Because of its close relationship with the process of evolutionary differentiation, it is expected that geographic variability in acoustic sexual traits should be greater among than within populations. This is particularly expected in organisms with typically high population genetic structure and low dispersal abilities, such as anuran amphibians. We studied the acoustic traits of the advertisement call in the small-sized dendrobatoid frog Allobates paleovarzensisthrough its range in Central Amazonia. We accessed the variability of call traits from the within-male to the among-population levels, and evaluated the degree of stereotypy of the call characteristics. Call variability had comparable magnitudes within and among populations, and was independent of the degree of stereotypy of call measurements. Therefore, none of the call traits stood out as a potential cue for discrimination between populations. Spectral call measurements were static and strongly related with body size, which explained between 30 and 35% of the variation of these acoustic traits. Temporal characters of the notes were dynamic and influenced by environmental temperature (e.g., 27% of note rate variation), whilst temporal measurements of the entire calls were not related to the co-factors analysed. Both spectral and temporal call traits varied among populations and between sides of the Amazon River. Our results also indicate that body size and sampling site jointly affected the variability of the call traits. However, geographic distances among populations and the river barrier had no significant effect on the overall acoustic variation, indicating that local stabilising selective forces may be important in the process of call differentiation.