Defective nutrient storage and adipocyte enlargement (hypertrophy) are emerging features of metabolic syndrome and type 2 diabetes. How the cytoskeletal network contributes to nutrient uptake, fat storage, and adipocyte size remains poorly understood. Utilizing the Drosophila larval fat body (FB) as a model adipose tissue, we show that a specific actin isoform—Act5C—forms the cortical actin network necessary for inter-organ lipid trafficking. Act5C also promotes FB tissue expansion during larval development so larvae can store sufficient biomass for metamorphosis. We find FB-specific loss of Act5C, but not other Drosophila actins, perturbs FB triglyceride (TG) storage in lipid droplets (LDs), resulting in developmentally delayed larvae that fail to develop into flies. Act5C localizes to the FB cell surface where it intimately contacts peripheral LDs (pLDs), forming a cortical actin network together with spectrins for cell architectural support. While both the cortical actin and spectrin cytoskeletons maintain FB cell surface architecture, we find that only the actin network is required for fat storage. Mechanistically, we show that FBs lacking the Act5C cortical cytoskeleton exhibit a block in lipoprotein (Lpp) secretion from FB cells, and a subsequent disruption of gut:FB inter-organ lipid transport, resulting in mid-gut fat accumulation. Utilizing temporal RNAi-depletion approaches, we also reveal that Act5C is indispensable post-embryogenesis during larval feeding to promote FB cell expansion. Act5C-deficient FBs fail to expand cell sizes, leading to lipodystrophic larvae unable to accrue sufficient biomass for metamorphosis. Collectively, we propose that the Act5C-mediated cortical actin network of Drosophila adipose tissue plays an essential role in post-embryonic inter-organ nutrient transport and FB cell size determination for organismal energy homeostasis and development.

These are datasets associated with cell size quantification.