The snRNAseq Analysis of JunB deficient brown adipose tissue and control samples

The coexistence of brown adipocytes with low and high thermogenic activity is a fundamental feature of brown adipose tissue (BAT) heterogeneity and plasticity. However, the mechanisms that govern thermogenic adipocyte heterogeneity and its significance in obesity and metabolic disease remain poorly understood. Here we show that in male mice, a population of JunB-enriched (JunB+) adipocytes within the BAT exhibits lower thermogenic capacity compared to high-thermogenic adipocytes. To determine if JunB plays a role in the control of brown adipocyte heterogeneity in thermogenesis, we performed single-nucleus RNA sequencing (snRNA-seq) of BAT from JunB FKO and control mice (3-month-old male mice (5/group)) housed at room temperature. Our study pointing to a potent inhibition by JunB in the conversion from low- to high-thermogenic adipocyte. The nuclei was isolated from newly collected brown fat tissue of 10-week-old JunB deficient and control mice (n=5/group). The same number of nuclei from each individual animal were combined within a group and used for snRNAseq.棕色脂肪细胞同时具备低产热活性与高产热活性两种表型，是棕色脂肪组织（brown adipose tissue, BAT）异质性与可塑性的核心特征之一。然而，调控产热脂肪细胞异质性的分子机制，及其在肥胖与代谢疾病中的生物学意义，目前仍未被充分阐明。本研究以雄性小鼠为研究对象，发现棕色脂肪组织中一类富集JunB（JunB+）的脂肪细胞群体，其产热能力低于高产热脂肪细胞。为明确JunB是否参与调控棕色脂肪细胞的产热异质性，我们对饲养于室温环境下的JunB FKO与对照雄性小鼠（3月龄，每组5只）的棕色脂肪组织开展了单细胞核RNA测序（single-nucleus RNA sequencing, snRNA-seq）。此外，我们从10周龄JunB缺陷型与对照雄性小鼠的新鲜棕色脂肪组织中分离细胞核，将每只个体动物的等量细胞核混合分组，用于后续snRNA-seq实验。本研究结果提示，JunB可有效抑制低产热脂肪细胞向高产热脂肪细胞的转化。