Dissection, histological processing, and gene expression analysis of murine supraclavicular brown adipose tissue

Authors: WATERSTRAAT, MARK - Rice University; WANG, ZIYI - Children'S Nutrition Research Center (CNRC); KOGISO, MARI - Children'S Nutrition Research Center (CNRC); CABALLERO-JUAREZ, ROMMEL - Rice University; CHEN, MIAO-HSUEH - Children'S Nutrition Research Center (CNRC)

Submitted to: The Journal of Visualized Experiments (JoVE)
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/10/2024
Publication Date: 3/29/2024
Citation: Waterstraat, M.G., Wang, Z., Kogiso, M., Caballero-Juarez, R., Chen, M. 2024. Dissection, histological processing, and gene expression analysis of murine supraclavicular brown adipose tissue. The Journal of Visualized Experiments (JoVE). 205. Article e66475. https://doi.org/10.3791/66475.
DOI: https://doi.org/10.3791/66475

Interpretive Summary: Brown fat is a special type of fat that helps control how our bodies use energy. This type of fat was recently found in the necks of healthy adult humans and mice. However, studying this newly identified brown fat can be difficult because of its small size and location. In this paper, we report new methods to study this neck brown fat more effectively. These methods could assist researchers in understanding the functions of neck brown fat and its role in managing our body’s metabolism.

Technical Abstract: Brown adipose tissue (BAT)-mediated thermogenesis plays an important role in the regulation of metabolism, and its morphology and function can be greatly impacted by environmental stimuli in mice and humans. Currently, murine interscapular BAT (iBAT), which is located between two scapulae in the upper dorsal flank of mice, is the main BAT depot used by research laboratories to study BAT function. Recently, a few previously unknown BAT depots were identified in mice, including one analogous to human supraclavicular brown adipose tissue. Unlike iBAT, murine supraclavicular brown adipose tissue (scBAT) is situated in the intermediate layer of the neck and thus cannot be accessed as readily. To facilitate the study of newly identified mouse scBAT, presented herein is a protocol detailing the steps to dissect intact scBAT from postnatal and adult mice. Due to scBAT's small size relative to other adipose depots, procedures have been modified and optimized specifically for processing scBAT. Among these modifications is the use of a dissecting microscope during tissue collection to increase the precision and homogenization of frozen scBAT samples to raise the efficiency of subsequent qPCR analysis. With these optimizations, the identification of, morphological appearance of, and molecular characterization of the scBAT can be determined in mice.