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RNAi pathway is functional in peripheral nerve axons

Alexander K. Murashov^*,1, Vishnu Chintalgattu^*,2, Rustem R. Islamov^#,2, Teresa E. Lever^,2, Elena S. Pak^*, Paulina L. Sierpinski^*, Laxmansa C. Katwa^* and Michael R. Van Scott^*

^* Department of Physiology, and

Department of Communication Sciences and Disorders, East Carolina University, Greenville, North Carolina, USA; and

^# Department of Histology, Kazan State Medical University, Kazan, Russia

¹Correspondence: Department of Physiology, Brody School of Medicine, East Carolina University, 600 Moye Blvd., Greenville, NC 27834, USA. E-mail: murashoval{at}ecu.edu

Recent observations demonstrated that translation of mRNAs may occur in axonal processes at sites that are long distances away from the neuronal perikaria. While axonal protein synthesis has been documented in several studies, the mechanism of its regulation remains unclear. The aim of this study was to investigate whether RNA interference (RNAi) may be one of the pathways that control local protein synthesis in axons. Here we show that sciatic nerve contains Argonaute2 nuclease, fragile X mental retardation protein, p100 nuclease, and Gemin3 helicase—components of the RNA-induced silencing complex (RISC). Application of short-interfering RNAs against neuronal ß-tubulin to the sciatic nerve initiated RISC formation, causing a decrease in levels of neuronal ß-tubulin III mRNA and corresponding protein, as well as a significant reduction in retrograde labeling of lumbar motor neurons. Our observations indicate that RNAi is functional in peripheral mammalian axons and is independent from the neuronal cell body or Schwann cells. We introduce a concept of local regulation of axonal translation via RNAi.—Murashov, A. K., Chintalgattu, V., Islamov, R. R., Lever, T. E., Pak, E. S., Sierpinski, P. L., Katwa, L. C., Van Scott, M. R. RNAi pathway is functional in peripheral nerve axons.

Key Words: RNA interference • axonal protein synthesis • siRNA • sciatic nerve • DRG

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