉強する成体マウス由来の膜視床下部腹内側部のニューロンの電位色素イメージング

Studies of neuronal activity are often performed using neurons from rodents less than 2 months of age due to the technical difficulties associated with ...

Studying the Hypothalamic Insulin Signal to Peripheral Glucose Intolerance with a Continuous Drug Infusion System into the Mouse Brain

Insulin regulates systematic metabolism in the hypothalamus and the peripheral insulin response. An inflammatory reaction in peripheral adipose tissues contributes to type 2 diabetes mellitus (T2DM) development and appetite regulation in the hypothalamus. Chemokine CCL5 and C-C chemokine receptor type 5 (CCR5) levels have been suggested to mediate arteriosclerosis and glucose intolerance in type 2 diabetes mellitus (T2DM). In addition, CCL5 plays a neuroendocrine role in the hypothalamus by regulating food intake and body temperature, thus, prompting us to investigate its function in hypothalamic insulin signaling and the regulation of peripheral glucose metabolism.
The micro-osmotic pump brain infusion system is a quick and precise way to manipulate CCL5 function and study its effect in the brain. It also provides a convenient alternative approach to generating a transgenic knockout animal. In this system, CCL5 signaling was blocked by intracerebroventricular (ICV) infusion of its antagonist, MetCCL5, using a micro-osmotic pump. The peripheral glucose metabolism and insulin responsiveness was detected by the Oral Glucose Tolerance Test (OGTT) and Insulin Tolerance Test (ITT). Insulin signaling activity was then analyzed by protein blot from tissue samples derived from the animals.
After 7-14 days of MetCCL5 infusion, the glucose metabolism and insulin responsiveness was impaired in mice, as seen in the results of the OGTT and ITT. The IRS-1 serine302 phosphorylation was increased and the Akt activity was reduced in mice hypothalamic neurons following CCL5 inhibition. Altogether, our data suggest that blocking CCL5 in the mouse brain increases the phosphorylation of IRS-1 S302 and interrupts hypothalamic insulin signaling, leading to a decrease in insulin function in peripheral tissues as well as the impairment of glucose metabolism.

This protocol studies the role of chemokine (C-C motif) ligand 5 (CCL5) in the hypothalamus by delivering an antagonist, MetCCL5, into the mouse brain using a micro-osmotic pump brain infusion system. This transient inhibition of CCL5 activity interrupted hypothalamic insulin signaling, leading to glucose intolerance and peripheral systemic insulin sensitivity.

マウスの脳内へ連続的な薬剤注入システムと末梢グルコース不耐症に視床下部のインスリン シグナルを勉強

Insulin regulates systematic metabolism in the hypothalamus and the peripheral insulin response. An inflammatory reaction in peripheral adipose tissues ...

Acclimation Prior to an Intraperitoneal Insulin Tolerance Test to Mitigate Stress-Induced Hyperglycemia in Conscious Mice

The insulin tolerance test is commonly used in metabolic studies to assess whole body insulin sensitivity in rodents. It is a relatively simple test that involves measurement of blood glucose levels over time following a single intraperitoneal injection of insulin. Given that it is performed in the conscious state and blood is often collected via a tail snip, it has the potential to elicit a stress response from animals due to anxiety associated with handling and blood collection. As such, a stress-induced rise in blood glucose can occur, making it difficult to detect and interpret the primary endpoint measure, namely an insulin-mediated reduction in blood glucose. This has been seen in many mouse strains, and is quite common in diabetic db/db mice, where glucose levels can increase, rather than decrease, after insulin administration. Here, we describe a method of acclimating mice to handling, injections and blood sampling prior to performing the insulin tolerance test. We find that this lowers stress-induced hyperglycemia and results in data that more accurately reflects whole body insulin sensitivity.

Stress-induced elevations in glucose levels can confound interpretation of data derived from a conscious intraperitoneal insulin tolerance test in mice. In this article, we describe a method to acclimate the mice to handling, injections and blood sampling prior to performing the insulin tolerance test in order to limit stress-induced hyperglycemia.

The insulin tolerance test is commonly used in metabolic studies to assess whole body insulin sensitivity in rodents. It is a relatively simple test that ...

Using a Combination of Indirect Calorimetry, Infrared Thermography, and Blood Glucose Levels to Measure Brown Adipose Tissue Thermogenesis in Humans

In mammals, brown adipose tissue (BAT) is activated rapidly in response to cold in order to maintain body temperature. Although BAT has been studied greatly in small animals, it is difficult to measure the activity of BAT in humans. Therefore, little is known about the heat-generating capacity and physiological significance of BAT in humans, including the degree to which components of the diet can activate BAT. This is due to the limitations in the currently most used method to assess the activation of BAT-radiolabeled glucose (fluorodeoxyglucose or 18FDG) measured by positron emission tomography-computerized tomography (PET-CT).
This method is usually performed in fasted subjects, as feeding induces glucose uptake by the muscles, which can mask the glucose uptake into the BAT. This paper describes a detailed protocol for quantifying total-body human energy expenditure and substrate utilization from BAT thermogenesis by combining indirect calorimetry, infrared thermography, and blood glucose monitoring in carbohydrate-loaded adult males. To characterize the physiological significance of BAT, measures of the impact of BAT activity on human health are critical. We demonstrate a protocol to achieve this by combining carbohydrate loading and indirect calorimetry with measurements of supraclavicular changes in temperature. This novel approach will help to understand the physiology and pharmacology of BAT thermogenesis in humans.

