Menu

Close

mRNA localisation in endothelial cells regulates blood vessel sprouting

Guilherme Costa, Nawseen Tarannum, Shane Herbert

Preprint posted on August 06, 2018 https://www.biorxiv.org/content/early/2018/08/06/374850

rab13 transcripts are targeted to and locally translated at the leading edge of migrating endothelial cells which fine-tunes their protrusive activity during developmental angiogenesis.

Selected by Andreas van Impel

Background

The establishment of polarity on the cellular level by asymmetrically localizing cellular components and molecules including mRNAs plays a central role during various biological processes. For example, the localization of maternal bicoid, oskar and gurken mRNA to opposing ends within the Drosophila oocyte is key for early axis formation in the embryo. Also during asymmetric cell divisions certain proteins and mRNAs are targeted to specific subcellular territories and hence end up in only one of the daughter cells, thereby triggering a distinct developmental program in each of the cells [1].

During cell migration, a polar distribution of proteins regulating the cytoskeleton has been shown to be important for establishing and maintaining directionality. In this context, the accumulation of certain mRNAs to protrusions of different migratory cell types has been reported and it is thought that their local translation contributes to the regulation of actin and focal adhesion dynamics at the leading edge [2]. However, to what extent such dynamic subcellular distributions of certain transcripts are involved in more complex morphogenetic events such as sprouting angiogenesis (a process that generates new blood vessels by sprouting and migration of endothelial cells from pre-existing vessels) has not been explored.

 

Key findings

In this preprint, Costa and colleagues investigate the presence and functional relevance of asymmetrically located transcripts in migrating endothelial cells. In order to identify such transcripts they employ Human Umbilical Vein Endothelial cells (HUVECs) in a modified Boyden chamber, which allows the separation of cell protrusions from the cell bodies. Subsequent RNA-Seq analysis of these samples revealed a set of 320 mRNAs that are enriched in the protrusions of HUVECs, many of which encode for proteins that are known to accumulate at the leading edge of migrating cells and that are involved in the regulation of cell motility. One of the strongest enrichments within the cell protrusion fraction was detected for the mRNA of the small GTPase RAB13, a known regulator of cellular vesicle trafficking that has been previously reported to be required for directional migration during zebrafish angiogenesis [3]. The authors subsequently show that RAB13 mRNA indeed localizes to the distal most region of endothelial tip cells in vitro and that the transcripts are also actively translated at the leading edge of endothelial cells. By deleting different parts of the RAB13 3’UTR Costa et al. identify a specific 192nt ‘zipcode’ sequence that is essential and sufficient to target mRNAs to the leading edge of migrating endothelial cells. To test these findings also in a more complex in vivo situation, the authors establish a vascular specific in vivo RNA labelling system in zebrafish embryos and show that also during embryonic angiogenic sprouting the rab13 3’UTR sequence is capable of localizing transcripts to the leading edge of endothelial cells.
Finally, to assess a functional role for the specific subcellular localization of rab13 mRNA during angiogenesis, the authors generate zebrafish mutants carrying a deletion within the rab13 3’UTR that strongly reduces the polarized appearance of the transcripts (but does not cause a general loss of the mRNA). Subsequent time-lapse analysis during arterial sprouting and formation of intersegmental blood vessels reveals that in the absence of localized rab13 mRNA the sprouting vessels frequently form transient ectopic branches at distinct positions amongst their migration route, likely reflecting problems to find the correct migration path. These observations suggest that the correct subcellular accumulation of rab13 mRNA at the leading edge provides critical positional information that is required for fine tuning of angiogenic sprouting in vivo.

 

Why this is cool

The results from Costa et al clearly indicate that the subcellular targeting of certain transcripts is not only an important feature of migrating fibroblasts, neurons or cancer cells, but that this concept of subcellular asymmetry is also required for proper developmental angiogenesis. It is remarkable, that the disturbance of rab13 transcript localization (and not its overall expression) during angiogenesis is sufficient to affect the migratory behaviour of endothelial cells. Interestingly, the morpholino-mediated knock-down of rab13 in zebrafish was previously shown to impede the formation of intersegmental vessels and to interfere with the directionality of tip cells, suggesting that the here reported defects upon mislocalization of rab13 transcripts represent a hypomorphic situation reflecting the importance of transcript localization and local translation of Rab13 at the leading edge for its migratory function. These findings therefore imply that subcellular targeting of certain transcripts during angiogenesis constitutes another level of complexity during the regulation of endothelial cell behaviour and directional migration.

