biostudies-arrayexpressUnknownTranscriptomicsGenomicsProteomicsTongtong WangIllumina NovaSeq 6000single nucleus RNA sequencingMus musculusMus musculushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-17074Brown adipose tissue (BAT) regulates whole-body energy balance through UCP1-dependent thermogenesis and secretion of metabolic factors. Our single-nucleus RNA sequencing of BAT from cold-exposed Ucp1 knockout mice reveals a distinct brown adipocyte subpopulation (U2). U2 adipocytes exhibit a secretory profile enriched in batokines like growth differentiation factor 15 (GDF15), suggesting a shift towards an endocrine role. Functional analyses reveal that GDF15-GFRAL signaling is required to sustain energy expenditure in adipose tissue (AT). Additionally, a conserved UCP1-GDF15 regulatory axis in human AT is observed.biostudies-arrayexpressSequencing - All sequencing libraries, including mouse single-nucleus libraries and human deep neck tissue bulk RNA-seq libraries, were sequenced on an Illumina NovaSeq 6000 instrument.Library Construction - 10X Genomics libraries for single-nucleus RNA-seq were prepared with the Chromium Single Cell V3.1 reagent kit following the manufacturer’s protocol. Nuclei suspensions (~1200 nuclei per μl) were loaded into Chip G followed by reverse transcription to obtain cDNA, which was subsequently amplified and used for library construction. Human tissue bulk RNA-seq libraries were prepared using standard institutional protocols at FGCZ.Nucleic Acid Extraction - For snRNA-seq, mature adipocyte nuclei were isolated by homogenizing minced tissue in 0.1% CHAPS in CST buffer on ice, lysed for 5 minutes, filtered through a 40 μm cell strainer, and purified via FACS sorting based on RFP fluorescence. For RT-qPCR and bulk RNA-seq, total RNA was extracted using Trizol reagent (Invitrogen) followed by DNase (NEB BioLabs) digestion to remove genomic DNA contamination.Sample Collection - Wild-type (WT) and Ucp1-knockout (Ucp1 KO) mice were euthanized at the study endpoint, and metabolic tissues including interscapular brown adipose tissue (iBAT) and inguinal white adipose tissue (iWAT) were excised and collected. For human samples, deep neck adipose tissue biopsies were acquired from 45 individuals. For single-nucleus RNA sequencing, 300-400 mg of fresh or frozen mouse adipose tissue was immediately minced into 1-3 mm pieces on ice for nuclear isolation.OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesTongtong WangfalseA cold-induced GDF15-secreting adipocyte subpopulation regulates energy homeostasis through endocrine signalingBrown adipose tissue (BAT) regulates whole-body energy balance through UCP1-dependent thermogenesis and secretion of metabolic factors. Our single-nucleus RNA sequencing of BAT from cold-exposed Ucp1 knockout mice reveals a distinct brown adipocyte subpopulation (U2). U2 adipocytes exhibit a secretory profile enriched in batokines like growth differentiation factor 15 (GDF15), suggesting a shift towards an endocrine role. Functional analyses reveal that GDF15-GFRAL signaling is required to sustain energy expenditure in adipose tissue (AT). Additionally, a conserved UCP1-GDF15 regulatory axis in human AT is observed.2026-06-11T00:00:00Z2026-06-11T01:00:44.629Z2026-05-26T10:39:46.654ZE-MTAB-17074ERP193815EFO_0002944EFO_0004170EFO_0009809EFO_0005518EFO_0004184