Our objective was to investigate the effect of propionate on the hypothalamic regulation of food intake in the context of the hepatic oxidation theory in ruminants. Nine Rambouillet lambs (27.93 ± 4.6 kg) were randomly assigned to one of 3 experimental diets (n = 3/diet): T1, a control diet containing 93% alfalfa hay and 7% molasses, without supplementation of Calcium-Propionate (CaPr); T2, T1 + 30 g/kg DM of CaPr; and T3, T1 + 35 g/kg DM of CaPr. Diets were offered ad libitum as a total mixed ration. The lambs were housed in individual cages equipped with feed and water bowls. Lambs were first adapted to their diets (without CaPr) for 10 d, and the experimental period lasted 42 d (lambs will receive treatments during this period). On d 42, lambs were euthanized, the hypothalamus collected, and RNA was extracted and sequenced on the Illumina HiSeq 4000 system, generating an average of 50 million reads/sample, of which 75% where uniquely mapped, with 56% of gene-assigned read, with a total of 15608 uniquely identified genes. A linear model with CaPr as fixed and animal as random effect was fitted. Differentially expressed genes (DEG) were declared at fold change ≥ |2| and P-value ≤0.05. The Dynamic Impact Approach was used for pathway analyses using the KEGG database. Two contrast yielded the highest number of DEG: T2+T3vsT1 (overall effect of CaPr infusion vs control), and T3vsT1 (3.5% CaPr vs control), with 1200 and 919 DEG, respectively. Independently of the contrast, pathway analysis revealed a broad downregulation of all pathways in animals that received CaPr. The upregulated pathways in CaPr lambs compared with control (T3vsT1) were galactose, glyoxylate and dicarboxylate, β-alanine, and porphyrin metabolism, terpenoid backbone biosynthesis, peroxisome, ferroptosis, and one carbon pool by folate. In addition, ether lipids and sphingolipid metabolism, mucin type O-glycan and heparan sulfate glycosaminoglycan biosynthesis were upregulated in both contrasts. The transcriptome analysis highlighted an extended downregulation of all hypothalamic activities in ewes that received CaPr infusions, independently of the dose. Further work is needed to assess the hypothalamic mechanisms involved in feed intake regulation.