The movement of water and small solutes is integral to the survival of freezing and desiccation in insects, yet the underlying mechanisms of these processes are not fully known. Recent evidence suggests that aquaporin (AQP) water channels play critical roles in protecting cells from osmotic damage during freezing and desiccation. Our study sequenced, functionally characterized and measured the tissue abundance of an AQP from freeze-tolerant larvae of the gall fly, Eurosta solidaginis (Diptera Tephritidae). The newly characterized EsAQP1 contains two NPA motifs and six transmembrane regions, and is phylogenetically related to an AQP from the anhydrobiotic chironomid Polypedilum vanderplanki. Using a Xenopus laevis oocyte swelling assay, we demonstrated that EsAQP1 increases water permeability to nine times that of simple diffusion through the membrane. In contrast to its high water permeability, EsAQP1 was impermeable to both glycerol and urea. The abundance of EsAQP1 increased from October to December in all tissues tested and was most abundant in the brain of winter larvae. Because the nervous system is thought to be the primary site of freezing injury, EsAQP1 may cryoprotect the brain from damage associated with water imbalance. The sequence, phylogenetic relationship, osmotic permeability, tissue distribution and seasonal abundance of EsAQP1 further support the role of AQPs in promoting freezing tolerance.