Thesis
Computational methods for light sheet fluorescence microscopy: towards a quantitative analysis of AVE migration in the mouse
- Abstract:
-
Collective migration in epithelial tissues is required for many developmental processes, wound healing, and cancer, and is conserved across vertebrates. It is a process that has been difficult to study for 3D epithelia in vivo as the large timelapse datasets required present significant computational challenges for quantitative analysis. An important example of collective migration in an epithelial context is the migration of Anterior Visceral Endoderm (AVE) cells in the mouse embryo. From E5.5, this subgroup of cells in the visceral endoderm monolayer migrate from the distal tip, specifying the anterior-posterior axis by patterning the underlying epiblast which will later give rise to the embryonic lineage. AVE induction and migration have yet to be fully characterised and understood, and provide an attractive in vivo system to study the biological function of collective migration due to the relatively small cell numbers at this stage and the experimental accessibility of the visceral endoderm tissue. In this thesis, a new computational framework is developed to analyse and extract quantitative information from light sheet fluorescence microscopy (LSFM) 3D timelapse datasets of E5.5 mouse embryos during AVE migration. The proposed framework initially isolates the migratory cell movement, and secondly extracts the apical surface data to simplify the 3D volume to a 2D characterisation problem. Applying this computational framework on LSFM datasets from 3 fluorescent reporter mouse lines, I demonstrate the first comprehensive tracking of individual cells and cell divisions in the entire visceral endoderm, the first study of cell division angles, and the movement of cells in the underlying epiblast. Altogether these results reveal a model of AVE migration as an active process that drives large-scale tissue deformation and changes in morphology independent of cell proliferation in the VE, and presents evidence that suggests the A-P axis could be pre-defined.
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Authors
Contributors
- Institution:
- University of Oxford
- Division:
- MSD
- Department:
- Physiology Anatomy & Genetics
- Research group:
- Srinivas
- Role:
- Contributor, Supervisor
- ORCID:
- 0000-0002-4892-7687
- Institution:
- University of Oxford
- Division:
- MSD
- Department:
- NDM
- Sub department:
- Oxford Ludwig Institute
- Research group:
- Rittscher Group
- Role:
- Contributor
- ORCID:
- 0000-0003-4463-1165
- Institution:
- University of Liverpool
- Role:
- Contributor
- ORCID:
- 0000-0001-7395-2879
- Institution:
- University of Oxford
- Division:
- MSD
- Department:
- NDM
- Research group:
- Rittscher Group
- Oxford college:
- Harris Manchester College
- Role:
- Supervisor
- ORCID:
- 0000-0002-8528-8298
- Institution:
- University of Oxford
- Division:
- MSD
- Department:
- Physiology Anatomy & Genetics
- Research group:
- Srinivas
- Oxford college:
- Jesus College
- Role:
- Supervisor
- ORCID:
- 0000-0001-5726-7791
- Funding agency for:
- Hathrell, H
- Grant:
- 109100/Z/15/Z
- Programme:
- Chromosome and Developmental Biology Doctoral Training Centre
- Type of award:
- DPhil
- Level of award:
- Doctoral
- Awarding institution:
- University of Oxford
- Language:
-
English
- Keywords:
- Subjects:
- Deposit date:
-
2020-09-23
Terms of use
- Copyright holder:
- Hathrell, H
- Copyright date:
- 2020
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