Imaging Single-mRNA Localization and Translation in Live Neurons

Lee, Byung Hun; Bae, Seong-Woo; Shim, Jae Youn; In, Insik; Park, Hye Yoon

doi:10.14348/molcells.2016.0277

Cited by 18 publications

(15 citation statements)

References 67 publications

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“…Neurons are highly polarized, with long distances between the soma, where RNAs are synthesized in the nucleus, and projections (axons/dendrites/neurites), where cells must quickly respond to stimuli (Figure 1A). These cell types have therefore been a critical model system to understand how RNA localization controls gene expression using imaging‐based approaches as reviewed previously 1–5 . Several neurological diseases, including amyotrophic lateral sclerosis (ALS) and fragile X syndrome (FXS), have been associated with misregulated RNA localization in neurons.…”

Section: Introductionmentioning

confidence: 99%

Mechanisms and consequences of subcellularRNAlocalization across diverse cell types

Engel

Arora

Goering

et al. 2020

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Essentially all cells contain a variety of spatially restricted regions that are important for carrying out specialized functions. Often, these regions contain specialized transcriptomes that facilitate these functions by providing transcripts for localized translation. These transcripts play a functional role in maintaining cell physiology by enabling a quick response to changes in the cellular environment. Here, we review how RNA molecules are trafficked within cells, with a focus on the subcellular locations to which they are trafficked, mechanisms that regulate their transport and clinical disorders associated with misregulation of the process. K E Y W O R D SRNA binding protein, RNA cis-element, RNA localization, RNA transport, zipcode

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Section: Introductionmentioning

confidence: 99%

Mechanisms and consequences of subcellularRNAlocalization across diverse cell types

Engel

Arora

Goering

et al. 2020

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“…Ribo-BiFC technique should provide a method to visualize changes in the subcellular 21 distribution of ribosomes during different stages of Drosophila development and 22 physiological states that will be technically more straightforward than other recently 23 developed methods (Lee et al, 2016). We detected a correlation between the presence of 24 assembled ribosomes and puromycin incorporation, but some of the ribosomes in distal 25 regions of axons seemed not to incorporate puromycin.…”

Section: Ribosomes In the Distal Regions Of Photoreceptor Axons Incormentioning

confidence: 86%

Visualisation of ribosomes in Drosophila axons using Ribo-BiFC

Singh

Abdullahi

Soller

et al. 2019

Preprint

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8Rates of protein synthesis and the number of translating ribosomes vary greatly between 9 different cells in various cell states. The distribution of assembled, and potentially 10 translating, ribosomes within cells can be visualised in Drosophila by using Bimolecular 11Fluorescence Complementation (BiFC) to monitor the interaction between tagged pairs of 12 40S and 60S ribosomal proteins (RPs) that are close neighbours across inter-subunit 13 junctions in the assembled 80S ribosome. Here we describe transgenes that express two 14 novel RP pairs tagged with Venus-based BiFC fragments that considerably increase the 15 sensitivity of this technique that we termed Ribo-BiFC. This improved method should 16 provide a convenient way of monitoring the local distribution of ribosomes in most 17Drosophila cells and we suggest that could be implemented in other organisms. We 18 visualized 80S ribosomes in larval photoreceptors and in other neurons. Assembled 19 ribosomes are most abundant in the various neuronal cell bodies, but they are also present 20 along the lengths of axons and are concentrated in growth cones of larval and pupal 21 photoreceptors. Surprisingly, there is relatively less puromycin incorporation in the distal 22 portion of axons in the optic stalk, suggesting that some of the ribosomes that have 23 started translation may not be engaged in elongation in axons that are still growing. 24Running title: Ribosomes in axon growth cones 25

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“…21,22 However, mRNAs detected by localized tagging might be translated, but in a manner that escapes detection in profiling, which requires ribosomes with their exit tunnels near the membrane 21,22 . Underlying mechanisms may be identified through use of multiple tagging devices in the same cell, live imaging, 8,[18][19][20]90 or strategies that track localization and translation simultaneously 91,92 .…”

Section: Discussionmentioning

confidence: 99%

Records of RNA localization through covalent tagging

Medina-Munoz

Lapointe

Porter

et al. 2019

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RNA movements and localization pervade biology, from embryonic development to disease. To identify RNAs at specific subcellular locations, we anchored a uridine-adding enzyme at those sites, which then marked RNAs in its vicinity with 3' terminal uridines. RNAs were tagged independent of their translation status, and included not only mRNAs, but also ncRNAs and ncRNA processing intermediates. A battery of RNAs, including the stress sensor, IRE1, were tagged at both ER and mitochondria, and reveal RNAs whose dual localization is conserved from yeast to human cells.

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Imaging Single-mRNA Localization and Translation in Live Neurons

Cited by 18 publications

References 67 publications

Mechanisms and consequences of subcellularRNAlocalization across diverse cell types

Mechanisms and consequences of subcellularRNAlocalization across diverse cell types

Visualisation of ribosomes in Drosophila axons using Ribo-BiFC

Records of RNA localization through covalent tagging

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