library_construction_protocol: Mice were anesthetized with isoflurane and then rapidly sacrificed by decapitation. Brains were extracted and sliced at 225-μm thickness using a vibratome. Slices were taken in an ice-cold high-sucrose slicing solution consisting of the following components (in mM): 87 NaCl, 75 sucrose, 25 glucose, 25 NaHCO3, 1.25 NaH2PO4, 2.5 KCl, 7.5 MgCl2, 0.5 CaCl2 and 5 ascorbic acid saturated with 95% O2/5% CO2. Slices were then enzyme-treated for ~15 min at 34°C with protease XXIII (2.5 mg/mL; Sigma) in a high-sucrose dissociation solution containing the following (in mM): 185 sucrose, 10 glucose, 30 Na2SO4, 2 K2SO4, 10 HEPES buffer, 0.5 CaCl2, 6 MgCl2, 5 ascorbic acid (pH 7.4) and 319 mOsm. Slices were washed three times with cold dissociation solution then transferred to a trypsin inhibitor/bovine serum albumin (TI/BSA) solution (10 mg/mL; Sigma) in cold dissociation solution. 7-8 slices containing the MPOA and 9-10 slices containing the ARC were obtained per group. MPOA and ARC were microdissected using ultra fine forceps and prepared for sequencing. Microdissected tissue was kept in cold TI/BSA dissociation solution until trituration. Immediately before dissociation, tissue was incubated for ~10 min at 37°C, then triturated with a series of small bore fire-polished Pasteur pipettes. Cell-suspension was kept on ice until single-cell capture. Cell viability was measured for each sample preparation using a Countess II automated cell counter (ThermoFisher) and was between the range of 67.5% to 82.3% before processing with the 10x Genomics platform61. 12,000 cells were loaded for capture onto an individual lane of a Chromium Controller (10x Genomics) for each sample. Single-cell capture, barcoding, and library preparation were performed using the 10x Chromium platform according to the manufacturer's protocol (#CG00052) using version 3 (V3) chemistry. The single cell suspension was obtained. The cells were counted and examined for viability with trypan blue staining using a TC-20 cell counter (Bio-Rad). About 12500 cells at average of 5×105 cells/ml live cell concentration with about 70% viability were loaded into the Single Cell Chip G to generate single cell emulsion in Chromium Controller with Chromium Single Cell 3' Library & Gel Bead Kit v3.1 (10x Genomics, Cat. 1000268). Reverse transcription of mRNA and cDNA amplification were carried out following the manufacture's instruction (10x Genomics, Cat. 1000268). The amplified cDNA was further fragmented to construct NGS libraries. The libraries were then sequenced on NovaSeq 6000 sequencer (Illumina) by the NIEHS Epigenomics and DNA Sequencing Core Laboratory with the parameters recommended in the manufacture's instruction. Total 40,000 reads/cell were achieved for each sample.