Imaging cellular and molecular processes driving cell migration during development

Lead Research Organisation: University of Oxford
Department Name: Physiology Anatomy and Genetics

Abstract

Cell movements play a central role throughout vertebrate embryogenesis. Migration is as fundamental a feature in proper development as proliferation, differentiation, and apoptosis. Yet less is known about cell migration patterns and the molecular mechanisms that govern them then about other aspects of development. By nature, the study of cell migration requires combined knowledge of anatomical landmarks and dynamic migratory patterns. An understanding of such dynamic processes is essential to integrate our increasingly detailed knowledge of genetic regulatory networks into the context of cellular interactions and morphogenesis. Recent technological advances have opened up an exciting new era in developmental biology where we can now address in detail and in real-time how cell behaviour governs the development of the embryo. Our groups are examining such intricately choreographed processes in distinct yet overlapping systems, such as the early mammalian embryo and in the mid-gestation to adult vertebrate nervous system. In order to be able to investigate such cell behaviour in the 3-dimensional context of the organism, we require a microscope with the ability to optically section our samples and obtain high resolution images. Furthermore we require a microscope that can do this over time, as development occurs over time. The state-of-the art equipment requested in this proposal would fulfill these requirements and enable us to achieve our objectives which address fundamental questions about the development of the adult form.

Technical Summary

The Begbie group study sensory ganglia development. The process of forming the cranial sensory ganglia involves the migration of neuroblasts from neurogenic placodes in the surface ectoderm into the embryo. The Begbie group will use genetic constructs in collaboration with the Srinivas lab to characterize the behaviour and mode of neuroblast migration. Further they will determine how this process is coordinated with and by the neural crest cells. The Molnár group studies the migration dynamics of corticofugally projecting subplate neurons using a GFP transgenic mouse line to mark subplate cells and their projections in sub and intracortical circuits during early postnatal development. This group hopes to understand the molecular and cellular mechanisms of neuronal migration and links to human disorders. This work depends on comparing the timing and pattern of migration exhibited by various types of neurons. The Srinivas group studies the molecular and cellular basis for the migration of the AVE, a tissue that specifies and positions the anterior CNS. The group is also conducting a non-invasive lineage analysis of embryonic cells during gastrulation and cardiogenesis using a genetic lineage labeling system and using KikGR mice (a photoconvertable fluorescent protein). The Srinivas group is also generating reporter strains of mice expressing fluorescent Ca2+ reporters to study various roles of this important second messenger in development. The Szele group works on stem cells, neurogenesis, and migration in the postnatal mammalian subventricular zone (SVZ). Using high resolution confocal imaging, the Szele group studies the anatomical localization of subfields and subtypes of cells involved in adult neurogenesis. Using this approach, this group has discovered new patterns of newborn neuron migration. The experiments proposed by this group will determine cellular rules and moleculuar mechanisms that are important for regulating SVZ cell migration.

Publications


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Arnold SJ (2009) Generation and analysis of a mouse line harboring GFP in the Eomes/Tbr2 locus. in Genesis (New York, N.Y. : 2000)
Costantini F (2011) Imaging kidney development. in Cold Spring Harbor protocols
Débarre D (2009) Image-based adaptive optics for two-photon microscopy. in Optics letters
Débarre D (2009) Image-based adaptive optics for two-photon microscopy. in Optics letters
Joyce B (2012) Cell movements in the egg cylinder stage mouse embryo. in Results and problems in cell differentiation
 
Description Our research output from this grant has uncovered a variety of fundamental biological processes, relating to the formation of the earliest specialised cell types in the embryo to how cells migrate in the embryo, helping to shape it.
Exploitation Route Our findings reveal the basic biology underlying embryonic development. It can form the basis of further studies by others that will help for example in using stem cells to generate useful tissues, or to understand how cancer cells migrate and spread, and how to combat this process.
Sectors Education,Healthcare,Pharmaceuticals and Medical Biotechnology
 
Description Our findings have been cited by others, so have been used to inform their research.
First Year Of Impact 2008
Sector Education
Impact Types Societal
 
Description Characterising Epithelial Cell Movements during Anterior Patterning
Amount £509,733 (GBP)
Funding ID BB/J00989X/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 05/2012 
End 10/2015
 
Description Gastrulation in reptiles: Characterisation of turtle and chameleon embryos
Amount £6,000 (GBP)
Funding ID IE121500 
Organisation The Royal Society 
Sector Academic/University
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 06/2013 
End 06/2014
 
Description Gastrulation in reptiles: Characterisation of turtle and chameleon embryos
Amount £6,000 (GBP)
Funding ID IE121500 
Organisation The Royal Society 
Sector Academic/University
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 06/2013 
End 06/2014
 
Description Wellcome Trust Senior Investigator Award
Amount £1,760,000 (GBP)
Funding ID 103788/Z/14/Z 
Organisation The Wellcome Trust Ltd 
Sector Charity/Non Profit
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 01/2015 
End 01/2020
 
Description Wellcome Trust Strategic Award
Amount £2,400,000 (GBP)
Funding ID 105031/C/14/Z 
Organisation The Wellcome Trust Ltd 
Sector Charity/Non Profit
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 10/2014 
End 10/2019
 
