Investigation of the role of the primary cilia and centrosome during signalling events in development using the chicken mutant talpid3
Lead Research Organisation:
University of Edinburgh
Department Name: The Roslin Institute
Abstract
It is thought that every cell in our bodies has a small antenna poking out of it, called a primary cilia. For many years biologist thought that the primary cilia was no more than a vestigial organ, something left behind from when we were single-celled organisms swimming through the oceans using a flagella (the type of tail sperm and other motile cells use to move). However now it seems that the primary cilia is more than just a stumpy tail, it is in fact essential to normal development of the embryo. Clues as to the importance of cilia, first came from investigation into human diseases in which motile cilia were absent or impaired. Not surprisingly, these people often suffer from respiratory and reproductive problems, as they don't have cilia to move mucus, eggs or sperm. In addition it was found that many of these people also have situs inversus, a condition when the internal organs such as the heart are found on the wrong side of the body. This suggested that cilia somehow play an important role in the development of the embryo, somehow controlling which side organs should form on. To understanding how cilia could control such important events, recent research has aimed to determine what cilia consist of. Using many different techniques including studying mouse and human mutations which result tell-tale cilia syndromes such as the development of extra fingers and kidney problems, scientists have begun to understand which genes are needed to form normal cilia. By studying precisely what goes wrong in animals which lack cilia, we now understand that cilia are required to control important intracellular signalling events, particularly in pathways important in development and cancers known as the Hedgehog and Wnt pathways. Using high powered imaging techniques, researchers have been able to look inside cilia and have found that some important proteins become highly concentrated in cilia, using cilia as a special compartment in which vital protein interactions can take place. But we have little idea how this occurs. We study a chicken flock in which some of the embryos fail to hatch. It was discovered that these embryos die in the egg from serious developmental disorders. By examining the anatomy of these embryos known as talpid3, we discovered that important signalling events in the Hedgehog signalling pathway, go awry, leaving embryos with central nervous system, limb, head, blood vessel and bone defects. Excitingly when the defects in talpid3 embryos are compared to those in humans and mice in which cilia are defective there are many areas of similarity and it now seems talpid3 embryos also have defective cilia. Furthermore the gene which causes talpid3, localises in the centrosome, a structure essential for normal cilia development. Talpid3 embryos offer a unique approach to studying cilia function which is difficult to undertake in mice. Because embryos develop outside the mother (in the egg) it is possible to manipulate embryos while they are still developing. We plan to use talpid3 embryos to examine which genes are important in cilia growth, by over expressing or reducing expression of genes known to be important in cilia formation or Hedgehog signalling in the embryos and then examining cilia formation, or patterning of the nervous system controlled by Hedgehog signalling. We can also use both these embryo manipulations and cell culture techniques to see where proteins localise in the cell. Talpid3 embryos also have one other interesting defect, in that they don't form normal veins. Although this could be because of the abnormal Hedgehog signalling in talpid3 we would also like to investigate if this is due to another function of cilia, which can sense mechanical stresses such as flow. We think it is possible that cilia not only act as a focal point for signalling in the cells but may act to coordinate the many varied signalling events that determine how we develop.
Technical Summary
Primary non-motile cilia (9+0) and their associated centrosomal components, the centriole are essential components of the Wnt, PDGF and Hedgehog (Hh) signalling pathways. Cilia are also important mechanosensory organelles. Defects in genes associated with cilia formation result in a wide range of human developmental disorders, characterised by phenotypes such as central nervous system disorders, kidney failure, blindness and polydactyly. It is known that components of signalling pathways are preferentially concentrated in the cilia but it is unclear how this protein sorting is achieved. Talpid3 is a chicken mutant with a defect in KIAA0586, a gene associated with the centrosome. Talpid3 embryos lack cilia, have developmental defects which resemble phenotypes seen in the human ciliopathies and have a well characterized Hh signalling defect. As developing chicken embryos are readily available and easily to experimentally manipulate, we will use this ciliary mutant in order to understand the mechanisms involved in cilia based intracellular signalling. Using in vitro and in ovo complementation experiments and co-localisation studies, we will observe the effect on cilia and Hh signalling, by manipulating both known components of the Hh pathway and genes involved in ciliogenesis. We will also use this approach to test candidate genes identified as potentially involved in Hh signalling/ciliogenesis in a previous analysis. The mechanosensory role of cilia in the kidney is clear, but the contribution of mechanical stimulation as a driver of morphogenesis is less well understood, although it is known that blood flow contributes to the patterning of vasculature. Talpid3 embryos have defective vascular patterning. We will investigate how the loss of cilia contributes to this phenotype. This work may elucidate the way in which cilia co-ordinate these diverse signalling events to direct morphogenesis.
