The present study involved an electrophoretic survey of 22 protein loci in 269 individuals belonging to three species of the genus Akodon, A. aff. cursor (2n = 16), A. cursor (2n = 14/15), and A. montensis (2n = 24/25/26), collected in Eastern Brazil. The joint results of gene diversity, genetic distances, phenetic analyses, and phylogenetic trees suggested that A. aff. cursor has recently separated from A. cursor and that the three species have experienced a recent chromosomal divergence followed by low allozyme differentiation. These data are in agreement with their classification as sibling species.