Here, we present a protocol to quantify the physiological significance of the impact of brown adipose tissue (BAT) activity on human metabolism. This is achieved by combining carbohydrate loading and indirect calorimetry with measurements of supraclavicular changes in temperature. This novel approach can help develop a pharmacological target for BAT thermogenesis in humans.

間接熱量測定、赤外線サーモグラフィ、および血糖値の組み合わせを使用して、ヒトの褐色脂肪組織の熱発生を測定します

In mammals, brown adipose tissue (BAT) is activated rapidly in response to cold in order to maintain body temperature. Although BAT has been studied ...

マウスにおける高血糖および低血糖クランプの外科的設置

Hyperglycemic Clamp and Hypoglycemic Clamp in Conscious Mice

Diabetes mellitus (DM) is caused by insufficient insulin release from the pancreatic &#946;-cells (Type1 DM) and insulin sensitivity in muscles, liver, and adipose tissues (Type2 DM). Insulin injection treats DM patients but leads to hypoglycemia as a side effect. Cortisol and catecholamines are released to activate glucose production from the liver to recover hypoglycemia, called counter-regulatory responses (CRR). In DM research using rodent models, glucose tolerance tests and 2-deoxy-glucose injection are used to measure insulin release and CRR, respectively. However, blood glucose concentrations change persistently during experiments, causing difficulties in assessing net insulin release and CRR. This article describes a method in which blood glucose is kept at 250 mg/dL or 50 mg/dL in conscious mice to compare the release of insulin and CRR hormones, respectively.
Polyethylene tubing is implanted in the mice's carotid artery and jugular vein, and the mice are allowed to recover from the surgery. The jugular vein tubing is connected to a Hamilton syringe with a syringe pump to enable insulin or glucose infusion at a constant and variable rate. The carotid artery tubing is for blood collection. For the hyperglycemic clamp, 30% glucose is infused into the vein, and blood glucose levels are measured from the arterial blood every 5 min or 10 min. The infusion rate of 30% glucose is increased until the blood glucose level becomes 250 mg/dL. Blood is collected to measure insulin concentrations. For hypoglycemic clamp, 10 mU/kg/min insulin is infused together with 30% glucose, whose infusion rate is variable to maintain 50 mg/dL of blood glucose level. Blood is collected to measure counter-regulatory hormones when both glucose infusion and blood glucose reach a steady state. Both hyperglycemic and hypoglycemic clamps have the same surgical procedure and experimental setups. Thus, this method is useful for researchers of systemic glucose metabolism.

著者スポットライト:費用対効果の高い高血糖および低血糖クランプ技術を使用したマウス脳の血糖恒常性の調査

A hyperglycemic clamp is used for measuring insulin release with a maintained higher blood glucose concentration. A hypoglycemic clamp is for measuring glucose production induced by counter-regulatory responses. Both methods use the same surgical procedure. Here, we present a clamp technique to assess systemic glucose metabolism.

意識のあるマウスにおける高血糖クランプと低血糖クランプ

Diabetes mellitus (DM) is caused by insufficient insulin release from the pancreatic &#946;-cells (Type1 DM) and insulin sensitivity in muscles, liver, ...

血糖の操作とターゲット刺激の認知アクセシビリティにおける測定結果の変化のための方法

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覚醒ラットにおける高インスリン血症、正常血糖クランプ

意識、無拘束マウスにおける高インスリン血症、正常血糖クランプ

海馬のインスリンおよびマイクロインジェクション

グルコース感知を勉強する成体マウス由来の膜視床下部腹内側部のニューロンの電位色素イメージング

マウスの脳内へ連続的な薬剤注入システムと末梢グルコース不耐症に視床下部のインスリンシグナルを勉強

Acclimation Prior to an Intraperitoneal Insulin Tolerance Test to Mitigate Stress-Induced Hyperglycemia in Conscious Mice

間接熱量測定、赤外線サーモグラフィ、および血糖値の組み合わせを使用して、ヒトの褐色脂肪組織の熱発生を測定します

意識のあるマウスにおける高血糖クランプと低血糖クランプ

意識のあるマウスにおける高血糖クランプと低血糖クランプ

意識のあるマウスにおける高血糖クランプと低血糖クランプ

血糖の操作とターゲット刺激の認知アクセシビリティにおける測定結果の変化のための方法

要約

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覚醒ラットにおける高インスリン血症、正常血糖クランプ

意識、無拘束マウスにおける高インスリン血症、正常血糖クランプ

海馬のインスリンおよびマイクロインジェクション

グルコース感知を勉強する成体マウス由来の膜視床下部腹内側部のニューロンの電位色素イメージング

マウスの脳内へ連続的な薬剤注入システムと末梢グルコース不耐症に視床下部のインスリン シグナルを勉強

Acclimation Prior to an Intraperitoneal Insulin Tolerance Test to Mitigate Stress-Induced Hyperglycemia in Conscious Mice

間接熱量測定、赤外線サーモグラフィ、および血糖値の組み合わせを使用して、ヒトの褐色脂肪組織の熱発生を測定します

意識のあるマウスにおける高血糖クランプと低血糖クランプ

意識のあるマウスにおける高血糖クランプと低血糖クランプ

意識のあるマウスにおける高血糖クランプと低血糖クランプ

マウスの脳内へ連続的な薬剤注入システムと末梢グルコース不耐症に視床下部のインスリンシグナルを勉強