Cold tolerance of Drosophila species is tightly linked to epithelial K⁺ transport capacity of the Malpighian tubules and rectal pads

Insect chill tolerance is strongly associated with the ability to maintain ion and water homeostasis during cold exposure. Maintenance of K⁺ balance is particularly important due to its role in setting the cell membrane potential that is involved in many aspects of cellular function and viability. In most insects, K⁺ balance is maintained through secretion at the Malpighian tubules balancing reabsorption from the hindgut and passive leak arising from the gut lumen. Here, we used a scanning ion-selective electrode technique (SIET) system at benign (23°C) and low (6°C) temperature to examine K⁺ flux across the Malpighian tubules and the rectal pads in the hindgut in five Drosophila species that differ in cold tolerance. We found that chill tolerant species were better at maintaining K⁺ secretion and supressing reabsorption during cold exposure. In contrast, chill susceptible species exhibited large reductions in secretion with no change, or a paradoxical increase, in K⁺ reabsorption. Using an assay to measure paracellular leak we found that chill susceptible species experience a large increase in leak during cold exposure, which could explain the increased K⁺ reabsorption found in these species. Our data therefore strongly support the hypothesis that cold tolerant Drosophila species are better at maintaining K⁺ homeostasis through an increased ability to maintain K⁺ secretion rates and through reduced leakage of K⁺ towards the hemolymph. These adaptations are manifested both at the Malpighian tubule and at the rectal pads in the hindgut and ensure that cold tolerant species experience less perturbation of K⁺ homeostasis during cold stress.