Nitric oxide and cyclic GMP induce vesicle release at Drosophila neuromuscular junction.
Nitric oxide (NO) diffuses as short-lived messenger through the plasma membrane and serves, among many other functions, as an activator of the cGMP synthesizing enzyme soluble guanylyl cyclase (sGC). In view of recent genetic investigations that postulated a retrograde signal from the larval muscle fibers to the presynaptic terminals, we looked for the presence of an NO/cGMP signaling system at the neuromuscular junction (NMJ) of Drosophila melanogaster larvae. Application of NO donors induced cGMP immunoreactivity in the presynaptic terminals but not the postsynaptic muscle fibers at an identified NMJ. The NO-induced cGMP immunoreactivity was sensitive to a specific inhibitor (ODQ) of the sGC. Since presynaptic terminals which were surgically isolated from the central nervous system are capable of synthesizing cGMP, we suggest that an NO-sensitive guanylyl cyclase is present in the terminal arborizations. Using a fluorescent dye that is known to stain recycling synaptic vesicles, we demonstrate that NO donors and membrane permeant cGMP analogues cause vesicle release at the NMJ. Moreover, the NO-induced release could be blocked by the specific inhibitor of the sGC. A destaining of synaptic terminals after NO exposure in Ca2+-free solution in the presence of cobalt chloride as a channel blocker suggested that NO stimulates Ca2+-independent vesicle release at the NMJ. The combined immunocytochemical and exocytosis imaging experiments imply the involvement of cGMP and NO in the regulation of vesicle release at the NMJ of Drosophila larvae.
Wildemann
B
B
Institut für Tierökologie und Zellbiologie, Tierärztliche Hochschule Hannover, Germany.
Bicker
G
G
D000818:Animals; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000502:physiology*; D004331:Drosophila melanogaster / Q000254:growth & development / Q000502:physiology*; D006162:Guanylate Cyclase / Q000037:antagonists & inhibitors; D007150:Immunohistochemistry; D007814:Larva; D009469:Neuromuscular Junction / Q000378:metabolism*; D009569:Nitric Oxide / Q000502:physiology*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D010069:Oxadiazoles / Q000494:pharmacology; D017729:Presynaptic Terminals / Q000378:metabolism; D011810:Quinoxalines / Q000494:pharmacology; D013572:Synaptic Vesicles / Q000378:metabolism*
C095284:1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one; D020030:Nitric Oxide Donors; D010069:Oxadiazoles; D011810:Quinoxalines; D009569:Nitric Oxide; D006162:Guanylate Cyclase; D006152:Cyclic GMP