The peroneocuboid joint, between the peroneus longus tendon and the cuboid bone, has not been anatomically well-defined and no embryological study has been published. Furthermore, the ossification of the os peroneum (a sesamoid inside the peroneus longus tendon) and its associated pathology has been considered to be generated by orthostatic and/or mechanical loads. A light microscopy analysis of serially sectioned human embryonic and fetal feet, the analysis of human adult feet by means of standard macroscopic dissection, X-ray and histological techniques have been carried out. The peroneus longus tendon was fully visible until its insertion in the 1st metatarsal bone already at embryonic stage 23 (56–57 days). The peroneocuboid joint cavity appeared at the transition of the embryonic to the fetal period (8–9th week of gestation) and was independent of the proximal synovial sheath. The joint cavity extended from the level of the calcaneocuboid joint all the way to the insertion of the peroneus longus tendon in the 1st metatarsal bone. The frenular ligaments, fixing the peroneus longus tendon to the 5th metatarsal bone or the long calcaneocuboid ligament, developed in the embryonic period. The peroneus longus tendon presented a thickening in the area surrounding the cuboid bone as early as the fetal period. This thickening may be considered the precursor of the os peroneum and was similar in shape and in size relation to the tendon, to the os peroneum observed in adults. To the best of our knowledge, this is the first study to show that the os peroneum, articular facets of the peroneus longus tendon and cuboid bone, the peroneocuboid joint and the frenular ligaments appear during the embryonic/fetal development period and therefore they can not be generated exclusively by orthostatic and mechanical forces or pathological processes.