Esta pesquisa foi realizada com subvenções do M. Marchionne Fundação Anthony (70.256-23.284) e TCU IS. Os autores agradecem Danielle DelPriore, Amanda Morin, e Christopher Rodeheffer por suas contribuições úteis para moldar esses métodos. Os autores também gostaria de agradecer a Hannah Bradshaw e Randi Proffitt Leyva por sua assistência com a filmar este protocolo.

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</ol>

Os autores não têm nada para revelar.

Este manuscrito descreve um procedimento simples, de baixo custo para a manipulação de glucose no sangue, bem como um procedimento para a medição das alterações resultantes na acessibilidade cognitiva de palavras e conceitos alvo. Os métodos descritos acima podem ser aplicados a uma ampla gama de áreas de investigação incluindo psicologia social, psicologia cognitiva e ciências da nutrição. Dado que este método pode ser usado por pessoas em várias áreas de pesquisa (alguns que não costumam usar dados de...

Pesquisadores em psicologia cognitiva e neurociência há muito que estudam o impacto das flutuações de glicose no sangue no cérebro, cognição e comportamento 1. Por exemplo, os pesquisadores descobriram que as flutuações nos níveis de glicose no sangue prever diferenças na memória (verbal, extensão de dígitos, trabalhar e episódica) 1-5, a atenção 6,7, resolver quebra-cabeças 8, e o desempenho em tarefas de diferentes níveis de demanda cognitiva 2 , 9,10. Esta pesquisa conclui que o aumento de glicose no sangue aumenta a atenção e memória, e que esses efeitos são mais fortes quando se trabalha em tarefas cognitivamente exigentes, e nas de idade avançada 1,3. Nas últimas décadas, a investigação sobre o impacto das flutuações de glicose no sangue em psicologia humana também se tornou um tópico de interesse no campo da psicologia social 11-13. Flutuações nos níveis de glicose no sangue têm sido associados a mudanças no humor 14, motivação &lt;sup&gt; 15, auto-controle 11-13, atitudes 16,17, preconceito 18, e à evolução dos no consumidor 19, comendo 20,21, e os domínios financeiros 22 de tomada de decisão. Porque estes resultados transcendem uma grande variedade de variáveis ​​dependentes, a mensagem sobre o papel da glicose no sangue na tomada de decisão é clara: glicose no sangue é a forma essencial da energia para o cérebro, e na abundância de chumbo de glicose no sangue para importantes mudanças na cognição e auto-regulação 11,13,15. Dado o importante papel que a glicose no sangue desempenha em vários resultados psicológicos e comportamentais, o manuscrito atual oferece um método simples para a manipulação de glicose no sangue que os pesquisadores podem usar para testar uma variedade de cognições e comportamentos. Além disso, o manuscrito atual fornece um método para avaliar a acessibilidade cognitiva de palavras ou conceitos-alvo que pode mudar em resposta às flutuações da glicose no sangue usandouma tarefa de decisão lexical. A eficiência dos atuais métodos tornam ideais para pesquisas experimentais, como eles efetivamente manipular glicose no sangue e medir as mudanças cognitivas usando relativamente baratos, produtos comercialmente disponíveis. A acessibilidade do presente manipulação de glicose no sangue faz com que seja mais fácil de executar o grande número de participantes necessários para obter a potência adequada necessária para detectar diferenças estatisticamente significativas no comportamento causados ​​por quaisquer alterações manipulados na glicemia 23. Além disso, a tarefa de decisão lexical é útil na medida em que pode revelar mudanças implícitas na cognição não são facilmente acessados ​​através de medidas mais explícitas, como a auto-relatório examina 24. A tarefa de manipulação de glicose no sangue descrito envolve um período de jejum de 8 horas, seguido pelo consumo dirigido de um calórica ou uma bebida sem calorias 22. Após jejum de oito horas, os participantes são designados aleatoriamente para consumir IET-lhe um açúcar adoçado bebidas ou água. Este método também permite a análise de quaisquer efeitos originais produzidos por adoçantes não calóricos (por exemplo. O aspartame, a sucralose), que sabor doce como adoçantes calóricos tradicionais, mas não contêm os açúcares necessários para elevar os níveis de glicose no sangue 20,22,25 . Se estiver interessado nos efeitos de bebidas não açucaradas calorically, esta tarefa inclui um opcional, 3 rd entre sujeitos condição na qual os participantes consumir uma bebida &quot;zero caloria&quot;. O actual método foi aplicado com sucesso em vários paradigmas psicológicos sociais 12,18,22, e oferece um caminho mais curto para testar os efeitos de alterações na glicemia após a auto-regulação do que as medidas anteriormente utilizados como testes de tolerância à glicose 26. Os autores também fornecem um procedimento para avaliar a acessibilidade cognitiva de palavras-alvo, ou conceitos, usando uma tarefa de decisão lexical 27,28. Neste TAsk, cordas de letras e palavras são apresentadas através do computador em formato de tiro rápido, e geralmente caem em algumas categorias conceituais distintos (na pesquisa descrita, as categorias foram: palavras alto teor de gordura alimentos, palavras de alimentos baixo teor de gordura, não-palavras). Os participantes são convidados a identificar rapidamente (normalmente através de teclas) se uma seqüência de letras (por exemplo, fhens;. De pizza) é uma palavra ou um não-palavra. A velocidade de reação na qual os participantes identificar corretamente carta cordas em cada categoria conceptual mede a relativa facilidade com que o participante é capaz de processar mentalmente palavras nessa categoria conceito. Reações mais rápidas significa que um conceito é mais mentalmente acessíveis e, portanto, &quot;na mente de alguém&quot; 24,29,30. Como consequência, as tarefas de decisão lexical fazer variáveis ​​dependentes úteis, e também pode ser usado como cheques de manipulação eficazes para medir se um conceito recentemente preparado (ver referência 30 para obter mais informações sobre priming) é de fato posteriormente mais Cognitivamente acessível do que seria na ausência de uma plica. Embora a tarefa de decisão lexical é uma tarefa que tem sido utilizado em psicologia social há décadas 29, o manuscrito atual apresenta algumas pequenas variações para o procedimento, um link para um modelo de programa editável 27, e uma lista empiricamente validados de palavras dos autores &#39; trabalho anterior que mede a acessibilidade cognitiva da saúde contra os alimentos não saudáveis ​​20. É a esperança dos autores de que a nova adaptação destes dois procedimentos vai ajudar a levar a inovações nos campos da pesquisa social psicológica, no comportamento alimentar e regulação de energia investigação, e irá facilitar ainda mais a pesquisa comparativa que combina as perspectivas e metodologia de várias disciplinas .