 

Open questions

  • How is the accumulation of rab13 mRNA at the leading edge mediated, which interaction partners are involved in the transport and anchoring of the transcripts?
  • What is the local signal preceding the dynamic accumulation of transcripts at a certain site?
  • What transcripts other than those encoding cytoskeletal regulators are enriched within protrusions? Are also mRNAs encoding components of certain signalling pathways enriched at the leading edge?

 

 

Further reading

[1] Cody, N.E.A. et al., The many functions of mRNA localization during normal development and disease: from pillar to post. WIREs Dev Biol, 2:781–796. (2013)

[2] Liao, G. et al., Control of cell migration through mRNA localization and local translation. WIREs RNA, 6:1–15. (2015)

[3] Wu, C., et al., Rab13-dependent Trafficking of RhoA Is Required for Directional Migration and Angiogenesis. J Biol Chem, 286(26):23511-20. (2011)

 

 

Posted on: 29th August 2018 , updated on: 31st August 2018

Read preprint (No Ratings Yet)




  • Have your say

    Your email address will not be published. Required fields are marked *

    Sign up to customise the site to your preferences and to receive alerts

    Register here

    Also in the cell biology category:

    SABER enables highly multiplexed and amplified detection of DNA and RNA in cells and tissues

    Jocelyn Y. Kishi, Brian J. Beliveau, Sylvain W. Lapan, et al.



    Selected by Yen-Chung Chen

    PIKfyve/Fab1 is required for efficient V-ATPase and hydrolase delivery to phagosomes, phagosomal killing, and restriction of Legionella infection

    Catherine M Buckley, Victoria L Heath, Aurelie Gueho, et al.



    Selected by Giuliana Clemente

    1

    Memory sequencing reveals heritable single cell gene expression programs associated with distinct cellular behaviors

    Sydney M Shaffer, Benjamin L Emert, Ann E. Sizemore, et al.



    Selected by Leighton Daigh

    1

    Aurora A depletion reveals centrosome-independent polarization mechanism in C. elegans

    Kerstin Klinkert, Nicolas Levernier, Peter Gross, et al.

    AND

    Centrosome Aurora A gradient ensures a single PAR-2 polarity axis by regulating RhoGEF ECT-2 localization in C. elegans embryos

    Sachin Kotak, Sukriti Kapoor



    Selected by Giuliana Clemente

    Anti-angiogenic effects of VEGF stimulation on endothelium deficient in phosphoinositide recycling

    Amber N Stratman, Olivia M Farrelly, Constantinos M Mikelis, et al.



    Selected by Coert Margadant

    Neural crest cells regulate optic cup morphogenesis by promoting extracellular matrix assembly

    Chase Dallas Bryan, Rebecca Lynne Pfeiffer, Bryan William Jones, et al.



    Selected by Ashrifia Adomako-Ankomah

    1

    Local protein synthesis in axon terminals and dendritic spines differentiates plasticity contexts

    Anne-Sophie Hafner, Paul Donlin-Asp, Beulah Leitch, et al.



    Selected by Dipen Rajgor

    The cytoskeleton as a smart composite material: A unified pathway linking microtubules, myosin-II filaments and integrin adhesions

    Nisha Mohd Rafiq, Yukako Nishimura, Sergey V. Plotnikov, et al.



    Selected by Coert Margadant

    Quantitative, real-time, single cell analysis in tissue reveals expression dynamics of neurogenesis

    Cerys S Manning, Veronica Biga, James Boyd, et al.



    Selected by Teresa Rayon

    Profiling the surface proteome identifies actionable biology for TSC1 mutant cells beyond mTORC1 signaling

    Junnian Wei, Kevin K. Leung, Charles Truillet, et al.



    Selected by Rob Hynds

    1

    Optogenetic dissection of mitotic spindle positioning in vivo

    Lars-Eric Fielmich, Ruben Schmidt, Daniel J Dickinson, et al.



    Selected by Angika Basant

    1

    Excitable RhoA dynamics drive pulsed contractions in the early C. elegans embryo.

    Jonathan B Michaux, Francois B Robin, William M McFadden, et al.



    Selected by Sundar Naganathan

    Moving beyond P values: Everyday data analysis with estimation plots

    Joses Ho, Tayfun Tumkaya, Sameer Aryal, et al.



