期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 115, 期 41, 页码 E9697-E9706出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1806189115
关键词
RNA localization; axon; beta-actin; molecular beacon; single molecule
资金
- Wellcome Trust [102457/Z/13/A, 085314/Z/08/Z]
- European Research Council [322817]
- EPSRC [EP/R025398/1] Funding Source: UKRI
- Wellcome Trust [102457/Z/13/A] Funding Source: Wellcome Trust
During embryonic nervous system assembly, mRNA localization is precisely regulated in growing axons, affording subcellular autonomy by allowing controlled protein expression in space and time. Different sets of mRNAs exhibit different localization patterns across the axon. However, little is known about how mRNAs move in axons or how these patterns are generated. Here, we couple molecular beacon technology with highly inclined and laminated optical sheet microscopy to image single molecules of identified endogenous mRNA in growing axons. By combining quantitative single-molecule imaging with biophysical motion models, we show that beta-actin mRNA travels mainly as single copies and exhibits different motion-type frequencies in different axonal subcompartments. We find that beta-actin mRNA density is fourfold enriched in the growth cone central domain compared with the axon shaft and that a modicum of directed transport is vital for delivery of mRNA to the axon tip. Through mathematical modeling we further demonstrate that directional differences in motor-driven mRNA transport speeds are sufficient to generate beta-actin mRNA enrichment at the growth cone. Our results provide insight into how mRNAs are trafficked in axons and a mechanism for generating different mRNA densities across axonal subcompartments.
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