Vertebrates exhibit diversity in the presence and number of aquaporin (Aqp)-10 genes. In Rodentia, mice possess an Aqp10 pseudogene, whereas guinea pigs possess an intact Aqp10. However, Aqp10 retention and pseudogenization history in various rodent lineages remains unclear. Therefore, in this study, we aimed to investigate the molecular evolution of Aqp10 using the recent increasingly decoded rodent genome sequences. We analyzed Aqp10 in the genomes of 43 rodent species belonging to 14 families and found that Aqp10 was pseudogenized in 13 species of three families in the Myomorpha suborder. In contrast, a single intact Aqp10 was retained in the other 30 rodent species, with no Aqp10 pseudogene found in the Castorimorpha, Hystricomorpha, and Sciuromorpha suborders. Additionally, we investigated the tissue expression levels of aquaglyceroporin genes in guinea pigs and rats via reverse transcription-polymerase chain reaction and detected Aqp10 expression in the guinea pig intestines. Notably, none of the examined rat organs expressed Aqp10
however, Aqp7 was expressed in the rat intestines. In situ hybridization showed that guinea pig Aqp10 was expressed in the intestinal epithelial cells. Moreover, AQP10 was permeable to water, glycerol, urea, and boric acid in Xenopus oocytes. Overall, these results clarify the Aqp10 pseudogenization history in Rodentia and suggest guinea pigs as excellent small animal models to analyze the intestinal AQP10 functions.