Description Wellcome Trust Technology Development Award
Amount £1,500,000 (GBP)
Funding ID 108438/Z/15/Z 
Organisation The Wellcome Trust Ltd 
Sector Charity/Non Profit
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 10/2015 
End 10/2020
 
Description Collaboration with condensed matter physicist Dr John Biggins 
Organisation University of Cambridge
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Pre-implantation embryology and cell fate specification
Collaborator Contribution Quantitative analysis of cellular parameters during early lineage specification
Impact publication: Watanabe et al 2014 multidisciplinary: biology and physics
Start Year 2010
 
Description Collaboration with mathematical biologist Prof Philip Maini 
Organisation University of Oxford
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Collaboration with the director of the Institute for Mathematical Biology at Oxford.
Collaborator Contribution Built computational model of cell migration during early embryonic development
Impact Publication: Trichas et al 2012 Multidisciplinary collaboration: Biology and Mathematics
Start Year 2009
 
Description Tristan Rodriguez 
Organisation Imperial College London (ICL)
Department Imperial College Trust
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Charity/Non Profit 
PI Contribution imaging expertise
Collaborator Contribution molecular embryology expertise
Impact publications: Trichas et al. 2011, Stuckey et al. 2011, Trichas et al 2012
Start Year 2006
 
Description Wolf Reik single cell sequencing 
Organisation Babraham Institute
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Private 
PI Contribution Expertise in early mouse embryogenesis
Collaborator Contribution Expertise single cell sequencing approaches to understand the molecular basis of generation of cellular diversity.
Impact Multi-disciplinary collaboration between developmental biologists, molecular biologists, bioinformatic specialist and mathematical biologists.
Start Year 2014
 
Title 'SilentMark' software for the automated quantification of fluorescent signal in biological material 
Description Karolis Leonavicius (a BBSRC funded DPhil student in my group) created software that uses a novel sampling based approach for the quantification of fluorescence signal in different compartments of the cell. Used on microscopy images of immnuno-fluorescence stained samples, this allows one to quantify the amount of specific proteins in various cellular compartments. Importantly, the software also allows one to relate the fluorescence intensity to the spatial landmarks with the sample, which is particularly important in developmental biology applications. 
Type Of Technology Software 
Year Produced 2016 
Impact Thie software has been used by us in our research and by a collaborator in an unrelated project. We are in the process of publishing this software so that the wider community has access to it. 
 
Description BBC Radio interview on first heart beat 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Following a recent publication (Tyser et al. eLIFE 2016), I was invited to speak briefly on the local BBC Oxford radio station. The broadcast aired during in the early evening with the target audience being commuters returning home (the 'Drivetime Show'). The brief 5 minute interview covered our findings about the earliest contractile activity in the embryonic heart.
Year(s) Of Engagement Activity 2016
 
Description Cheny School Science Festival 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Participation in a Science Festival organised by Cheny Secondary School. In collaboration with Prof Mathilda Mommersteeg form my department and Micron Oxford (a Wellcome Trust funded advanced imaging unit based in the adjoining Dept of Biochemistry) we had a table at the festival where we described out work on imaging embryonic morphogenesis over a period of approximately 5 hours. The co-ordination and planning of our activities was done by a postdoctoral fellow in my group Dr. Tomoko Watanabe, who was funded previously through a BBSRC grant and is currently funded through a Wellcome Trust grant.

Acting on experience gained from our previous activity (Oxford Science Festival 2016), we were able to refine our activities for this event, as well as trial two additional activities. A student in my group created an 'World' in the computer game 'Minecraft' depicting different stages of heart development, as well as a maze through a developing heart. This attracted much interest from the secondary school students who visited our table. We also had on display cultured mouse caridomyocytes that visitors could watch beating using a microscope. In addition to these, as with our last event we had: 3D print-outs of different stages of embryonic heart development; fixed specimens of mouse, chick and zebrafish embryos; a microscope with live zebrafish embryos; a 3D printer printing models of embryos; a game we designed where children use stickers to fill in missing stages in development.
Year(s) Of Engagement Activity 2017
 
Description OSF2016 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Participation in the Oxford Science Festival. In collaboration with two colleagues from my department (Profs. Jo Begbie and Mathilda Mommersteeg) and Micron Oxford (a Wellcome Trust funded advanced imaging unit based in the adjoining Dept of Biochemistry) my group had a table over two full days (Saturday and Sunday) in which we described out work on studying embryonic morphogenesis. The co-ordination and planning of our activities was done by a postdoctoral fellow in my group Dr. Tomoko Watanabe, who was funded previously through a BBSRC grant and is currently funded through a Wellcome Trust grant.

Items used to facilitate engagement included: 3D print-outs of different stages of embryonic heart development; fixed specimens of mouse, chick and zebrafish embryos; a microscope with live zebrafish embryos; a 3D printer printing models of embryos; a game we designed where children use stickers to fill in missing stages in development; and moulding clay that visitors could use to make models of embryos.

Participation in this resulted in an invitation to another science festival at a local secondary school.
Year(s) Of Engagement Activity 2016