2021 Fiscal Year Research-status Report
A mechanical study of lumen formation through membrane bleb regulation under hemodynamic influence
| Project/Area Number |
20K20190
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
MaungYe SweSoe 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, 基礎科学特別研究員 (60866408)
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| Project Period (FY) |
2020-04-01 – 2023-03-31
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| Keywords | Microhemodynamics / Hematocrit asymmetry / Wall shear stress / Vascular morphogenesis / Membrane blebbing / RBC mechanics / Endothelial mechanics / Vascular mechanics |
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| Outline of Annual Research Achievements |
1) A semi-automated imaging and calculation method was established for the high throughput analysis of growth and hemodynamic trends in zebrafish populations. This work provides the background information to lumen formation and remodeling in terms of the overarching spatiotemporal systemic hemodynamic trends and vascular network level adaptations in response to local lumen dynamics. A paper titled "High-throughput imaging of blood flow reveals developmental changes in distribution patterns of hemodynamic quantities in developing zebrafish" has been submitted to Frontiers in Physiology on the 23rd Feb 2022. The manuscript has passed the review process and is awaiting official publication.
2) To measure the flow forces during apical membrane blebbing in a developing lumen, I have developed a computational fluid dynamics(CFD) model of blood flow in the zebrafish. Using this model, I discuss how local morphological changes alter network flow, wall shear stress and pressure distributions between wild type and mutant phenotypes regulating vessel caliber such as Marcksl1 overexpression (OE) (where extensive apical blebbing occurs) versus Marcksl1 knock-out (KO) phenotype (where vessel diameters are reduced in concomitance with the bleb inhibition). The results are being prepared for submission to a peer-reviewed journal.
3) Experiments with caveolin marked zebrafish were performed to assess membrane tension during apical blebbing. Preliminary results indicate promising potential of caveolin marker to study membrane tension in apical membrane during bleb-induced lumen formation.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
A replacement methodology for membrane tension measurement was required due to the lack of success with FlipTR tension probe originally planned for measuring the tension. Additional experiments to verify caveolin markers for this purpose delayed the research pipeline slightly as a result.
Publication activity involving the hemodynamics measurement strategies also took more time than initially planned.
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| Strategy for Future Research Activity |
1) Submit paper on CFD analysis of network hemodynamics.
2) Complete experiments of apical membrane tension measurement and actomyosin dynamics quantification using caveolin.
3) Construct 2-layer membrane computational model of endothelial cell undergoing apical membrane blebbing in the vascular environment
4) Construct the computational models of actomyosin transport and membrane bleb regulation
5) Combine models to demonstrate lumen expansion under the multiple factors of actomyosin assembly and hemodynamic force variation due to
network flow effects
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| Causes of Carryover |
Key experiments were delayed due to a replacement experiment methodology being verified. This meant costs of the main experiments were largely not incurred.
Additionally, key data for the next stage of model building was delayed and the purchase of a new workstation computer to undertake the model simulation was postponed.
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