This study documents differences in fish assemblages for 32 freshwater streams located between 258 and 2242 m a.s.l. on the eastern slopes of the central range of the Colombian Andes. A total of 2049 fishes belonging to 62 species, 34 genera and 16 families were collected. Species richness declined rapidly with altitude; nearly 90% of the species were recorded between 250 and 1250 m a.s.l. Three of the four physico-chemical variables, of the water, temperature, dissolved oxygen and pH, explained 53.5% of the variation in species richness along the altitudinal gradient, with temperature the most important (37.6%). An analysis of species composition showed that the distinctiveness of the fish fauna increased with elevation, with the greatest turnover observed between 1000 and 1750 m a.s.l. On this altitudinal gradient, turnover was dominated by the loss of species rather than gain, and dominance by just a few species was greater at higher elevations. Turnover was also observed along the altitudinal gradient in the structure of the three functional groups (torrential, pool and pelagic species). The study focused on understanding the pattern of diversity of fish communities inhabiting the Andes in Colombia. Anthropogenic effects on the altitudinal distribution of fish species in the region, however, are largely unknown and would require further investigations.