Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Behavior, Animal / drug effects*
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Behavior, Animal / physiology*
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Central Nervous System / drug effects
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Central Nervous System / physiology
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Feeding Behavior / drug effects
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Feeding Behavior / physiology
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Gene Expression / drug effects
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Gene Silencing / drug effects*
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Gene Silencing / physiology
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Gene Targeting / methods
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In Vitro Techniques
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Lymnaea
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Motor Neurons / drug effects
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Motor Neurons / metabolism
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Nitric Oxide / biosynthesis
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Nitric Oxide / physiology*
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Nitric Oxide Synthase / antagonists & inhibitors*
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Nitric Oxide Synthase / genetics
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Nitric Oxide Synthase / metabolism
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Nitric Oxide Synthase Type I
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Phenotype
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RNA, Double-Stranded / pharmacology*
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RNA, Messenger / antagonists & inhibitors
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RNA, Messenger / metabolism
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Reverse Transcriptase Polymerase Chain Reaction
Substances
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RNA, Double-Stranded
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RNA, Messenger
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Nitric Oxide
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Nitric Oxide Synthase
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Nitric Oxide Synthase Type I