Axonal Localization of transgene mRNA in mature PNS and CNS neurons

Axonal mRNA transport is robust in cultured neurons but there has been limited evidence for this in vivo. We have used a genetic approach to test for in vivo axonal transport of reporter mRNAs. We show that β-actin's 3'-UTR can drive axonal localization of GFP mRNA in mature DRG neurons, b...

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Bibliographic Details
Published in:	The Journal of neuroscience Vol. 31; no. 41; pp. 14481 - 14487
Main Authors:	Willis, Dianna E, Xu, Mei, Donnelly, Christopher J, Tep, Chhavy, Kendall, Marvin, Erenstheyn, Marina, English, Arthur W, Schanen, N Carolyn, Kirn-Safran, Catherine B, Yoon, Sung Ok, Bassell, Gary J, Twiss, Jeffery L
Format:	Journal Article
Language:	English
Published:	United States Society for Neuroscience 12-10-2011
Subjects:	3' Untranslated Regions - genetics Actins - genetics Actins - metabolism Analysis of Variance Animals Axons - metabolism Calcium-Calmodulin-Dependent Protein Kinase Type 2 - genetics Cells, Cultured Dendrites - metabolism Ganglia, Spinal - cytology Gene Expression Regulation - genetics Green Fluorescent Proteins - genetics Mice Mice, Transgenic Microscopy, Confocal - methods Nerve Tissue Proteins - genetics Nerve Tissue Proteins - metabolism Neurons - cytology RNA, Messenger - genetics RNA, Messenger - metabolism RNA, Ribosomal, 18S - genetics RNA, Ribosomal, 18S - metabolism Schwann Cells - metabolism Sciatic Neuropathy - metabolism Sciatic Neuropathy - pathology Spinal Cord - cytology Spinal Cord Injuries - metabolism Spinal Cord Injuries - pathology
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Summary:	Axonal mRNA transport is robust in cultured neurons but there has been limited evidence for this in vivo. We have used a genetic approach to test for in vivo axonal transport of reporter mRNAs. We show that β-actin's 3'-UTR can drive axonal localization of GFP mRNA in mature DRG neurons, but mice with γ-actin's 3'-UTR show no axonal GFP mRNA. Peripheral axotomy triggers transport of the β-actin 3'-UTR containing transgene mRNA into axons. This GFP-3'-β-actin mRNA accumulates in injured PNS axons before activation of the transgene promoter peaks in the DRG. Spinal cord injury also increases axonal GFP signals in mice carrying this transgene without any increase in transgene expression in the DRGs. These data show for the first time that the β-actin 3'-UTR is sufficient for axonal localization in both PNS and CNS neurons in vivo.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Author contributions: N.C.S., C.B.K.-S., A.W.E., S.O.Y., G.J.B., and J.L.T. designed research; D.E.W., M.X., C.J.D., C.T., M.K., and M.E. performed research; A.W.E. and G.J.B. contributed unpublished reagents/analytic tools; D.E.W., C.J.D., N.C.S., C.B.K.-S., S.O.Y., G.J.B., and J.L.T. analyzed data; D.E.W., G.J.B., and J.L.T. wrote the paper. D.E.W., M.X., and C.J.D. share equal contribution to this work.
ISSN:	0270-6474 1529-2401
DOI:	10.1523/JNEUROSCI.2950-11.2011