Heterochromatin distribution and chromosomal rearrangements have been proposed as the main sources of karyotype differences among species of Neotropical primates. This variability suggests that there could be differences at other smaller-scale levels of DNA organization as well. In particular, quantitative differences between genomes result from gains and losses of individual DNA segments, and may result in varying genome sizes (C-values) among species. In this work, we studied the genomes of 23 individuals from four species in the genus Ateles (Primates: Platyrrhini): A. chamek, A. paniscus, A. belzebuth, and A. geoffroyi. We analyzed genome size and its relationship with the presence of chromosomal rearrangements and patterns of heterochromatin distribution. The C-value presented in this work for Ateles chamek is the first estimate for this species (3.09 ± 0.23 pg), whereas our estimates for A. belzebuth (2.88 ± 0.06 pg) and A. geoffroyi (3.19 ± 0.24 pg) differed from those previously published. Fluorescent in situ hybridization (FISH) and interspecies comparativegenomic hybridization (iCGH) analyses revealed that differences in genome size among species relate to localized blocks in both heterochromatic and euchromatic regions, the latter of which appear to be genetically unstable. There were also quantitative differences in Y chromosome content. It remains to be seen whether the chromosomal characteristics of Ateles here discussed are common to platyrrhine monkeys, but it is clear that these monkeys exhibit some intriguing genomic features worthy of additional exploration.