The Drosophila melanogaster homologue of an insect calcitonin-like diuretic hormone was identified in a BLAST search of the Drosophila genome database. The predicted 31-residue amidated peptide (D. melanogaster DH(31); Drome-DH(31)) was synthesised and tested for activity on fruit fly Malpighian tubules. It increases tubule secretion by approximately 35 % of the response obtained with a myokinin from the housefly Musca domestica (muscakinin; Musdo-K) and has an EC(50) of 4.3 nmol l(−)(1). The diuretic activities of Drome-DH(31) and Musdo-K were additive when tested at threshold and supra-maximal concentrations, which suggests that they target different transport processes. In support of this, Drome-DH(31) increased the rate of secretion by tubules held in bathing fluid with a reduced Cl(−) concentration, whereas Musdo-K did so only in the presence of Drome-DH(31). Stimulation with Drome-DH(31) increased the lumen-positive transepithelial potential in the main secretory segment of the tubule. This was attributed to activation of an apical electrogenic proton-translocating V-ATPase in principal cells, since it was associated with hyperpolarisation of the apical membrane potential and acidification of secreted urine by 0.25 pH units. Exogenous 8-bromo-cyclic AMP and cyclic GMP increased tubule secretion to the same extent as Drome-DH(31) and, when tested together with the diuretic peptide, their activities were not additive. Stimulation with Drome-DH(31) resulted in a dose-dependent increase in cyclic AMP production by tubules incubated in saline containing 0.5 mmol l(−)(1) 3-isobutyl-1-methylxanthine, whereas cyclic GMP production was unchanged. Taken together, the data are consistent with Drome-DH(31) activating an apical membrane V-ATPase via cyclic AMP. Since the K(+) concentration of the secreted urine was unchanged, it is likely that Drome-DH(31) has an equal effect on K(+) and Na(+) entry across the basolateral membrane.
The Drosophila melanogaster homologue of an insect calcitonin-like diuretic peptide stimulates V-ATPase activity in fruit fly Malpighian tubules
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G.M. Coast, S.G. Webster, K.M. Schegg, S.S. Tobe, D.A. Schooley; The Drosophila melanogaster homologue of an insect calcitonin-like diuretic peptide stimulates V-ATPase activity in fruit fly Malpighian tubules. J Exp Biol 15 May 2001; 204 (10): 1795–1804. doi: https://doi.org/10.1242/jeb.204.10.1795
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