Method Article

Infrared Thermography for the Detection of Changes in Brown Adipose Tissue Activity

DOI:

10.3791/64463

September 28th, 2022

In This Article

Summary

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Here, we present a protocol for measuring brown adipose tissue activity after a meal in humans and laboratory animals.

Abstract

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Measuring brown adipose tissue (BAT) activity by positron emission tomography computed tomography (PET-CT) via the accumulation of 18F-fluorodeoxyglucose (FDG) after a meal or in obese or diabetic patients fails as the method of choice. The main reason is that 18F-FDG competes with the postprandial high glucose plasma concentration for the same glucose transporter on the membrane of BAT cells. In addition, BAT uses fatty acids as a source of energy as well, which is not visible with PET-CT and could be changed along with glucose concentration in obese and diabetic patients. Therefore, to estimate the physiological importance of BAT in animals and humans, a new infrared thermography method used in recent publications is applied.

After overnight fasting, BAT activity was measured by infrared thermography before and after a meal in human volunteers and female wild-type mice. The camera software calculates the object's temperature using distance from the object, skin emissivity, reflected room temperature, air temperature, and relative humidity. In mice, the shaved area above the BAT was a region of interest for which average and maximal temperatures were measured. The phase of the estrous cycle in female mice was determined after an experiment by vaginal smears stained with cresyl violet (0.1%) stain solution. In healthy volunteers, two skin areas of the neck were selected: the supraclavicular area (above the collarbone, where BAT cells are present) and the interclavicular area (between the collarbones, where there is no BAT tissue detected). BAT activity is determined by the subtraction of those two values. Also, the average and maximal temperatures of skin areas could be determined in animals and human participants.

Changes in BAT activity after a meal measured by infrared thermography, a non-invasive and more sensitive method, were shown to be sex, age, and phase of the estrous cycle dependent in laboratory animals. As part of diet-induced thermogenesis, BAT activation in humans was also proven to be sex, age, and body mass index dependent. Further determining the pathophysiological changes in BAT activity after a meal will be of great importance for participants with high glucose plasma concentrations (obesity and diabetes mellitus type 2), as well as in different laboratory animals (knock-out mice). This method is also a variable tool for determining possible activating drugs that could rejuvenate BAT activity.

Introduction

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Brown adipose tissue (BAT), in contrast to white adipose tissue (WAT), does not store but rather spends energy. Upon sympathetic stimulation, BAT utilizes fatty acids and glucose and produces heat by the activation of uncoupling protein 1 (UCP1). The function of UCP1 is to use an H+ gradient between two mitochondrial membranes to produce heat instead of ATP. The function of BAT is to increase heat production under cold conditions, which leads to an increase in energy expenditure1. After cold exposure, sensory inputs from the skin inhibit warm-sensitive neurons in the median preoptic (MnPO) nucleus of the hypothalamic preoptic area (P....

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Protocol

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All experimental procedures on laboratory animals were approved by the National Ethical Committee and the Ministry of Agriculture (EP 185/2018). The experiments were conducted in accordance with the Ethical Codex of The Croatian Society for Laboratory Animal Science and ARRIVE guidelines. All procedures performed in studies involving human participants were in accordance with the Declaration of Helsinki and approved by the Ethics Committee of the University of Zagreb, School of Medicine (UP/I-322-01/18-01/56). In this study, we present the results from three female participants (BMI: 29 kg/m2 ± 5 kg/m2). Informed consent was obtained from al....

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Results

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The easiest way to determine BAT activity is to subtract the maximal skin temperature above the BAT before and after a meal in human subjects. A better way to calculate BAT activity is to select two regions of interest: the skin area above the BAT, which is located in the supraclavicular area, and the interclavicular area of the skin where no BAT tissue is found in humans, designated as a reference area (according to PET-CT; Figure 1). BAT activity is then easily determined by the subtractio.......

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Discussion

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Recent studies present growing evidence regarding the physiological regulation and importance of BAT activity in adult humans and animals in the development of obesity and diabetes mellitus. Furthermore, possible BAT activation by exogenous activators is becoming a target for pharmaceutical companies. To be able to estimate the physiological regulation and pathophysiological importance of BAT in very burdensome diseases, as well as discover a potential therapeutic approach, infrared thermography is becoming the method of.......

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Disclosures

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The authors have nothing to disclose.

Acknowledgements

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This study was funded by the Croatian Science Foundation research grant (IP-2018-01- 7416).

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
0.1% cresyl violet acetate Commonly used chemical
Device for measuring air temperature and humidityKesterlKestrel 4200Certificat of conformity
External data storageHard Drive with at least 1 TB
Glass microscopic slidesCommonly used
Small cotton tip swab Urethral swabs
Software for analysisFLIR Systems, Wilsonville, OR, USAFLIR Tools
Software for meassurementsFLIR Systems, Wilsonville, OR, USAResearchIR softwareFLIR ResearchIR Max, version 4.40.12.38 (64-bit)
Thermac CameraFLIR Systems, Wilsonville, OR, USAFLIR T-1020

References

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  1. van Marken Lichtenbelt, W. D., et al. Cold-activated brown adipose tissue in healthy men. New England Journal of Medicine. 360 (15), 1500-1508 (2009).
  2. Morrison, S. F., Nakamura, K. Central neural pathways for thermoregulation. Frontiers in Bioscience. 16....

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Tags

Infrared ThermographyBrown Adipose TissueBAT ActivityNoninvasive Temperature MeasurementSupraclavicular AreaPostprandial BAT ActivationEstrous Cycle AnalysisSkin TemperatureObesity Diabetes ResearchThermal Imaging Protocol

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