The vestibular system controls balance, posture, blood pressure, and gaze. However, the roles of the vestibular system in energy and glucose metabolism remain unknown. We herein examined the roles of the vestibular system in obesity and impaired glucose metabolism using mice with vestibular lesions (VL) fed a high-sucrose/high-fat diet (HSHFD). VL was induced by surgery or arsenic. VL significantly suppressed body fat enhanced by HSHFD in mice. Glucose intolerance was improved by VL in mice fed HSHFD. VL blunted the levels of adipogenic factors and pro-inflammatory adipokines elevated by HSHFD in the epididymal white adipose tissue of mice. A β-blocker antagonized body fat and glucose intolerance enhanced by HSHFD in mice. The results of an RNA sequencing analysis showed that HSHFD induced alterations in genes, such as insulin-like growth factor-2 and glial fibrillary acidic protein, in the vestibular nuclei of mice through the vestibular system. In conclusion, we herein demonstrated that the dysregulation of the vestibular system influences an obese state and impaired glucose metabolism induced by HSHFD in mice. The vestibular system may contribute to the regulation of set points under excess energy conditions. OPEN ACCESS Citation: Kawao N, Takafuji Y, Ishida M, Okumoto K, Morita H, Muratani M, et al. (2020) Roles of the vestibular system in obesity and impaired glucose metabolism in high-fat diet-fed mice. PLoS ONE 15 (2): e0228685. https://doi.org/10.sympathetic nervous system in rodents [4,5]. Luxa et al. reported that labyrinthectomy increased myofiber remodeling in the soleus muscle of mice [6]. We recently revealed that gravity changes affect muscle and bone through the vestibular system in mice [7,8]. Collectively, these findings suggest that the vestibular system regulates the musculoskeletal system partly through the sympathetic nervous system. Dysfunctions in the vestibular system clinically cause dizziness, vertigo, and unsteadiness [9]. Long-term space flight impairs the vestibular system in astronauts who experience orthostatic intolerance and unsteadiness [3]. However, the roles of the vestibular system in metabolic homeostasis have not yet been elucidated in detail.Excess energy is stored in white adipose tissue (WAT) as lipids and causes obesity, a risk factor for diabetes [10]. Adipocytes differentiate from mesenchymal stem cells, and the activation of adipogenic differentiation is followed by enhanced levels of peroxisome proliferatoractivated receptor γ (PPARγ), aP2, long chain acyl-CoA synthetase (ACSL) 1, and lipoprotein lipase (LPL). In obesity, WAT releases pro-inflammatory adipokines, such as tumor necrosis factor (TNF)-α, plasminogen activator inhibitor (PAI)-1, and monocyte chemoattractant protein (MCP)-1, which impair glucose metabolism due to the induction of low-grade systemic inflammation [10]. Adiponectin and leptin released from adipose tissue exert pleiotropic effects in glucose metabolism [10]. Circulating adiponectin and leptin produced from WAT are negatively and positively r...