Abstract Anolis lizards have been a model of study in ecomorphology in the Caribbean islands because species with the same type of microhabitat share similar morphological features. But despite their great diversity, little is known about continental species. We analyzed the relationship between the anatomical characteristics of the appendicular skeleton and the locomotor performance of two Anolis species found in Colombia that have different use of habitat. Anolis heterodermus , a strictly arboreal species, was compared with Anolis tolimensis , which inhabits the lower strata of vegetation. These two species differ in their body plan not only in body shape and external morphological features, but also in the skeleton and appendicular musculature. The results highlight the muscle and bone specializations associated with the use of habitat in this genus, such as the presence of more robust bones to enlarge the surface of muscle insertion, the thickening and loss of carpal parts, thickening of tendons associated with the manus, and greater development of muscle mass in the forelimbs by A. heterodermus with respect to A. tolimensis . These differences are related to the use of the microhabitat and the locomotor style of each species.