People |
ORCID iD |
Megan Davey (Principal Investigator) |
Publications
Bangs F
(2011)
Generation of mice with functional inactivation of talpid3, a gene first identified in chicken.
in Development (Cambridge, England)
Bangs F
(2009)
16-P002 The developmental mutant talpid3 lacks primary cilia
in Mechanisms of Development
Bangs F
(2010)
Identification of genes downstream of the Shh signalling in the developing chick wing and syn-expressed with Hoxd13 using microarray and 3D computational analysis.
in Mechanisms of development
Burns A
(2009)
13-P093 Defects in gut and enteric nervous system development in talpid3 mutant chicken
in Mechanisms of Development
Cruz C
(2010)
Foxj1 regulates floor plate cilia architecture and modifies the response of cells to sonic hedgehog signalling.
in Development (Cambridge, England)
Davey MG
(2014)
Loss of cilia causes embryonic lung hypoplasia, liver fibrosis, and cholestasis in the talpid3 ciliopathy mutant.
in Organogenesis
Delalande JM
(2021)
TALPID3/KIAA0586 Regulates Multiple Aspects of Neuromuscular Patterning During Gastrointestinal Development in Animal Models and Human.
in Frontiers in molecular neuroscience
Dunn IC
(2011)
The chicken polydactyly (Po) locus causes allelic imbalance and ectopic expression of Shh during limb development.
in Developmental dynamics : an official publication of the American Association of Anatomists
Fraser AM
(2019)
TALPID3 in Joubert syndrome and related ciliopathy disorders.
in Current opinion in genetics & development
Garceau V
(2010)
Pivotal Advance: Avian colony-stimulating factor 1 (CSF-1), interleukin-34 (IL-34), and CSF-1 receptor genes and gene products.
in Journal of leukocyte biology
Johnson EJ
(2014)
Direct functional consequences of ZRS enhancer mutation combine with secondary long range SHH signalling effects to cause preaxial polydactyly.
in Developmental biology
May-Simera HL
(2016)
Loss of MACF1 Abolishes Ciliogenesis and Disrupts Apicobasal Polarity Establishment in the Retina.
in Cell reports
Patthey C
(2017)
Evolution of the functionally conserved DCC gene in birds.
in Scientific reports
Pinkham J
(2009)
11-P012 Left/right axis specification in the cilia mutant talpid3
in Mechanisms of Development
Rainger J
(2017)
A recurrent de novo mutation in ACTG1 causes isolated ocular coloboma.
in Human mutation
Schock EN
(2016)
Utilizing the chicken as an animal model for human craniofacial ciliopathies.
in Developmental biology
Stephen L
(2013)
Failure of centrosome migration causes a loss of motile cilia in talpid 3 mutants
in Developmental Dynamics
Stephen LA
(2013)
Failure of centrosome migration causes a loss of motile cilia in talpid(3) mutants.
in Developmental dynamics : an official publication of the American Association of Anatomists
Stephen LA
(2014)
The chicken left right organizer has nonmotile cilia which are lost in a stage-dependent manner in the talpid(3) ciliopathy.
in Genesis (New York, N.Y. : 2000)
Wong F
(2013)
eChickAtlas: an introduction to the database.
in Genesis (New York, N.Y. : 2000)
Yin Y
(2009)
The Talpid3 gene (KIAA0586) encodes a centrosomal protein that is essential for primary cilia formation.
