Chain length defines the spatial segregation and metabolic fate of fatty acids in adipocytes

Adipocytes primarily store fatty acids (FAs) as triacylglycerols (TGs) within lipid droplets, releasing them through lipolysis to meet systemic energy demands. While the metabolism of long-chain fatty acids (LCFAs) in adipocytes has been extensively characterized, it remains ill-defined how adipocytes utilize fatty acids depending on chain length and structure. Our work demonstrates that short- and medium-chain fatty acids (SMCFAs) are esterified into TGs within lipid droplets, rather than being incorporated into other FA-containing lipid species. During lipolytic activation, TGs enriched in SMCFAs are hydrolysed more rapidly than those containing LCFAs. This accelerated mobilization is facilitated by the preferential localization of SMCFA-containing TGs at the lipid droplet surface, which enhances accessibility to adipose triglyceride lipase. Unlike LCFAs, which are efficiently released for utilization by peripheral tissues, SMCFAs are predominantly oxidized within adipocytes. These findings reveal a unique metabolic routing of SMCFAs, indicating their preferential intracellular oxidation to support adipocyte energy requirements.