Neuropeptide stimulation of the nitric oxide signaling pathway in Drosophila melanogaster Malpighian tubules.
Activation of the nitric oxide (NO) and guanosine 3', 5'-cyclic monophosphate (cGMP) signaling pathway stimulates fluid secretion by the Drosophila melanogaster Malpighian tubule. The neuropeptide cardioacceleratory peptide 2b (CAP2b) has been previously shown to stimulate fluid secretion in this epithelium by elevating intracellular cGMP levels. Therefore, it was of interest to investigate if CAP2b acts through NO in isolated tubules and thus presumably through stimulation of a tubule NO synthase (NOS). We show here by reverse-transcription polymerase chain reaction that Drosophila NOS (dNOS) is expressed in Malpighian tubules. Biochemical assays of NOS activity in whole tubules show that CAP2b significantly stimulates NOS activity. Additionally, fluid secretion and cyclic nucleotide assays show that CAP2b-induced elevation of intracellular cGMP levels and fluid secretion rates are dependent on the activation of a soluble guanylate cyclase. Treatment of tubules with a specific NOS inhibitor abolishes the CAP2b-induced rise in intracellular cGMP levels. These data indicate that CAP2b stimulates NOS and therefore, endogenous NO production, which, in turn, stimulates a soluble guanylate cyclase. This is the first demonstration of stimulation of an endogenous NOS by a defined peptide in Drosophila.
Davies
S A
SA
Division of Molecular Genetics, University of Glasgow, United Kingdom.
Stewart
E J
EJ
Huesmann
G R
GR
Skaer
N J
NJ
Maddrell
S H
SH
Tublitz
N J
NJ
Dow
J A
JA
D000818:Animals; D004331:Drosophila melanogaster / Q000502:physiology*; D006162:Guanylate Cyclase / Q000494:pharmacology; D008317:Malpighian Tubules / Q000187:drug effects* / Q000201:enzymology / Q000502:physiology*; D009479:Neuropeptides / Q000494:pharmacology*; D009569:Nitric Oxide / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D009842:Oligopeptides / Q000494:pharmacology*; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives; D015398:Signal Transduction / Q000187:drug effects*; D012995:Solubility
D009479:Neuropeptides; D009842:Oligopeptides; C095817:cardioacceleratory peptide 2b; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D006162:Guanylate Cyclase; D011761:Pyrrolidonecarboxylic Acid