in Development (Cambridge, England)
Title | Chicken Wearing SOX |
Description | Image submitted to RPS International Images for Science 2015 |
Type Of Art | Image |
Year Produced | 2016 |
Impact | Public engagement |
URL | http://www.rps.org |
Description | TALPID3 protein is a centrosomal protein It is postulated through analysis of TALPID3 homologues that TALPID3 has a Eumetazoan origin TALPID3 has 2 essential coiled coiled domains TALPID3 has 1 essential region of intrisic disorder TALPID3 is essential for motile and non-motile cilia development TALPID3 is essential for migration and orientation of centrosomes during ciliogenesis TALPID3 is not essential for centriole length TALPID3 is not essential for centriol duplication TALPID3 is not essential for axoneme formation Telencephalic choroid plexus forms in in talpid3-/- chicken embryos Lung development is abnormal in talpid3-/- and phenocopies human ciliopathy syndromes Liver development is abnormal in talpid3-/- and phenocopies human ciliopathy syndromes Inner ear development is abnormal in talpid3-/- and phenocopies human ciliopathy syndromes talpid3-/- embryos have polycystic kidneys talpid3-/- embryos have planar cell polarity type anatomical defects talpid3-/- embryos/cells have intracellular mislocalisation of core planar cell polarity proteins TALPID3 is not epistatic to KIF7 TALPID3 is not epistatic to SMO TALPID3 mutations in human cause Joubert syndrome |
Exploitation Route | TALPID3 may be a target to regulation aberrant Hedgehog signalling (i.e. a significant cause of pancreatic and other cancers) TALPID3 mutations may be used to diagnose Joubert syndrome |
Sectors | Healthcare |
Description | TALPID3 mutations have been found to cause human Joubert Syndrome. Preimplantation diagnosis has been used to slelect unaffected embryos in families with Joubert Syndrome. |
First Year Of Impact | 2015 |
Sector | Healthcare,Pharmaceuticals and Medical Biotechnology |
Impact Types | Policy & public services |
Description | British Society for Developmental Biology Travel Award |
Amount | £400 (GBP) |
Organisation | British Society for Developmental Biology |
Sector | Academic/University |
Country | United Kingdom |
Start | 05/2013 |
End | 06/2014 |
Description | CoB Small Meeting |
Amount | £2,000 (GBP) |
Organisation | Company of Biologists |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 04/2013 |
End | 05/2013 |
Description | Nordic Centrosome and Cilia Network |
Organisation | Nordic Cilia and Centrosome Network |
Country | Global |
Sector | Charity/Non Profit |
PI Contribution | Training partnerships, conferences |
Collaborator Contribution | Training partnerships, conferences |
Impact | Training[ proteomics |
Start Year | 2012 |
Title | Preimplantion diagnosis for Joubert Syndrome |
Description | Families with Joubert syndrome have now selected unaffected embryos through pre-implantation diagnosis used whole exome sequencing to discover mutations in TALPID3 |
Type | Diagnostic Tool - Non-Imaging |
Current Stage Of Development | Initial development |
Year Development Stage Completed | 2016 |
Development Status | Under active development/distribution |
Impact | Detection of TALPID3 mutations in children with Jeune's syndrome |
Description | Doors Open Day Roslin 2013 |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | Yes |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | Sparked interest and questions School students sought out information on STEM careers |
Year(s) Of Engagement Activity | 2013 |
Description | Media Interest (hand development) |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | Yes |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Interview with radio Scotland, coverage article in The Times, The Metro Many people have contacted me regarding interest in becoming a scientist, and learning more about the use of animals in research |
Year(s) Of Engagement Activity | 2014 |
Description | Midlothian Science Festival 2014 |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | Yes |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | increased awareness of use of animals in research, sparked interest in becoming scientist Students said they were open to career in STEM |
Year(s) Of Engagement Activity | 2014 |
Description | National Science and Engineering Competition |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | Yes |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | The participant from the lab went on to take STEM subject at university The student went on to STEM subjects at university |
Year(s) Of Engagement Activity | 2013 |
Description | Nuffield Research Placement |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | Yes |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | Encouraged STEM careers Students went on to take STEM subjects at university |
Year(s) Of Engagement Activity | 2009,2010,2011,2012,2013,2014 |
Description | School visit (Edinburgh) |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Schools |
Results and Impact | Talk, practical, discussion Participants were more open to animal research |
Year(s) Of Engagement Activity | 2013 |
Description | Soapbox Science Edinburgh 2016 |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | Soapbox Science is an informal forum in which female scientists present their work in public using a 'soapbox' format in a public area. The passing public are encouraged to stop and listen and ask questions of the scientists. Social media was used extensively before and after the event. |
Year(s) Of Engagement Activity | 2016 |
URL | https://youtu.be/WEvzU_soOJ4 |
Description | TEDX Talk-"This is a Golden Age of Science- Grab it with Both hands |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | This was an invited TEDX talk which has been view online more than 700 times and was the most highly viewed talk of the TEDX event. The talk covered the impacts of genome sequencing technology and open access publication. Subsequently Dr Davey was invited to attend other public engagement events. |
Year(s) Of Engagement Activity | 2016 |
URL | https://youtu.be/vw7sTgQiltI?list=PL1c4W6FJ_82YOOiJVbZUZlfj2ysg_4aYX |