Abstract
The Brown Adipose Tissue (BAT) is composed by mitochondrial rich, multilocular adipocytes, in strict topographical and functional relation with vasculature and noradrenergic nerves. Brown adipocytes are able to dissipate energy to produce heat, in a process known as non-shivering thermogenesis. Due to its contribution to energy expenditure, BAT is intensely studied for its potential to counteract metabolic diseases such as obesity, type 2 diabetes, dyslipidemia and cardiovascular diseases. BAT displays specific morphological characteristics that allow to assess its functional state. In this chapter we describe methodologies to properly dissect BAT depots, evaluate their gross anatomy, and assess its activation by light microscopy using peroxidase immunostaining and by laser scanning confocal microscopy using immunofluorescence. We also describe methodologies to study BAT ultrastructure by transmission and scanning electron microscopy, to visualize peroxidase immunostaining reactions at an ultrastructural level and to perform immunofluorescence reactions on paraffin-embedded samples, more often available in the clinical setting (due to the possibility to store them long-term) as opposed to fresh samples. The described techniques can be employed to study BAT morphology and activation in response to various stimuli (e.g., cold exposure; specific dietary composition) and in different pathological conditions (e.g., obesity; type 2 diabetes).
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Acknowledgments
This work was supported by grants from the Italian Ministry of University (PRIN 2017, # 2017L8Z2EM). We thank Professor Shingo Kajimura from Harvard University for providing Ucp1Cre/+; Rosa26mT/mG reporter mice. We thank Professors Harold Sacks from the VA of the Los Angeles Healthcare System, Professor Orian Shiriai from University of California Los Angeles, and Doctor Anton Petcherski and Doctor Rebeca Acın-Perez from David Geffen School of Medicine, University of California for providing human adipose tissue samples of a patient suffering from pheochromocytoma. We thank Professor Antonio Osculati from the University of Pavia for providing human BAT samples.
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Colleluori, G., Perugini, J., Di Vincenzo, A., Senzacqua, M., Giordano, A., Cinti, S. (2022). Brown Fat Anatomy in Humans and Rodents. In: Guertin, D.A., Wolfrum, C. (eds) Brown Adipose Tissue. Methods in Molecular Biology, vol 2448. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2087-8_2
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DOI: https://doi.org/10.1007/978-1-0716-2087-8_2
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