To isolate RNA, each hypothalamus tissue sample was placed in a 1.5 mL centrifuge tube containing a 3 mm steel bead (Fisher, 50-206-7603). Subsequently, 350 µL of RLT Plus buffer (Qiagen) supplemented with β-mercaptoethanol was added to the tube. Homogenization was performed using a Tissuelyser II (Qiagen) for two cycles at 25 Hz for 3 minutes each. Following homogenization, samples were centrifuged at 16,000 × g for 3 minutes, and the clarified lysates were collected. RNA isolation was then carried out according to the manufacturer's protocol using the RNeasy Plus Micro Kit (Qiagen, Cat. No. 74034). Purified RNA was transferred to 1.5 mL DNA LoBind tubes (Eppendorf), snap-frozen on dry ice, and stored at −80 °C until further processing for RNA sequencing. For each of the 18 independent biological samples, a sequencing library was prepared with the QuantSeq 3' mRNA-seq Library Prep Kit FWD for Illumina (Lexogen A01173). The UMI second strand synthesis module was used to introduce unique molecular identifiers (UMIs), following the protocol defined by the manufacturer.  Briefly, oligo dT primers were hybridized to total RNA for reverse transcription, followed by RNA removal after first strand synthesis. UMIs were introduced during second strand synthesis and cDNA was purified using magnetic beads, followed by 18 cycles of PCR with dual indexes and PCR purification. Library quantity and quality were assessed with Qubit™ 1X dsDNA HS Assay Kits (Invitrogen Q33231) and Bioanalyzer High Sense DNA chip (Agilent 5067-4626). Libraries were pooled in equimolar ratios, and the sequencing pool was purified with magnetic beads to remove residual adaptor dimers.