SRP409649 PRJNA905102 GSE218709 A subpopulation of lipogenic brown adipocytes drives thermogenic memory [snRNAseq] Sustained responses to transient stimuli are important for animals to survive and reproduce in their environment. However, the mechanisms that underlie altered responses to temporary shifts in abiotic factors, such as temperature, remain poorly understood. Here, we find that transient cold exposure leads to sustained transcriptional and metabolic adaptations in brown adipose tissue, which are critical for an improved thermogenic response to secondary cold encounter. Primary thermogenic challenge triggers the delayed induction of a lipid biosynthesis program even after cessation of the original stimulus, which protects from subsequent exposures. By combining single-nucleus RNA sequencing, spatial transcriptomics, and immunofluorescence imaging, we discover that this lipogenic response is carried out by a distinct subpopulation of brown adipocytes that is localized along the perimeter of classical Ucp1high brown adipocytes. The protective effect of the lipogenic program is associated with the production of acyl carnitines, and supplementation of acyl carnitines recapitulates improved secondary cold responses even in the absence of lipogenesis. Overall, our data highlight the importance of heterogenous brown adipocyte populations for “thermogenic memory” in the setting of repeated cold exposure, which may have implications for therapeutic efforts leveraging short-term thermogenesis to counteract the hypercaloric state of obesity. Overall design: Brown adipose tissue was collected from 5 mice at thermoneutrality and 5 mice that had undergone transient acute cold exposure 4 days previously. Nuclei isolation was then performed as previously described. Nuclei from the 5 mice for each condition were pooled. Sorted nuclei were then immediately encapsulated into droplets, libraries prepared using the 10X Genomics platform and sequencing was performed on NextSeq 550 instrument using 150-cycle kit. BCL files were demultiplexed, aligned to mouse mm10 genome, filtered, and UMI counted using CellRanger (10X Genomics). GSE218709 pubmed 37783943 parent_bioproject PRJNA905100