Amiloride (100 M) was present in the bath during time indicated by solid bars. Oocyte was bathed in Na or K Ringers solution, as indicated by top bars. (A) Current tracing from an oocyte expressing RPK A524V. Diagram and nomenclature are according to Brewster and Bodmer (5).Īctivation of RPK by the Deg mutation. PPK is expressed in one neuron (green) in each cluster of neurons. (J) Schematic diagram of abdominal PNS neurons. (H and I) High magnification views of PPK neurons labeled with anti-PPK antibody showing dendritic arborization and varicosities (I, arrow). Panels show 22C10 staining (E, red), anti-PPK staining (neuron dmd6 F, green), and colocalization (G, yellow). (E and F) High magnification of dorsal cluster of abdominal peripheral neurons double labeled with 22C10 and anti-PPK antibody. Neurons expressing PPK are stained by both antibodies and are yellow (arrows). (D) Abdominal hemisegments labeled with both antiPPK antibody (green) and 22C10 (orange). (C) Peripheral neurons in five abdominal hemisegments labeled with antibody 22C10, which labels peripheral neurons. (A and B) Late stage Drosophila embryos labeled with either an antisense ppk riboprobe (A, purple) or anti-PPK antibody (B, brown). These results suggest that PPK may be a channel subunit involved in mechanosensation.Įmbryonic expression pattern of PPK. In insects, such multiple dendritic neurons play key roles in touch sensation and proprioception and their morphology resembles human mechanosensory free nerve endings. In contrast, PPK was found in sensory dendrites of a subset of peripheral neurons in late stage embryos and early larvae. RPK transcripts were abundant in early stage embryos, suggesting a role in development. Although PPK did not form channels on its own, it associated with and reduced the current generated by a related human brain Na+ channel. Amiloride and gadolinium, which block mechanosensation in vivo, inhibited RPK channels. Expression of RPK generated multimeric Na+ channels that were dominantly activated by a mutation associated with neurodegeneration. We identified two novel Drosophila DEG/ ENaC proteins, Pickpocket (PPK) and Ripped Pocket (RPK). Drosophila melanogaster has proven to be a good model for understanding the physiology of ion channels.
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