2019 Fiscal Year Research-status Report
Molecular regulation of blood vessel size by endothelial cells-the role of Marcksl1.
| Project/Area Number |
19K06651
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Phng LiKun 国立研究開発法人理化学研究所, 生命機能科学研究センター, チームリーダー (70794098)
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| Project Period (FY) |
2019-04-01 – 2022-03-31
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| Keywords | Endothelial cell / Angiogenesis / Actin / Marcksl1 |
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| Outline of Annual Research Achievements |
The tubulogenesis of blood vessels is a multistep process that requires the establishment of apical-basal polarity, the remodeling of cell-cell adhesion and endothelial cell (EC) shape changes. EC shape changes is driven by intrinsic force generation that is actomyosin-dependent, and by extrinsic forces generated by haemodynamics. Live imaging revealed that haemodynamic forces deform the apical membrane to generate inverse blebs that, when uncontrolled, are detrimental to lumen formation. Here, we identified a role of Marcksl1, an actin bundling protein, in modulating EC response to haemodynamic forces. The increase in Marcksl1 expression level resulted in ectopic membrane blebbing, changes in EC shape and increased vessel diameter while reduced expression level decreased vessel diameter.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
We have successfully generated zebrafish genetic mutants to investigate the loss of Marcksl1a and Marcksl1b function during development. We have been able to examine EC shape and vessel network structure by combining the use of transgenic zebrafish in which ECs are fluorescently labelled with confocal live imaging. Furthemore, we have established an in vitro EC system using human umbilical vein endothelial cells (HUVECs) to examine the actin cytoskeleton at high resolution after perturbing the levels of Marcksl1.
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| Strategy for Future Research Activity |
The future plan is to determine the mechanism by which Marcksl1 regulates EC shape. Here, we plan to quantitatively examine the organisation of actin cytoskeleton in wildtype, Marcksl1-deficient and Marcksl1-overexpressing ECs. Actin cytoskeleton will be visualized by using either Lifeact-EGFP or other actin biosensors and super resolution imaging. As we anticipate that in vivo super resolution imaging to be challenging, we will also analyse the organisation of actin in ECs in culture.
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| Causes of Carryover |
Funding from the grant was used to attend two international conferences (USA and Australia) in FY2019.
In FY2020, I plan to use the remainder of the FY2019 funding and FY2020 funding on attending meetings, purchasing research consumables/equipment and publication costs (miscallaneous).
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