Imaging cellular and molecular processes driving cell migration during development
Lead Research Organisation:
University of Oxford
Department Name: Human 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
Berenjeno I
(2017)
Oncogenic PIK3CA induces centrosome amplification and tolerance to genome doubling
in Nature Communications
Bilanges B
(2017)
Vps34 PI 3-kinase inactivation enhances insulin sensitivity through reprogramming of mitochondrial metabolism.
in Nature communications
Chen CM
(2014)
Detecting cardiac contractile activity in the early mouse embryo using multiple modalities.
in Frontiers in physiology
Costantini F
(2011)
Imaging kidney development.
in Cold Spring Harbor protocols
Costantini F
(2011)
Dissection of embryonic mouse kidney, culture in vitro, and imaging of the developing organ.
in Cold Spring Harbor protocols
Débarre D
(2009)
Image-based adaptive optics for two-photon microscopy.
in Optics letters
Jesacher A
(2009)
Adaptive harmonic generation microscopy of mammalian embryos
in Optics Letters
Joyce B
(2012)
Cell movements in the egg cylinder stage mouse embryo.
in Results and problems in cell differentiation
Leonavicius K
(2020)
Spatial protein analysis in developing tissues: a sampling-based image processing approach.
in Philosophical transactions of the Royal Society of London. Series B, Biological sciences
Marshall G
(2010)
Three-dimensional imaging of direct-written photonic structures
Marshall GD
(2011)
Three-dimensional imaging of direct-written photonic structures.
in Optics letters
Raimondo JV
(2013)
A genetically-encoded chloride and pH sensor for dissociating ion dynamics in the nervous system.
in Frontiers in cellular neuroscience
Royer C
(2014)
ASPP2 links the apical lateral polarity complex to the regulation of YAP activity in epithelial cells.
in PloS one
Srinivas S
(2017)
A Tale of Division and Polarization in the Mammalian Embryo.
in Developmental cell
Stower M
(2015)
Bi-modal strategy of gastrulation in reptiles
in Developmental Dynamics
Stower MJ
(2018)
The Head's Tale: Anterior-Posterior Axis Formation in the Mouse Embryo.
in Current topics in developmental biology
Thayil A
(2011)
Self calibration of sensorless adaptive optical microscopes
in Journal of the European Optical Society: Rapid Publications
Thayil A
(2011)
Long-term imaging of mouse embryos using adaptive harmonic generation microscopy.
in Journal of biomedical optics
Trichas G
(2008)
Use of the viral 2A peptide for bicistronic expression in transgenic mice.
in BMC biology
Trichas G
(2012)
Multi-cellular rosettes in the mouse visceral endoderm facilitate the ordered migration of anterior visceral endoderm cells.
in PLoS biology
Trichas G
(2011)
Nodal dependent differential localisation of dishevelled-2 demarcates regions of differing cell behaviour in the visceral endoderm.
in PLoS biology
Tyser RC
(2016)
Calcium handling precedes cardiac differentiation to initiate the first heartbeat.
in eLife
Watanabe T
(2010)
Characterisation of the dynamic behaviour of lipid droplets in the early mouse embryo using adaptive harmonic generation microscopy.
in BMC cell biology
Watanabe T
(2014)
Limited predictive value of blastomere angle of division in trophectoderm and inner cell mass specification.
in Development (Cambridge, England)
| 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 |
| Start | 04/2012 |
| 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 | Charity/Non Profit |
| Country | United Kingdom |
| Start | 05/2013 |
| 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 | Charity/Non Profit |
| Country | United Kingdom |
| Start | 05/2013 |
| End | 06/2014 |
| Description | Wellcome Trust Senior Investigator Award |
| Amount | £1,760,000 (GBP) |
| Funding ID | 103788/Z/14/Z |
| Organisation | Wellcome Trust |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 01/2015 |
| End | 01/2020 |
| Description | Wellcome Trust Strategic Award |
| Amount | £2,400,000 (GBP) |
| Funding ID | 105031/C/14/Z |
| Organisation | Wellcome Trust |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 09/2014 |
| End | 10/2019 |
| Description | Wellcome Trust Technology Development Award |
| Amount | £1,500,000 (GBP) |
| Funding ID | 108438/Z/15/Z |
| Organisation | Wellcome Trust |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 09/2015 |
| End | 10/2020 |
| Description | Collaboration with condensed matter physicist Dr John Biggins |
| Organisation | University of Cambridge |
| Country | United Kingdom |
| 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 |
| 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 |
| Department | Imperial College Trust |
| Country | United Kingdom |
| 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 |
| Sector | Academic/University |
| 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 | Oxford Science Festival 2016 |
| 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 |
| Description | Oxford Science Festival 2017 |
| 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 | For the second consecutive year, 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. As before, 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. This year, we had a new display item, a 'World of Minecraft' rendering of the inside of a mouse heart. This attracted much interest from a segment of the public in the age range of 8-18 year old that we have had only limited success engaging with in the past. |
| Year(s) Of Engagement Activity | 2017 |