<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.

Os participantes Os métodos acima foram implementados em um estudo executado por Hill e colegas 20 em um tamanho médio, universidade privada do sul dos Estados Unidos. A população de graduação na universidade desde que a amostra de sujeitos e participantes receberam crédito parcial é claro como a compensação para participação no estudo. Usando os métodos exactas descritas no protocolo manuscrito, os aut...

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.

Um método para manipular glicose no sangue e resultando medindo as mudanças na Cognitiva Acessibilidade do Target Estímulos

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

9.0K visualizações.  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.

Vídeo: 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.

Hiperinsulinêmico euglicêmico-Clamp in the Rat Consciente

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

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.

Hiperinsulinêmico euglicêmico-Clamps em Consciente, Ratos Unrestrained

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

Quantitative and Temporal Control of Oxygen Microenvironment at the Single Islet Level

Simultaneous oxygenation and monitoring of glucose stimulus-secretion coupling factors in a single technique is critical for modeling pathophysiological states of islet hypoxia, especially in transplant environments. Standard hypoxic chamber techniques cannot modulate both stimulations at the same time nor provide real-time monitoring of glucose stimulus-secretion coupling factors. To address these difficulties, we applied a multilayered microfluidic technique to integrate both aqueous and gas phase modulations via a diffusion membrane. This creates a stimulation sandwich around the microscaled islets within the transparent polydimethylsiloxane (PDMS) device, enabling monitoring of the aforementioned coupling factors via fluorescence microscopy. Additionally, the gas input is controlled by a pair of microdispensers, providing quantitative, sub-minute modulations of oxygen between 0-21%. This intermittent hypoxia is applied to investigate a new phenomenon of islet preconditioning. Moreover, armed with multimodal microscopy, we were able to look at detailed calcium and KATP channel dynamics during these hypoxic events. We envision microfluidic hypoxia, especially this simultaneous dual phase technique, as a valuable tool in studying islets as well as many ex vivo tissues.

Microfluidic oxygen control confers more than just convenience and speed over hypoxic chambers for biological experiments. Especially when implemented via diffusion through a membrane, microfluidic oxygen can provide simultaneous liquid and gas phase modulations at the microscale-level. This technique enables dynamic multi-parametric experiments critical for studying islet pathophysiology.

Controle quantitativo e temporal de oxigênio Microenvironment no Ilhéu Individual

Simultaneous oxygenation and monitoring of glucose stimulus-secretion coupling factors in a single technique is critical for modeling pathophysiological ...

Bioengenharia

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.

Microinjeção insulina hipocampal e<em&gt; Em vivo</em&gt; Microdiálise Durante um teste de memória espacial

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

Neurociência

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.

Membrana Potencial Dye imagem de Ventromedial Hipotálamo Neurônios de camundongos adultos para estudar 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 ...

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.

Estudando o sinal de insulina hipotalâmico a intolerância de glicose periférica com um sistema de infusão contínua de drogas dentro do cérebro de rato

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.

<meta charset="utf8"></head><body>Aclimatação antes de um teste de tolerância à insulina intraperitoneal para mitigar a hiperglicemia induzida por estresse em camundongos conscientes

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.

Aclimatação antes de um teste de tolerância à insulina intraperitoneal para mitigar a hiperglicemia induzida por estresse em camundongos conscientes

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 ...

Instalação cirúrgica de pinças hiperglicêmicas e hipoglicêmicas em camundongos

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.

Autor em destaque: Investigando a homeostase da glicose no sangue no cérebro murino usando uma técnica de grampo hiperglicêmica e hipoglicêmica econômica

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.

Clamp hiperglicêmico e grampo hipoglicêmico em camundongos conscientes

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

Um método para manipular glicose no sangue e resultando medindo as mudanças na Cognitiva Acessibilidade do Target Estímulos

Resumo

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