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We reconstruct the historical biogeography of cichlid fishes endemic to the trans-Andean region of NW South America. DNA sequences were used to study historical biogeography of the cichlid genera Andinoacara (Cichlasomatini) and Mesoheros (Heroini). Two eventbased methodological approaches, parsimony-based Statistical Dispersal-Vicariance Analysis (S-DIVA) and likelihood-based Dispersal-Extinction Cladogenesis (DEC in Lagrange) were used for ancestral-area reconstructions. Molecular clock analysis of the whole group of Neotropical Cichlidae (using mtDNA and nucDNA markers) was calibrated using BEAST by six known cichlid fossils. The historical biogeography of both studied trans-Andean cichlid genera is best explained by a series of vicariance events that fragmented an ancestrally wider distribution. Both genera have a highly congruent vicariant historical biogeography in their shared distribution in the Colombian-Ecuadorian Choco. The Andean uplift and formation of the Central American isthmus strongly impacted the distribution patterns of the freshwater ichtyofauna in the NW Neotropics as suggested by the historical biogeography of the two studied cichlid groups. Despite strong congruence in their historical biogeography the two studied cichlid lineages (part of the tribe Cichlasomatini and Heroini, respectively) have highly different evolutionary substitution rates in the studied mtDNA cytb marker.