Somitic muscle growth: a new model for the role of force in morphogenesis

Lead Research Organisation: King's College London
Department Name: Randall Div of Cell and Molecular Biophy

Abstract

Skeletal muscle makes up 40% of our body mass and its maintenance is essential to a good quality of life. In muscle diseases, cancer, chronic kidney failure, AIDS and rheumatoid arthritis, and particularly in older or hospitalised people, muscle wasting is a serious problem. The physical force of exercise prevents wasting by promoting muscle growth. How the body decides what is an appropriate amount of muscle is unknown, but a likely mechanism is feedback from the force of muscle contraction. We want to find out how muscle tissue detects force and responds by growing. In the past, we have discovered how early muscle is made in similar ways in fish, mice and people. Now, we want to study the role of force in later muscle growth. So our first aim is determine how muscle normally grows and how force and other signals influence growth. We will use young zebrafish because they are a) transparent, so that we can study muscle growth in the living animal, b) easy to manipulate genetically, so we can find out what genes control growth, c) small and accessible, so we can measure and impose forces in the living muscle. These things are more difficult to do in mammals. Our second aim is to understand how muscle detects force and uses the information to orchestrate growth. Muscle fibre ends transmit force to adjacent tissues by means of a complex set of proteins that form attachments to neighbouring cells. Evidence leads us to hypothesise that these attachments are also force-detectors that initiate the process of growth in response to certain kinds of contractile activity. We will study the composition and dynamics of the protein complex at fibre ends and test its role in force detection and growth response. Muscle is continually damaged during life and is normally repaired by stem cells, which also contribute to growth. Our third aim is to understand how muscle stem cells form, where they reside, and how force or other signals regulate their behaviour to coordinate muscle growth and repair. Bone, heart and skin also respond to force by altered growth. Understanding the force detection and response systems in skeletal muscle is likely to illuminate the role of force in biology and regenerative medicine more generally.

Technical Summary

The role of mechanical force in development is poorly understood. Forces drive morphogenesis, the emergence of biological form. Force also provides information to cells about their situation within a tissue. The aim of this proposal is to develop zebrafish skeletal muscle as a model for the study of force in biological systems, focusing on the role of force in morphogenesis. Specifically, we want to understand how muscle senses force and responds by changing the number and size of muscle fibres. Work from a small group of labs, including ours, has developed the zebrafish somite as an advanced model of early myogenesis, revealing mechanisms controlling diversification of muscle and associated stem cells. Our recent findings indicate that force regulates a later phase of somitic muscle growth and implicate muscle fibre end attachments as sites at which force is sensed. The evidence suggests that contractile activity and the force it generates control increase in both fibre number and volume. By describing the normal process of muscle growth at single cell resolution in live vertebrates, something not previously achieved, we are in a position to investigate exactly how the signals and forces that regulate muscle growth work. We will ask where force is sensed, what biochemical pathways are required to transduce force signals and which cells are the ultimate targets of such signals. Fibre end attachments are regions where muscle exerts force and simultaneously grows. Evidence leads us to hypothesise that fibre end attachments act as force sensors, feeding back information to the cell nucleus and to adjacent cells to regulate muscle growth. Understanding the role of force in muscle growth is an essential first step in developing optimised methods to prevent weakening in ageing-related muscle wasting, heart disease, cachexia and other diseases affecting muscle. It will also impact on sports science, meat production and regenerative medicine generally. The specific aims of this proposal are to study how muscle form arises from interactions of cells and forces by analysing:
1. The locations and mechanisms of somite growth in response to force.
2. Muscle fibre attachments and their role in force-sensing.
3. The origin, niche and regulation by force of muscle stem cells.

Publications

10 25 50
 
Description AMM Funding
Amount £180,000 (GBP)
Organisation Monaco Association against Duchenne Muscular Dystrophy 
Sector Charity/Non Profit
Country Monaco
Start 01/2009 
End 12/2011
 
Description BBSRC Project Grant
Amount £363,618 (GBP)
Funding ID BB/K010115. 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2013 
End 10/2016
 
Description BHF Centre Pump Priming Grant
Amount £60,248 (GBP)
Organisation King's College London 
Department BHF Centre of Research Excellence
Sector Academic/University
Country United Kingdom
Start 07/2012 
End 06/2013
 
Description BHF PhD Studentship
Amount £115,447 (GBP)
Organisation King's College London 
Department BHF Centre of Research Excellence
Sector Academic/University
Country United Kingdom
Start 10/2012 
End 09/2015
 
Description BHF Project Grant
Amount £265,335 (GBP)
Funding ID PG/14/12/30664 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 07/2014 
End 06/2017
 
