Name: a method for manipulating blood glucose and measuring resulting changes in cognitive accessibility of target stimuli
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Esta investigación se realizó con fondos de subsidios de la Fundación Anthony M. Marchionne (70256 hasta 23284) y TCU. Los autores agradecen a Danielle DelPriore, Amanda Morin, y Christopher Rodeheffer por sus útiles contribuciones a la formación de estos métodos. Los autores también desean agradecer a Hannah Bradshaw y Randi Proffitt Leyva por su ayuda con la filmación de este protocolo.

<ol>
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Los autores no tienen nada que revelar.

Este manuscrito describe un procedimiento simple, barato para la manipulación de la glucosa en sangre, así como un procedimiento para la medición de los cambios resultantes en la accesibilidad cognitiva de palabras y conceptos de los objetivos. Los métodos descritos anteriormente pueden aplicarse a una amplia gama de áreas de investigación incluyendo la psicología social, la psicología cognitiva, y ciencias de la nutrición. Teniendo en cuenta que este método puede ser utilizado por personas en múltiples área...

Los investigadores de la psicología cognitiva y la neurociencia han estudiado durante mucho tiempo el impacto de las fluctuaciones de glucosa en sangre en el cerebro, la cognición y el comportamiento 1. Por ejemplo, los investigadores han encontrado que las fluctuaciones de glucosa en la sangre predicen diferencias en la memoria (verbales, retención de dígitos, de trabajo, y episódica) 1-5, la atención 6,7, la solución de rompecabezas 8, y el rendimiento en las tareas de diferentes niveles de demanda cognitiva 2 , 9,10. Esta investigación concluye que el aumento de glucosa en la sangre aumenta la atención y la memoria, y que estos efectos son más fuertes cuando se trabaja en tareas cognitivamente exigentes, y en los de más edad de 1,3 años. En las últimas décadas, la investigación sobre el impacto de las fluctuaciones de glucosa en sangre en la psicología humana también se ha convertido en un tema de interés en el campo de la psicología social 11-13. Las fluctuaciones en la glucosa en sangre se han relacionado con cambios en el estado de ánimo 14, motivación &lt;sup&gt; 15, el autocontrol 11-13, actitudes 16,17, 18 perjuicio, ya los cambios en la toma de decisiones en los consumidores 19, 20,21 de comer, y los dominios financieros 22. Debido a que estos resultados trascienden una amplia variedad de variables dependientes, el mensaje sobre el papel de la glucosa en sangre en la toma de decisiones es clara: la glucosa en sangre es la forma fundamental de energía para el cerebro, y los agotamientos de plomo de glucosa en sangre a importantes cambios en la cognición y autorregulación 11,13,15. Teniendo en cuenta el importante papel que desempeña la glucosa en sangre en varios resultados psicológicos y de comportamiento, el manuscrito actual ofrece un método sencillo para la manipulación de glucosa en la sangre que los investigadores pueden utilizar para probar una gama de conocimientos y comportamientos. Además, el manuscrito actual proporciona un método para evaluar la accesibilidad cognitiva de palabras o conceptos de los objetivos que pueden cambiar en respuesta a las fluctuaciones de la glucosa en sangre usandouna tarea de decisión léxica. La eficacia de los presentes métodos los hacen ideales para la investigación experimental, ya que manipular con eficacia la glucosa en sangre y medir los cambios cognitivos utilizando productos relativamente baratos, disponibles comercialmente. La asequibilidad de la manipulación de glucosa presente en la sangre hace que sea más fácil para ejecutar el gran número de participantes necesarios para obtener la potencia adecuada necesaria para detectar diferencias estadísticamente significativas en el comportamiento causado por los cambios en la glucosa sanguínea manipulados 23. Además, la tarea de decisión léxica es útil porque puede revelar cambios implícitos en la cognición no se accede fácilmente a través de medidas más explícitas como la auto-informe de 24 encuestas. La tarea de manipulación de glucosa en sangre descrito implica un período de ayuno de 8 horas, seguido por el consumo de calorías dirigida de una o una bebida sin calorías 22. Después de un ayuno de ocho horas, los participantes son asignados al azar a consumir EITella un azúcar endulzado bebidas o agua. Este método también permite el ensayo de cualquier efecto únicos producidos por edulcorantes no calóricos (por ejemplo. Aspartamo, sucralosa), que tienen un sabor dulce como edulcorantes calóricos tradicionales, pero no contienen los azúcares necesarios para elevar los niveles de glucosa en la sangre 20,22,25 . Si está interesado en los efectos de bebidas azucaradas no calórico, esta tarea incluye un opcional, 3ª entre sujetos condición en la que los participantes consumen una bebida &quot;cero calorías&quot;. El método actual se ha aplicado con éxito en varios paradigmas psicológicos sociales 12,18,22, y ofrece un camino más corto para probar los efectos de los cambios en la glucosa sanguínea en la autorregulación que las medidas utilizadas anteriormente como prueba de tolerancia de glucosa 26. Los autores también proporcionan un procedimiento para evaluar la accesibilidad cognitiva de las palabras objetivo, o conceptos, utilizando una tarea de decisión léxica 27,28. En este task, cadenas de letras y palabras se presentan a través del ordenador en formato de tiro rápido, y normalmente se dividen en varias categorías conceptuales distintas (en la investigación descrita, las categorías fueron: palabras de alto contenido graso de los alimentos, palabras de alimentos bajos en grasa, no palabras). Se les pide a los participantes que identifiquen rápidamente (normalmente a través de pulsación de tecla) si una cadena de letras (por ejemplo; fhens. Pizza) es una palabra o una no-palabra. La velocidad de reacción en la que los participantes identifiquen correctamente cadenas de letras en cada categoría conceptual mide la facilidad relativa con la que el participante es capaz de procesar mentalmente las palabras de esa categoría concepto. Reacciones más rápidas significan que un concepto es mentalmente más accesible, y por lo tanto, &quot;en la mente de alguien&quot; 24,29,30. Como consecuencia, las tareas de decisión léxica hacen variables dependientes útiles, y también pueden ser utilizados como controles eficaces de manipulación para medir si un concepto recientemente preparado (véase la referencia 30 para más información sobre el cebado) es, en efecto, posteriormente, más Cognitivamente accesible de lo que sería en ausencia de un número primo. Aunque la tarea de decisión léxica es una tarea que se ha utilizado en la psicología social durante décadas 29, el manuscrito actual presenta algunas variaciones menores en el procedimiento, un enlace a una plantilla de programa editable 27, y una lista validada empíricamente de las palabras de los autores el trabajo previo que mide la accesibilidad cognitiva de la salud frente a los alimentos no saludables 20. Es la esperanza de los autores de que la adaptación adicional de estos dos procedimientos ayudará a conducir a las innovaciones en el campo de la investigación psicológica social, de la conducta alimentaria y la regulación de la energía de investigación y facilitará aún más la investigación comparativa que combina las perspectivas y metodología de múltiples 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.

