Questa ricerca è stata condotta con finanziamenti provenienti dal Anthony M. Marchionne Foundation (70.256-23.284) e TCU IS. Gli autori ringraziano Danielle DelPriore, Amanda Morin, e Christopher Rodeheffer per i loro utili contributi per modellare questi metodi. Gli autori desiderano inoltre ringraziare Hannah Bradshaw e Randi Proffitt Leyva per la loro assistenza con le riprese di questo protocollo.

<ol>
	<li>Smith, M. A., Riby, L. M., van Eekelen, J. A. M., Foster, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glucose+enhancement+of+human+memory:+A+comprehensive+research+review+of+the+glucose+memory+facilitation+effect.">Glucose enhancement of human memory: A comprehensive research review of the glucose memory facilitation effect.</a> Neurosci Biobehav Rev. 35, 770-783 (2011).</li><li>Meikle, A., Riby, L. M., Stollery, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+glucose+ingestion+and+gluco-regulatory+control+on+cognitive+performance:+A+comparison+of+younger+and+middle+aged+adults.">The impact of glucose ingestion and gluco-regulatory control on cognitive performance: A comparison of younger and middle aged adults.</a> Hum Psychopharm Clin. 19, 523-535 (2004).</li><li>Riby, L. 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Questo manoscritto descrive una semplice procedura economico per manipolare glicemia, nonché un procedimento per determinare conseguenti variazioni accessibilità cognitiva di parole bersaglio e concetti. I metodi di cui sopra descritte possono essere applicati a una vasta gamma di aree di ricerca tra cui la psicologia sociale, psicologia cognitiva, e le scienze della nutrizione. Dato che questo metodo può essere utilizzato da persone in più aree di ricerca (alcuni che non lo fanno in genere utilizzano i dati delle i...

I ricercatori in psicologia cognitiva e delle neuroscienze hanno a lungo studiato l&#39;effetto delle fluttuazioni di glucosio nel sangue nel cervello, la cognizione e il comportamento 1. Ad esempio, i ricercatori hanno scoperto che le fluttuazioni di glucosio nel sangue prevedono differenze di memoria (verbale, digit span, lavorando, ed episodica) 1-5, attenzione 6,7, puzzle solving 8, e le prestazioni in compiti di diversi livelli di domanda cognitiva 2 , 9,10. Questa ricerca rileva che l&#39;aumento di glucosio nel sangue aumenta l&#39;attenzione e la memoria, e che questi effetti sono più forti quando si lavora su compiti cognitivamente impegnativi, e in quelli di età avanzata 1,3. Negli ultimi decenni, la ricerca sugli effetti delle fluttuazioni di glucosio nel sangue sulla psicologia umana è diventato anche un argomento di interesse nel campo della psicologia sociale 11-13. Le fluttuazioni di glucosio nel sangue sono stati collegati ai cambiamenti di umore 14, la motivazione &lt;sup&gt; 15, autocontrollo 11-13, atteggiamenti 16,17, pregiudizi 18, e alle variazioni di processo decisionale in consumatori 19, mangiare 20,21, e domini finanziari 22. Poiché questi risultati trascendono una vasta gamma di variabili dipendenti, il messaggio sul ruolo della glicemia nel processo decisionale è chiaro: di glucosio nel sangue è la forma fondamentale di energia al cervello, e svuotamenti in piombo di glucosio nel sangue a importanti cambiamenti nella cognizione e autoregolamentazione 11,13,15. Dato il ruolo importante che il glucosio nel sangue svolge in vari esiti psicologici e comportamentali, il manoscritto corrente offre un metodo semplice per la manipolazione di glucosio nel sangue che i ricercatori possono utilizzare per testare una serie di cognizioni e comportamenti. Inoltre, il manoscritto corrente fornisce un metodo per valutare l&#39;accessibilità cognitivo di parole o concetti di destinazione che può cambiare in risposta alle fluttuazioni del glucosio nel sangue usandoun compito di decisione lessicale. L&#39;efficienza dei metodi attuali li rendono ideale per la ricerca sperimentale, come manipolare efficacemente glicemia e misurano cambiamenti cognitivi con relativamente poco costosi, prodotti commercialmente disponibili. La convenienza del presente manipolazione di glucosio nel sangue rende più facile per eseguire il gran numero di partecipanti necessari per ottenere la potenza adeguata necessaria a rilevare differenze statisticamente significative nel comportamento causati da eventuali modifiche manipolati in glucosio nel sangue 23. Inoltre, il compito di decisione lessicale è utile in quanto può rivelare cambiamenti impliciti nella cognizione non facilmente accessibili attraverso misure più esplicite, come self-report esamina 24. Il compito di manipolazione di glucosio nel sangue descritto comporta un periodo di digiuno 8 ore, seguito dal consumo diretto di un calorico o una bevanda senza calorie 22. Dopo un digiuno di otto ore, i partecipanti sono assegnati in modo casuale a consumare IETil suo uno zucchero addolcita bevanda o acqua. Questo metodo consente anche per la sperimentazione di eventuali effetti unici prodotti da dolcificanti non calorici (es. Aspartame, sucralosio), che sapore dolce come dolcificanti calorici tradizionali, ma non contengono gli zuccheri necessari per aumentare i livelli di glucosio nel sangue 20,22,25 . Se interessati agli effetti delle bevande non zuccherate calorico, questo compito include un, 3 ° tra soggetti condizione facoltativa in cui i partecipanti consumano una bevanda &quot;zero calorie&quot;. L&#39;attuale metodo è stato applicato con successo in molteplici paradigmi psicologici sociali 12,18,22, e offre un modo più breve per verificare gli effetti delle variazioni di glucosio nel sangue al momento autoregolamentazione rispetto alle misure precedentemente utilizzati come test di tolleranza al glucosio 26. Gli autori forniscono anche una procedura per valutare l&#39;accessibilità cognitiva delle parole bersaglio, o concetti, utilizzando un compito di decisione lessicale 27,28. In questo task, stringhe di lettere e parole sono presentate per via telematica in formato fuoco rapido, e in genere rientrano in alcune categorie concettuali distinte (nella ricerca descritta, le categorie sono stati: parole alto contenuto di grassi alimentari, parole di alimenti a basso contenuto di grassi, non parole). I partecipanti sono invitati a identificare rapidamente (in genere tramite combinazione di tasti) se una stringa di lettere (ad esempio fhens;. Pizza) è una parola o una non-parola. La velocità di reazione in cui i partecipanti a identificare correttamente le stringhe di lettere in ogni categoria concettuale misura la relativa facilità con cui il partecipante è in grado di elaborare mentalmente le parole in quella categoria concept. Reazioni più rapide, significa che un concetto è più mentalmente accessibili e, quindi, &quot;nella mente di qualcuno&quot; 24,29,30. Di conseguenza, i compiti di decisione lessicale rendono variabili dipendenti utili, e possono anche essere utilizzati come controlli di manipolazione efficaci per misurare se un concetto recente innescata (vedi riferimento 30 per ulteriori informazioni sul priming) è infatti in seguito più Cognirelativamente accessibile di quanto sarebbe in assenza di un primo. Anche se il compito di decisione lessicale è un compito che è stato usato in psicologia sociale per decenni 29, il manoscritto corrente presenta alcune variazioni minori alla procedura, un link ad un modello di programma modificabile 27, e una lista empiricamente convalidato di parole da parte degli autori &#39; lavoro precedente che misura l&#39;accessibilità cognitiva dei sani contro i cibi non sani 20. È la speranza degli autori che l&#39;ulteriore adeguamento di queste due procedure aiuterà portare a innovazioni nei campi della ricerca psicologica sociale, del comportamento alimentare e regolazione dell&#39;energia di ricerca, e faciliterà ulteriori ricerche comparative che combina i punti di vista e la metodologia di molteplici discipline .

