title: Messenger RNAs localized to distal projections of human stem cell derived neurons.
abstract: The identification of mRNAs in distal projections of model organisms has led to the discovery of multiple proteins that are locally synthesized for functional roles such as axon guidance, injury signaling and regeneration. The extent to which local protein synthesis is conserved in human neurons is unknown. Here we used compartmentalized microfluidic chambers to characterize the transcriptome of distal projections of human embryonic stem cells differentiated using a protocol which enriched for glutamatergic neurons (hESC-neurons). Using gene expression analysis, we identified mRNAs proportionally enriched in these projections, representing a functionally unique local transcriptome as compared to the human neuronal transcriptome inclusive of somata. Further, we found that the most abundant mRNAs within these hESC-neuron projections were functionally similar to the axonal transcriptome of rat cortical neurons. We confirmed the presence of two well characterized axonal mRNAs in model organisms, β-actin and GAP43, within hESC-neuron projections using multiplexed single molecule RNA-FISH. Additionally, we report the novel finding that oxytocin mRNA localized to these human projections and confirmed its localization using RNA-FISH. This new evaluation of mRNA within human projections provides an important resource for studying local mRNA translation and has the potential to reveal both conserved and unique translation dependent mechanisms.
authors.affiliation: Curriculum in Genetics and Molecular Biology, UNC-Chapel Hill, Chapel Hill, NC, 27599, USA.|UNC/NC State Joint Department of Biomedical Engineering, UNC-Chapel Hill, Chapel Hill, NC, 27599, USA.
authors.forename: Rebecca L
authors.identifier: http://orcid.org/0000-0002-0075-6188
authors.initials: RL
authors.lastname: Bigler
authors.affiliation: UNC/NC State Joint Department of Biomedical Engineering, UNC-Chapel Hill, Chapel Hill, NC, 27599, USA.
authors.forename: Joyce W
authors.initials: JW
authors.lastname: Kamande
authors.affiliation: UNC Human Pluripotent Stem Cell Core Facility, UNC-Chapel Hill, Chapel Hill, NC, 24599, USA.
authors.forename: Raluca
authors.initials: R
authors.lastname: Dumitru
authors.affiliation: UNC/NC State Joint Department of Biomedical Engineering, UNC-Chapel Hill, Chapel Hill, NC, 27599, USA.|Neuroscience Center, UNC-Chapel Hill, Chapel Hill, NC, 27599, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Niedringhaus
authors.affiliation: UNC/NC State Joint Department of Biomedical Engineering, UNC-Chapel Hill, Chapel Hill, NC, 27599, USA. anne.marion.taylor@gmail.com.|Neuroscience Center, UNC-Chapel Hill, Chapel Hill, NC, 27599, USA. anne.marion.taylor@gmail.com.|Carolina Institute for Developmental Disabilities, Chapel Hill, NC, 27599, USA. anne.marion.taylor@gmail.com.
authors.forename: Anne Marion
authors.identifier: http://orcid.org/0000-0003-0998-087X
authors.initials: AM
authors.lastname: Taylor
mesh_terms: D001369:Axons / Q000378:metabolism*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D019295:Computational Biology / Q000379:methods; D053595:Embryonic Stem Cells / Q000166:cytology; D063990:Gene Ontology; D006801:Humans; D017403:In Situ Hybridization; D046210:Microfluidic Analytical Techniques; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D034443:RNA Transport; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology*; D059467:Transcriptome
chemical_list: D012333:RNA, Messenger