Participantes Los métodos anteriores se llevaron a cabo en un estudio dirigido por Hill y colegas 20 a una de tamaño mediano, universidad privada en el sur de Estados Unidos. La población universitaria de la Universidad proporcionó la muestra de sujetos y los participantes recibieron el crédito del curso parcial como compensación por la participación en el estudio. El uso de los métodos exactos descritos en ...

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 método para la manipulación de glucosa en la sangre y la medición de los cambios resultantes en Cognitiva Accesibilidad del destino 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

8.9K vistas.  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.

Hiperinsulinémico euglucémico pinza en la rata consciente

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

Investigación

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.

Hiperinsulinémico euglucémico abrazaderas en ratones conscientes, sin freno

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.

Microinyección insulina hipocampo y<em&gt; En vivo</em&gt; Microdiálisis durante la prueba de memoria espacial

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

Neurociencias

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.

Potencial de Membrana Tinte Imaging de ventromedial del hipotálamo neuronas de ratones adultos para el Estudio de la glucosa 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.

Estudio de la señal de insulina hipotalámica a la intolerancia de la glucosa periférica con un sistema de infusión continua de medicamentos en el cerebro de ratón

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.

Uso de una combinación de calorimetría indirecta, termografía infrarroja y niveles de glucosa en sangre para medir la termogénesis del tejido adiposo marrón en humanos

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

Instalación quirúrgica de pinzas hiperglucémicas e hipoglucémicas en ratones

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 destacado: Investigación de la homeostasis de la glucosa en sangre en el cerebro murino utilizando una técnica de pinza hiperglucémica e hipoglucémica rentable

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 hiperglucémica y pinza hipoglucémica en ratones conscientes

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

Un método para la manipulación de glucosa en la sangre y la medición de los cambios resultantes en Cognitiva Accesibilidad del destino Estímulos

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Capítulos en este video

Hiperinsulinémico euglucémico pinza en la rata consciente

Hiperinsulinémico euglucémico abrazaderas en ratones conscientes, sin freno

Microinyección insulina hipocampo y

Potencial de Membrana Tinte Imaging de ventromedial del hipotálamo neuronas de ratones adultos para el Estudio de la glucosa Sensing

Estudio de la señal de insulina hipotalámica a la intolerancia de la glucosa periférica con un sistema de infusión continua de medicamentos en el cerebro de ratón

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

Uso de una combinación de calorimetría indirecta, termografía infrarroja y niveles de glucosa en sangre para medir la termogénesis del tejido adiposo marrón en humanos

Pinza hiperglucémica y pinza hipoglucémica en ratones conscientes

Pinza hiperglucémica y pinza hipoglucémica en ratones conscientes

Pinza hiperglucémica y pinza hipoglucémica en ratones conscientes