title: Plasmids containing NRSE/RE1 sites enhance neurite outgrowth of retinal ganglion cells via sequestration of REST independent of NRSE dsRNA expression.
abstract: Repressor element-1 silencing transcription factor (REST) is a transcriptional repressor of neuron-specific genes that binds to a conserved DNA element, the neuron restrictive silencer element (NRSE/RE1). Interestingly, increased REST activity is found in several neurological diseases like Huntington's disease and cerebral ischemia. Recently, it was shown that NRSE dsRNA, a double-stranded non-coding RNA can bind to REST during a defined period of neuronal differentiation, and thereby changes REST from a transcriptional repressor to an activator of neuron-specific genes. Here, we analyzed the effects of NRSE dsRNA expression in primary retinal ganglion cells. We found that NRSE dsRNA expression vectors significantly enhance neurite outgrowth even when axonal degeneration is induced by neurotrophin deprivation. Transfection of HEK cells with NRSE dsRNA-expressing vectors altered their morphology leading to the formation of thin processes and induced the expression of neurofilament-68. Surprisingly, control vectors containing REST-binding sites, but not expressing NRSE dsRNA, resulted in the same effects, also in the retinal ganglion cell model. Reporter assays and retention of REST in the cytoplasm with a labeled NRSE/RE1-containing plasmid incapable of entering the nucleus suggest that sequestration of REST in the cytoplasm is the reason for the observed effects. No evidence for a biological function of NRSE dsRNA could be found in these models. We conclude that sequestration of REST leads to enhanced neurite outgrowth in retinal ganglion cells and that an increased activity of REST, as it is found in several neurodegenerative diseases, can be effectively modulated by sequestration of REST with plasmids containing NRSE/RE1 sites.
authors.affiliation: Department of Neurology, University Medicine Göttingen, Göttingen, Germany. jkoch@med.uni-goettingen.de
authors.forename: Jan C
authors.initials: JC
authors.lastname: Koch
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Barski
authors.forename: Paul
authors.initials: P
authors.lastname: Lingor
authors.forename: Mathias
authors.initials: M
authors.lastname: Bähr
authors.forename: Uwe
authors.initials: U
authors.lastname: Michel
mesh_terms: D000818:Animals; D001483:Base Sequence; D002470:Cell Survival; D002478:Cells, Cultured; D016384:Consensus Sequence; D003593:Cytoplasm / Q000378:metabolism; D004261:DNA Replication; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism* / Q000627:therapeutic use; D057809:HEK293 Cells; D006801:Humans; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D016501:Neurites / Q000378:metabolism* / Q000473:pathology; D019636:Neurodegenerative Diseases / Q000628:therapy; D016900:Neurofilament Proteins / Q000378:metabolism; D034741:RNA, Small Interfering / Q000737:chemistry / Q000378:metabolism / Q000627:therapeutic use; D051381:Rats; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D012165:Retinal Ganglion Cells / Q000378:metabolism* / Q000473:pathology; D037941:Silencer Elements, Transcriptional; D000099229:RE1-Silencing Transcription Factor
chemical_list: D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D034741:RNA, Small Interfering; D012097:Repressor Proteins; D000099229:RE1-Silencing Transcription Factor; C066858:neurofilament protein NF 68