Figure 1

Schematic representation of the experimental design. (A) Male C57BL/6 mice were exposed to drinking water containing 0.015% or 0.03% aspartame for up to 16 weeks. Control groups received plain drinking water over the same duration. Y-maze assay was performed at 4, 8, and 12-weeks during the drinking water exposure. During the 13th week, the mice were bred with females purchased from the vendor and maintained on plain drinking water to produce the F1 generation. Following the breeding, and during weeks 14–15 of the exposures, Barnes maze assay was performed. During the 16th week, tail suspension test (TST) was performed. (B) Beginning at approximately 2-months of age, the F1 male and female mice from each drinking water lineage completed the Y-maze, Barnes maze and TST. Male mice from the 0.03% F1 lineage that did not undergo behavioral testing but were littermates of F1 males that did perform behavioral tests, were bred with females purchased from the vendor and maintained on plain drinking water to produce the F2 generation. Y-maze, and Barnes maze assays (but not TST) were performed in F2 male and female mice beginning at approximately 2 months of age. This Figure was created with BioRender.com.