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Appendicular skeletons of isotemnid notoungulates are described from Cañadón Vaca (Vacan “subage”, Casamayoran South American Land Mammal “Age”, ?middle to late Eocene). Simpson documented three of these, Thomashuxleya externa, Anisotemnus distentus, and Pleurostylodon similis, some 70 years ago, in fashioning a composite isotemnid skeleton, but he did not emphasize their differences from one another. We note variation, especially in the forelimb, that appears to be functionally significant as well as phylogenetically informative. For example, the downwardly curved olecranon, ventrally concave bowing of the ulnar shaft, and orthogonally directed articulation of the elbow joint suggest an erect forelimb stance in Thomashuxleya externa, whereas the forelimbs of Anisotemnus distentus and Pleurostylodon similis show indications of a crouching posture, including ventrally convex bowing of the ulnar shaft with a slight upward curvature of the olecranon, and an elbow joint in which the antebrachium rotated obliquely relative to the humerus. Articular facets on the proximal carpals suggest that the manus of Anisotemnus was habitually extended, indicating a plantigrade stance of the forelimb. Although none of these three taxa have associated hindfoot material, all known Vacan notoungulate astragali have shallow trochlea, well-developed and deep grooves for the flexor hallucis longus, which are separated from the trochlea by a fossa that contains a superior astragalar foramen. An isolated notoungulate pes, not referred to any of the three taxa above, appears to be pentadactyl, having a distinctive, divergent tarsometatarsal joint for its hallux. It also has a shallow trochlea, an astragalar foramen, and a flexor groove, indicating limited rotation of the upper ankle joint. Indeed, a survey of known Casamayoran-aged notoungulate astragali indicates that most taxa had limited mobility at the tibioastragalar joint, in stark contrast to post-Eocene faunas in which nearly all the ungulates had greater rotation of the upper ankle joint and were subcursorial, as evidenced by their longer and deeper trochlear articulation and loss of the astragalar foramen. We suggest that the change from ambulatory- to subcursorial-dominated ungulate faunas across the Eocene-Oliogocene boundary mirrors the changes from brachydont to hypsodont faunas over the same time. Decreased temperatures and rainfall resulting in more open habitats may be related to both morphological evolutionary patterns.