Description MRC Programme Grant 2016-21
Amount £1,800,000 (GBP)
Funding ID MR/N021231/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 06/2016 
End 05/2021
 
Description MRC Research Project
Amount £1,649,876 (GBP)
Funding ID MR/K015664/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 01/2013 
End 12/2017
 
Description Marie Curie IEF Fellowship
Amount £260,000 (GBP)
Organisation Marie Sklodowska-Curie Actions 
Sector Academic/University
Country Global
Start 01/2010 
End 12/2012
 
Description Wellcome Trust Equipment
Amount £707,233 (GBP)
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start  
End 12/2012
 
Title DNA clones 
Description Diverse clone constrcuts 
Type Of Material Technology assay or reagent 
Year Produced 2006 
Provided To Others? Yes  
Impact Many publications 
 
Title FRAP analysis in vivo 
Description New methods of imaging and software for processing FRAP analyses performed in live tissue were developed. Software was made publicly available free of charge associated with Open Access publication in eLife. DOI: 10.5061/dryad.qg8dt 
Type Of Material Technology assay or reagent 
Year Produced 2015 
Provided To Others? Yes  
Impact unknown as yet 
 
Title Genetically modified zebrafish 
Description Numerous transgenic or mutant zebrafish strains 
Type Of Material Model of mechanisms or symptoms - non-mammalian in vivo 
Provided To Others? No  
Impact Novel biological insight in muscle and heart biology 
 
Title MRF mutants 
Description four lines of mutant zebrafish 
Type Of Material Model of mechanisms or symptoms - non-mammalian in vivo 
Year Produced 2009 
Provided To Others? Yes  
Impact improved understanding of myogenesis 
 
Title Monoclonal Antibodies 
Description Antibodies to myosin 
Type Of Material Antibody 
Year Produced 2006 
Provided To Others? Yes  
Impact many publications 
 
Description BHF Pump Priming Initiative 
Organisation King's College London
Department BHF Centre of Research Excellence
Country United Kingdom 
Sector Academic/University 
PI Contribution Dr Yaniv Hinits modified zebrafish BACs and made transgenic fish for distribution to partners
Collaborator Contribution Financial contribution to Y Hinits salary
Impact Markers for specific populations of cardiovacscular system cells in zebrafish
Start Year 2012
 
Description BHF Pump Priming Initiative 
Organisation University of Edinburgh
Department Centre for Cardiovascular Science
Country United Kingdom 
Sector Academic/University 
PI Contribution Dr Yaniv Hinits modified zebrafish BACs and made transgenic fish for distribution to partners
Collaborator Contribution Financial contribution to Y Hinits salary
Impact Markers for specific populations of cardiovacscular system cells in zebrafish
Start Year 2012
 
Description BHF Pump Priming Initiative 
Organisation University of Oxford
Department BHF Centre of Research Excellence
Country United Kingdom 
Sector Academic/University 
PI Contribution Dr Yaniv Hinits modified zebrafish BACs and made transgenic fish for distribution to partners
Collaborator Contribution Financial contribution to Y Hinits salary
Impact Markers for specific populations of cardiovacscular system cells in zebrafish
Start Year 2012
 
Description Borycki Shh collab 
Organisation University of Sheffield
Department Department of Biomedical Science
Country United Kingdom 
Sector Academic/University 
PI Contribution We performed experiments and provided knockout mice
Collaborator Contribution They provided other data
Impact Anderson et al 2012, on which V. Williams form my lab is joint first author.
Start Year 2010
 
Description Cannabinoid Receptor signalling 
Organisation King's College London
Department Randall Division of Cell & Molecular Biophysics
Country United Kingdom 
Sector Academic/University 
PI Contribution Provided advice, reagents, fish and experiments.
Collaborator Contribution Student worked to mutate two gene in zebrafish using CRISPR/Cas9
Impact Manuscript published
Start Year 2014
 
Description Dermomyotome 
Organisation Wesleyan University
Country United States 
Sector Academic/University 
PI Contribution Ideas, Writing and experiments
Collaborator Contribution ideas and writing and data
Impact 16409387
 
Description Dystrophin papers 
Organisation King's College London
Department School of Medicine KCL
Country United Kingdom 
Sector Academic/University 
PI Contribution experimental work and writing
Collaborator Contribution experiments
Impact 18042440 17233888
Start Year 2006
 
Description Function of RNA binding proteins 
Organisation King's College London
Department Division Analytical and Environmental Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution We deleted two genes in the zebrafish
Collaborator Contribution Advice, stundet recruitment
Impact Manuscript in preparation.
Start Year 2014
 
Description Garcia 
Organisation Pierre and Marie Curie University - Paris 6
Country France 
Sector Academic/University 
PI Contribution Experimental research
Collaborator Contribution Provision of reagents
Impact Publciation submiited
Start Year 2011
 
