Objective: The liver releases glucose into the blood using the glucose-6-phosphatase (G6Pase) system, a multiprotein complex located in the endoplasmic reticulum (ER). Here, we show for the first time that the G6Pase system is also expressed in hypothalamic tanycytes, and it is required to regulate energy balance. Methods: Using automatized qRT-PCR and immunohistochemical analyses, we evaluated the expression of the G6Pase system. Fluorescent glucose analogue (2-NBDG) uptake was evaluated by 4D live -cell microscopy. Glucose release was tested using a glucose detection kit and highcontent live -cell analysis instrument, Incucyte s3. In vivo G6pt knockdown in tanycytes was performed by AAV 1 -shG6PT-mCherry intracerebroventricular injection. Body weight gain, adipose tissue weight, food intake, glucose metabolism, c -Fos, and neuropeptide expression were evaluated at 4 weeks post-transduction. Results: Tanycytes sequester glucose -6 -phosphate (G6P) into the ER through the G6Pase system and release glucose in hypoglycaemia via facilitative glucose transporters (GLUTs). Strikingly, in vivo tanycytic G6pt knockdown has a powerful peripheral anabolic effect observed through decreased body weight, white adipose tissue (WAT) tissue mass, and strong downregulation of lipogenesis genes. Selective deletion of G6pt in tanycytes also decreases food intake, c -Fos expression in the arcuate nucleus (ARC), and Npy mRNA expression in fasted mice. Conclusions: The tanycyte-associated G6Pase system is a central mechanism involved in controlling metabolism and energy balance. (c) 2024 The Authors. Published by Elsevier GmbH. This is an open access article under the CC BY -NC license (http://creativecommons.org/licenses/by-nc/4.0/).