Author(s): Korneev SA, Kemenes I, Straub V, Staras K, Korneeva EI, Kemenes G, Benjamin PR, O'Shea M
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.
Keywords: Animals Behavior, Animal/ drug effects/ physiology Central Nervous System/drug effects/physiology Feeding Behavior/drug effects/physiology Gene Expression/drug effects Gene Silencing/ drug effects/physiology Gene Targeting/methods Lymnaea Motor Neurons/drug effects/metabolism Nitric Oxide/biosynthesis/ physiology Nitric Oxide Synthase/ antagonists & inhibitors/genetics/metabolism Nitric Oxide Synthase Type I Phenotype RNA, Double-Stranded/ pharmacology RNA, Messenger/antagonists & inhibitors/metabolism Reverse Transcriptase Polymerase Chain Reaction