Brain size scaling through development in the whitelined sphinx moth (Hyles lineata) shows mass and cell number comparable to flies, bees, and wasps

Factors regulating larval growth and determinants of adult body size are described for several holometabolous insects, but less is known about brain size scaling through development. Here we use the isotropic fractionation ("brain soup") method to estimate the number of brain cells and cell density for the whitelined sphinx moth (Lepidoptera: Hyles lineata) from the first instar through the adult stage. We measure mass and brain cell number and find that, during the larval stages, body mass shows an exponential relationship with head width, while the total number of brain cells increases asymptotically. Larval brain cell number increases by a factor of ten from nearly 8000 in the first instar to over 80,000 in the fifth instar. Brain cell number increases by another factor of 10 during metamorphosis, with the adult brain containing more than 900,000 cells. This is similar to increases during development in the vinegar fly (Drosophila melanogaster) and the black soldier fly (Hermetia illucens). The adult brain falls slightly below the brain-to-body allometry for wasps and bees but is comparable in the number of cells per unit brain mass, indicating a general conservation of brain cell density across these divergent lineages. (c) 2023 Elsevier Ltd. All rights reserved.