Chordotonal mechanoreception in insect hearing relies on ciliated sensory neurons that express TRPN and TRPV ion channels. Under high frequency auditory stimulation, these channels may only open for short times, minimizing receptor current. This is counteracted by an elevated electrochemical gradient across the ciliary membrane, achieved by a specialized receptor lymph in the scolopale space enclosed by the scolopale cell. The receptor lymph is thought to be rich in K+ and if it resembles the endolymph of mammalian cochlea, it may also be electro-positive. To test the roles of ion pumps and transporters in generating the receptor lymph in the Drosophila Johnston’s organ, we expressed RNAi constructs that target a panel of ion pump and transporter genes specifically in the scolopale cells, using the nompA-Gal4 driver. We found that subunits of the Na/K-ATPase and the V-ATPase, primary ion pumps, are essential in the scolopale cell for hearing. Furthermore, we found evidence that some secondary transport mechanisms, such as in the cation chloride co-transporter family, which rely on the gradients generated by the primary transport mechanisms, also play a role in this cell. Our results establish a model for receptor lymph generation and help to understand the fundamental mechanisms of mechanotransduction.