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Equine Gastric Ulcer Syndrome is a highly prevalent condition, yet its clinical diagnosis remains challenging due to the nonspecific nature of many associated signs. The relationship between the anatomical location of gastric ulcers and clinical presentation is still poorly defined. This retrospective study aimed to evaluate the association between gastric ulcer location and the clinical signs observed in horses. Medical records of 52 horses diagnosed with gastric ulcers by gastroscopy were retrospectively reviewed. Clinical signs were classified as physical, behavioral, or mixed, and ulcer localization was categorized as Equine Squamous Gastric Disease (ESGD), Equine Glandular Gastric Disease (EGGD), or mixed. Associations between variables were analyzed using chi-square or Fisher’s exact tests, with p < 0.05 considered significant. A significant association was identified between the type of clinical signs and ulcer location (p < 0.001). Physical clinical signs were predominantly associated with ESGD, whereas behavioral and mixed signs were more frequently observed in horses with EGGD or mixed ulceration. Furthermore, both the type of clinical signs and ulcer location were associated with the horse’s sport activity. No association was found between ulcer severity and clinical signs nor between ulcer location and sex. These findings suggest that gastric ulcer location influences clinical presentation, indicating that ESGD and EGGD may be associated with different clinical sign patterns.