Description Halevy 
Organisation Hebrew University of Jerusalem
Department Department of Animal Sciences
Country Israel 
Sector Academic/University 
PI Contribution GM mice and resource during research visit
Collaborator Contribution publication
Impact publicaton
Start Year 2009
 
Description Head muscles Patel 
Organisation University of Reading
Department School of Biological Sciences Reading
Country United Kingdom 
Sector Academic/University 
PI Contribution Hinits and Minchin designed, peformed and provided experimental data and helped write the manuscript
Collaborator Contribution Publication
Impact see publication
Start Year 2009
 
Description ICE 
Organisation Pierre and Marie Curie University - Paris 6
Department UMR 787 (Institute of Myology)
Country France 
Sector Academic/University 
PI Contribution We are anaylsing dynamics of short Dystrophins in zebrafish to understand their utility in gene therapy for Duchenne Muscular Dystrophy
Collaborator Contribution they cloned and mutated human Dystrophin
Impact ongoing
Start Year 2009
 
Description MRF mutant fish 
Organisation Howard Hughes Medical Institute
Country United States 
Sector Charity/Non Profit 
PI Contribution The entire work analysis of the mutant fish and writing the publication
Collaborator Contribution mutant fishdata
Impact see publication
Start Year 2009
 
Description MRF mutant fish 
Organisation University of Nottingham
Department School of Biosciences
Country United Kingdom 
Sector Academic/University 
PI Contribution The entire work analysis of the mutant fish and writing the publication
Collaborator Contribution mutant fishdata
Impact see publication
Start Year 2009
 
Description Molinari 
Organisation University of Modena and Reggio Emilia
Department Department of Biomedical Sciences
Country Italy 
Sector Academic/University 
PI Contribution Hosted PhD student and we performed experiments in live zebrafish
Collaborator Contribution Performed biochemical analyses on Mef2
Impact ongoing
Start Year 2009
 
Description PNAS paper 
Organisation University College London
Department Biosciences
Country United Kingdom 
Sector Academic/University 
PI Contribution Set of experiments and longstanding intellectual input
Collaborator Contribution Majority of research work in paper
Impact 19020093, press coverage
 
Description Sear 
Organisation University of Surrey
Country United Kingdom 
Sector Academic/University 
PI Contribution Experimental data and advice
Collaborator Contribution Collaboration to mathematically model and write software
Impact paper submitted
Start Year 2012
 
Title Commericalisation of Monoclonal antibodies 
Description monoclonal antibody reagents 
IP Reference  
Protection Protection not required
Year Protection Granted 2012
Licensed Yes
Impact MRC Technology decided not to continue license agreement with Stanford University
 
Title FRAP software 
Description Software for analysis of FRAP data 
Type Of Technology Software 
Year Produced 2014 
Open Source License? Yes  
Impact unknown as yet 
 
Description Coverage on KCL website 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Type Of Presentation Poster Presentation
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Website coverage of our Paper in PLOS Biology

unknown except feedback form colleagues
Year(s) Of Engagement Activity 2013
URL http://www.kcl.ac.uk
 
Description NIG Collaborative Visit 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact 50 NIG scientists attended talk
Collaborative research initiated

Discovered many fish lines useful for UK research
Year(s) Of Engagement Activity 2013
URL http://www.nig.ac.jp/english/index.html
 
Description School Visit London 2012 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Workshop Facilitator
Geographic Reach Local
Primary Audience Schools
Results and Impact 150 pupils year 10 heard presentation and discussed Ethics of Stem Cells

very active engagement of students
Year(s) Of Engagement Activity 2012
 
Description School visit 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Group of Y12 students came to learn about research
Year(s) Of Engagement Activity 2015,2016,2017,2018
 
Description School visit 2013 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Schools
Results and Impact 120 pupils discussed the ethics of the use of animals and stem cell medicine in their GCSE RE classes

Extensive Q+A session after my ppt presentations
Year(s) Of Engagement Activity 2013
 
Description School visit 2014 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact lab visit and discussion

na
Year(s) Of Engagement Activity 2014
 
Description Student internship 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Work experience Y9 and Y12 students
Year(s) Of Engagement Activity 2015,2016,2017
 
Description Zebrafish Facility Opening 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Type Of Presentation Workshop Facilitator
Geographic Reach National
Primary Audience Policymakers/politicians
Results and Impact Lord Taylor of Holbeach, Parliamentary Under Secretary of State for Criminal Information, opened the new KCL Zebrafish facility accompanied by head of Animal Research section of Home Office

Press release and further fact finding visits from HO staff
Year(s) Of Engagement Activity 2013