Ependymoma is a rare tumour of the central nervous system that can vary in anatomical location and produce a broad range of clinical signs. Sporadically described in dogs, cats, cattle and horses, this disease has never previously been reported in a mule. This report describes a case of a 12-year-old mule showing neurological signs, including blindness, dysphagia, incoordination, sternal recumbency, depression, apathy and emaciation. Because of the worsening health conditions, the animal was subjected to euthanasia for humane reasons. At necropsy, an area of malacia in the right subcortical region of the frontoparietal area of the cerebrum and an ovoid, firm, grey, 2.4 × 1.0 × 0.7 cm nodule in the right lateral ventricle was observed. Histopathology revealed that the nodule was a densely cellular mass with cells arranged in nests and cords, formations of pseudorosettes with perivascular deposition of fibrillar eosinophilic material and the presence of a few lymphocytes. The cells had rounded and moderately hyperchromatic nuclei, punctate chromatin, distinct nucleoli, and indistinct cytoplasmic margins. Based on the histopathological features, the nodule was diagnosed as an ependymoma, a tumour considered rare, but that may be included in the differential diagnosis for diseases with neurological signs.