title: Analysis of piRNA-Like Small Non-coding RNAs Present in Axons of Adult Sensory Neurons.
abstract: Small non-coding RNAs (sncRNAs) have been shown to play pivotal roles in spatiotemporal-specific gene regulation that is linked to many different biological functions. PIWI-interacting RNAs (piRNAs), typically 25-34-nucleotide long, are originally identified and thought to be restricted in germline cells. However, recent studies suggest that piRNAs associate with neuronal PIWI proteins, contributing to neuronal development and function. Here, we identify a cohort of piRNA-like sncRNAs (piLRNAs) in rat sciatic nerve axoplasm and directly contrast temporal changes of piLRNA levels in the nerve following injury, as compared with those in an uninjured nerve using deep sequencing. We find that 32 of a total of 53 annotated piLRNAs show significant changes in their levels in the regenerating nerve, suggesting that individual axonal piLRNAs may play important regulatory roles in local messenger RNA (mRNA) translation during regeneration. Bioinformatics and biochemical analyses show that these piLRNAs carry characteristic features of mammalian piRNAs, including sizes, a sequence bias for uracil at the 5'-end and a 2'-O-methylation at the 3'-end. Their axonal expression is directly visualized by fluorescence in situ hybridization in cultured dorsal root ganglion neurons as well as immunoprecipitation with MIWI. Further, depletion of MIWI protein using RNAi from cultured sensory neurons increases axon growth rates, decreases axon retraction after injury, and increases axon regrowth after injury. All these data suggest more general roles for MIWI/piLRNA pathway that could confer a unique advantage for coordinately altering the population of proteins generated in growth cones and axons of neurons by targeting mRNA cohorts.
authors.affiliation: Nemours Biomedical Research, Alfred I duPont Hospital for Children, Wilmington, DE, 19803, USA.|Department of Biological Sciences, University of Delaware, Newark, DE, 19716, USA.
authors.forename: Monichan
authors.initials: M
authors.lastname: Phay
authors.affiliation: Nemours Biomedical Research, Alfred I duPont Hospital for Children, Wilmington, DE, 19803, USA.
authors.forename: Hak Hee
authors.initials: HH
authors.lastname: Kim
authors.affiliation: Nemours Biomedical Research, Alfred I duPont Hospital for Children, Wilmington, DE, 19803, USA. yoo@medsci.udel.edu.|Department of Biological Sciences, University of Delaware, Newark, DE, 19716, USA. yoo@medsci.udel.edu.
authors.forename: Soonmoon
authors.identifier: 0000-0002-1645-6809
authors.initials: S
authors.lastname: Yoo
mesh_terms: D000375:Aging / Q000378:metabolism*; D000818:Animals; D060565:Argonaute Proteins / Q000378:metabolism; D001369:Axons / Q000378:metabolism*; D005727:Ganglia, Spinal / Q000378:metabolism; D005786:Gene Expression Regulation; D008297:Male; D034741:RNA, Small Interfering / Q000378:metabolism*; D058727:RNA, Small Untranslated / Q000235:genetics / Q000378:metabolism*; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012584:Sciatic Nerve / Q000293:injuries / Q000378:metabolism; D011984:Sensory Receptor Cells / Q000378:metabolism*; D013997:Time Factors
chemical_list: D060565:Argonaute Proteins; D034741:RNA, Small Interfering; D058727:RNA, Small Untranslated
keywords: Axon growth; Intra-axonal translation; Neuronal piRNA; Regenerating nerve; Small RNA sequencing; Small non-coding RNA