Abstract Aim We analysed the distributional patterns of six terrestrial vertebrate taxa from the Oaxacan Highlands (Sierra Mazateca, Nudo de Zempoaltépetl and Sierra de Juárez) through a cladistic biogeographical approach, in order to test their naturalness as a biotic unit. Location The Oaxacan Highlands, Mexico. Methods The cladistic biogeographical analysis was based on the area cladograms of the Pseudoeurycea bellii species group (Amphibia: Plethodontidae), the genus Chlorospingus (Aves: Thraupidae), the genera Microtus, Reithrodontomys and Habromys, and the Peromyscus aztecus species group (Mammalia: Rodentia). We obtained paralogy-free subtrees, from which the components were coded in a data matrix for parsimony analysis. The data matrix was analysed with Nonathrough WinClada. Results The parsimony analysis resulted in a single general area cladogram in which areas were fragmented following the sequence Sierra Madre Occidental, Trans-Mexican Volcanic Belt, Chiapas, Sierra Madre Oriental + Sierra Mazateca, Sierra Madre del Sur, Nudo de Zempoaltépetl and Sierra de Juárez. Main conclusions The general area cladogram shows that the Oaxacan Highlands do not constitute a natural unit. The Sierra Mazateca is the sister area to the Sierra Madre Oriental, whereas the Nudo de Zempoaltépetl and the Sierra de Juárez are closely related to the Sierra Madre del Sur. The events that might have caused these patterns include cycles of expansion and contraction of mountain pinyon, juniper and oak woodlands during the Pleistocene.