title: Subcellular profiling reveals distinct and developmentally regulated repertoire of growth cone mRNAs.
abstract: Cue-directed axon guidance depends partly on local translation in growth cones. Many mRNA transcripts are known to reside in developing axons, yet little is known about their subcellular distribution or, specifically, which transcripts are in growth cones. Here laser capture microdissection (LCM) was used to isolate the growth cones of retinal ganglion cell (RGC) axons of two vertebrate species, mouse and Xenopus, coupled with unbiased genomewide microarray profiling. An unexpectedly large pool of mRNAs defined predominant pathways in protein synthesis, oxidative phosphorylation, cancer, neurological disease, and signaling. Comparative profiling of "young" (pathfinding) versus "old" (target-arriving) Xenopus growth cones revealed that the number and complexity of transcripts increases dramatically with age. Many presynaptic protein mRNAs are present exclusively in old growth cones, suggesting that functionally related sets of mRNAs are targeted to growth cones in a developmentally regulated way. Remarkably, a subset of mRNAs was significantly enriched in the growth cone compared with the axon compartment, indicating that mechanisms exist to localize mRNAs selectively to the growth cone. Furthermore, some receptor transcripts (e.g., EphB4), present exclusively in old growth cones, were equally abundant in young and old cell bodies, indicating that RNA trafficking from the soma is developmentally regulated. Our findings show that the mRNA repertoire in growth cones is regulated dynamically with age and suggest that mRNA localization is tailored to match the functional demands of the growing axon tip as it transforms into the presynaptic terminal.
authors.affiliation: Department of Physiology, Development, and Neuroscience, University of Cambridge, Cambridge CB2 3DY, UK.
authors.forename: Krishna H
authors.initials: KH
authors.lastname: Zivraj
authors.forename: Yi Chun Loraine
authors.initials: YC
authors.lastname: Tung
authors.forename: Michael
authors.initials: M
authors.lastname: Piper
authors.forename: Laura
authors.initials: L
authors.lastname: Gumy
authors.forename: James W
authors.initials: JW
authors.lastname: Fawcett
authors.forename: Giles S H
authors.initials: GS
authors.lastname: Yeo
authors.forename: Christine E
authors.initials: CE
authors.lastname: Holt
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D020869:Gene Expression Profiling / Q000379:methods*; D018507:Gene Expression Regulation, Developmental; D020439:Growth Cones / Q000502:physiology*; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics*; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000502:physiology; D013045:Species Specificity; D013347:Subcellular Fractions / Q000502:physiology; D014982:Xenopus laevis
chemical_list: D012333:RNA, Messenger