<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.

I partecipanti I metodi di cui sopra sono state realizzate in uno studio gestito da Hill e colleghi 20 a medie dimensioni, università privata nel sud degli Stati Uniti. La popolazione di laurea presso l&#39;Università ha fornito il campione soggetto e partecipanti hanno ricevuto credito ovviamente parziale compenso per la partecipazione allo studio. Utilizzando i metodi esatti descritti nel protocollo manoscritto ...

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.

Un metodo per la manipolazione di glucosio nel sangue ed effetti misurando i cambiamenti nella cognitiva Accessibilità di Target stimoli

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

Research

JoVE Journal

Behavior

8.9K Views.  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.

Video: 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.

Iperinsulinemico-euglicemico pinza nei topi

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

Ricerca

Medicina

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.

Iperinsulinemico-euglicemico Morsetti in Cosciente, Mouse sfrenato

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.

Insulina Microiniezione ippocampale e<em&gt; In vivo</emMicrodialisi&gt; Durante il test di memoria spaziale

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

Neuroscienze

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.

Potenziale di membrana Dye Imaging di ventromediale dell&#39;ipotalamo neuroni da topi adulti per studiare glucosio 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 ...

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.

Studiando il segnale ipotalamico insulina ad intolleranza al glucosio periferico con un sistema di infusione del farmaco continuo nel cervello del Mouse

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.

Utilizzando una combinazione di calorimetria indiretta, termografia a infrarossi e livelli di glucosio nel sangue per misurare la termogenesi del tessuto adiposo bruno nell'uomo

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

Installazione chirurgica di pinze iperglicemiche e ipoglicemizzanti nei topi

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.

Autore in primo piano: Indagine sull'omeostasi della glicemia nel cervello murino utilizzando una tecnica di clamp iperglicemica e ipoglicemica conveniente

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.

Pinza iperglicemica e pinza ipoglicemica nei topi coscienti

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

Un metodo per la manipolazione di glucosio nel sangue ed effetti misurando i cambiamenti nella cognitiva Accessibilità di Target stimoli

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Iperinsulinemico-euglicemico pinza nei topi

Iperinsulinemico-euglicemico Morsetti in Cosciente, Mouse sfrenato

Insulina Microiniezione ippocampale e

Potenziale di membrana Dye Imaging di ventromediale dell'ipotalamo neuroni da topi adulti per studiare glucosio Sensing

Studiando il segnale ipotalamico insulina ad intolleranza al glucosio periferico con un sistema di infusione del farmaco continuo nel cervello del Mouse

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

Utilizzando una combinazione di calorimetria indiretta, termografia a infrarossi e livelli di glucosio nel sangue per misurare la termogenesi del tessuto adiposo bruno nell'uomo

Pinza iperglicemica e pinza ipoglicemica nei topi coscienti

Pinza iperglicemica e pinza ipoglicemica nei topi coscienti

Pinza iperglicemica e pinza ipoglicemica nei topi coscienti