Supplementary material from "Allometric analysis of brain cell number in hymenoptera suggests ant brains diverge from general trends"

Many comparative neurobiological studies seek to connect sensory or behavioural attributes across taxa with differences in their brain composition. Recent studies in vertebrates suggest cell number and density may be better correlated with behavioural ability than brain mass or volume, but few estimates of such figures exist for insects. Here, we use the isotropic fractionator method to estimate total brain cell numbers for 32 species of Hymenoptera spanning seven subfamilies. We find estimates from using this method are comparable to traditional, whole-brain cell counts of two species and to published estimates from established stereological methods. We present allometric scaling relationships between body and brain mass, brain mass and nuclei number, and body mass and cell density and find that ants stand out from bees and wasps as having particularly small brains by measures of mass and cell number. We find that Hymenoptera follow the general trend of smaller animals having proportionally larger brains. Smaller Hymenoptera also feature higher brain cell densities than the larger ones, as is the case in most vertebrates, but in contrast with primates, in which neuron density remains rather constant across changes in brain mass. Overall, our findings establish the isotropic fractionator as a useful method for comparative studies of brain size evolution in insects.