Suppression of nitric oxide (NO)-dependent behavior by double-stranded RNA-mediated silencing of a neuronal NO synthase gene.
We have used double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) to disrupt neuronal nitric oxide (NO) synthase (nNOS) gene function in the snail Lymnaea stagnalis and have detected a specific behavioral phenotype. The injection of whole animals with synthetic dsRNA molecules targeted to the nNOS-encoding mRNA reduces feeding behavior in vivo and fictive feeding in vitro and interferes with NO synthesis by the CNS. By showing that synthetic dsRNA targeted to the nNOS mRNA causes a significant and long-lasting reduction in the levels of Lym-nNOS mRNA, we verify that specific RNAi has occurred. Importantly, our results establish that the expression of nNOS gene is essential for normal feeding behavior. They also show that dsRNA can be used in the investigation of functional gene expression in the context of whole animal behavior, regardless of the availability of targeted mutation technologies.
Korneev
Sergei A
SA
Sussex Centre for Neuroscience, University of Sussex, Brighton BN1 9QG, United Kingdom.
Kemenes
Ildiko
I
Straub
Volko
V
Staras
Kevin
K
Korneeva
Elena I
EI
Kemenes
Gyorgy
G
Benjamin
Paul R
PR
O'Shea
Michael
M
D000818:Animals; D001522:Behavior, Animal / Q000187:drug effects* / Q000502:physiology*; D002490:Central Nervous System / Q000187:drug effects / Q000502:physiology; D005247:Feeding Behavior / Q000187:drug effects / Q000502:physiology; D015870:Gene Expression / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects* / Q000502:physiology; D018390:Gene Targeting / Q000379:methods; D066298:In Vitro Techniques; D008195:Lymnaea; D009046:Motor Neurons / Q000187:drug effects / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D052248:Nitric Oxide Synthase Type I; D010641:Phenotype; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I