    Selected by Gautam Dey

    1

    A limited number of double-strand DNA breaks are sufficient to delay cell cycle progression.

    Jeroen van den Berg, Anna G. Manjon, Karoline Kielbassa, et al.



    Selected by Leighton Daigh

    Optogenetic manipulation of medullary neurons in the locust optic lobe

    Hongxia Wang, Richard B. Dewell, Markus U. Ehrengruber, et al.



    Selected by Ana Patricia Ramos

    JNK-mediated spindle reorientation in stem cells promotes dysplasia in the aging intestine

    Daniel Hu, Heinrich Jasper



    Selected by Maiko Kitaoka

    Also in the developmental biology category:

    Polyacrylamide Bead Sensors for in vivo Quantification of Cell-Scale Stress in Zebrafish Development

    Nicole Traeber, Klemens Uhlmann, Salvatore Girardo, et al.



    Selected by Jacky G. Goetz

    millepattes micropeptides are an ancient developmental switch required for embryonic patterning

    Suparna Ray, Miriam I Rosenberg, Hélène Chanut-Delalande, et al.



    Selected by Erik Clark

    Aurora A depletion reveals centrosome-independent polarization mechanism in C. elegans

    Kerstin Klinkert, Nicolas Levernier, Peter Gross, et al.

    AND

    Centrosome Aurora A gradient ensures a single PAR-2 polarity axis by regulating RhoGEF ECT-2 localization in C. elegans embryos

    Sachin Kotak, Sukriti Kapoor



    Selected by Giuliana Clemente

    Anti-angiogenic effects of VEGF stimulation on endothelium deficient in phosphoinositide recycling

    Amber N Stratman, Olivia M Farrelly, Constantinos M Mikelis, et al.



    Selected by Coert Margadant

    SOL1 and SOL2 Regulate Fate Transition and Cell Divisions in the Arabidopsis Stomatal Lineage

    Abigail R Simmons, Kelli A Davies, Wanpeng Wang, et al.



    Selected by Martin Balcerowicz

    Analysis of the role of Nidogen/entactin in basement membrane assembly and morphogenesis in Drosophila

    Jianli Dai, Beatriz Estrada, Sofie Jacobs, et al.



    Selected by Nargess Khalilgharibi

    Neural crest cells regulate optic cup morphogenesis by promoting extracellular matrix assembly

    Chase Dallas Bryan, Rebecca Lynne Pfeiffer, Bryan William Jones, et al.



    Selected by Ashrifia Adomako-Ankomah

    1

    Simultaneous production of diverse neuronal subtypes during early corticogenesis

    Elia Magrinelli, Robin Jan Wagener, Denis Jabaudon



    Selected by Boyan Bonev

    1

    mRNA localisation in endothelial cells regulates blood vessel sprouting

    Guilherme Costa, Nawseen Tarannum, Shane Herbert



    Selected by Andreas van Impel

    LADL: Light-activated dynamic looping for endogenous gene expression control

    Mayuri Rege, Ji Hun Kim, Jacqueline Valeri, et al.



    Selected by Ivan Candido-Ferreira

    Quantitative, real-time, single cell analysis in tissue reveals expression dynamics of neurogenesis

    Cerys S Manning, Veronica Biga, James Boyd, et al.



    Selected by Teresa Rayon

    The cis-regulatory logic underlying abdominal Hox-mediated repression versus activation of regulatory elements in Drosophila



    Selected by Clarice Hong

    1

    Optogenetic dissection of mitotic spindle positioning in vivo

    Lars-Eric Fielmich, Ruben Schmidt, Daniel J Dickinson, et al.



    Selected by Angika Basant

    1

    Excitable RhoA dynamics drive pulsed contractions in the early C. elegans embryo.

    Jonathan B Michaux, Francois B Robin, William M McFadden, et al.



    Selected by Sundar Naganathan

    A transition from SoxB1 to SoxE transcription factors is essential for progression from pluripotent blastula cells to neural crest cells

    Elsy Buitrago-Delgado, Elizabeth Schock, Kara Nordin, et al.



    Selected by Amanda Haage

    Moving beyond P values: Everyday data analysis with estimation plots

    Joses Ho, Tayfun Tumkaya, Sameer Aryal, et al.



    Selected by Gautam Dey

    1

    Close