Gnathostome dental pattern and the evolution of chondrichthyan dentitions

Lead Research Organisation: University of Sheffield
Department Name: Animal and Plant Sciences

Abstract

One of the most significant vertebrate innovations was the evolution of jaws, and on these jaws, teeth arranged into functional dentitions. These dentitions allowed vertebrates to radiate into a number of major groups, including sharks (Chondrichthyes) and bony fishes
(Osteichthyes). Along with these are a variety of fossil groups whose dentitions have been difficult to understand and interpret. Because these fossil groups are closer to the origin of jaws and teeth, it is crucial that we try to better understand how their dentitions evolved.

One way to do this is to study how the dentitions of living groups developed and then apply these observations, as far as possible, to the development of dentitions in fossil taxa. The group Osteichthyes includes well-known fish such as the tuna, salmon, seahorse, coelacanth, lungfish and also tetrapods. Osteichthyan dentitions have been intensely studied in recent years, and we even understand what genes are involved in producing these dentitions. Despite this, even within this group we know nothing about tooth development of more primitive osteichthyans, such as the paddlefish and gar. As well, and perhaps somewhat surprisingly, we know very little about how genes control
development of shark and ray dentitions. The goal of our project is to study development in a broad range of chondrichthyans to find common developmental characters of these dentitions that can be compared to osteichthyan dentitions and in the future, to the more primitive, problematic fossil taxa. Shark teeth are plentiful in the fossil record and features they show are important in identifying different species. Chondrichthyans show a wide variety of dentitions, ranging from large numbers of almost microscopic teeth along the jaw (in filter-feeding sharks such as Rhincodon, the Whale shark), to the pavement-like dentition in rays, to a dentition in which teeth change along the jaw from biting to crushing (Heterodontus, the Port Jackson shark). This diversity has made it difficult to find common patterns of dentition development
that could be used to compare to the problematic fossil vertebrates mentioned above. Sharks and rays also possess other 'tooth-like' structures such as the gill rakers in the filter-feeding basking shark Cetorhinus and 'teeth' along the rostrum of sawfish and sawsharks. Our project will study specimens of a wide range of modern sharks and rays of different growth stages in order to investigate the patterns of tooth development within the jaws and changes in the number and shape of teeth during growth. A number of exceptionally well-preserved fossils will also be studied in order to place the modern forms within a wider context within the Chondrichthyes. Additionally, we will study embryos of a shark, ray and a basal bony fish and will record the genes controlling their tooth development that will allow us to compare underlying mechanisms of the formation of teeth in all vertebrates.

Planned Impact

1) Academic Beneficiaries: see 'Academic Beneficiaries' page.
2) Citizen Scientists: Shark teeth/dentitions have a high degree of interest among amateur collectors/researchers. In the fossil fish collections at the Natural History Museum, London (NHM), shark teeth represent a large portion of donations to the collection, and general enquiries. This is due to the easy availability of relevant shark-bearing geological formations outside London. Our work will be of interest to these citizen scientists as it will provide new characters to help them identify and classify their fossil finds, and illustrate a new aspect to their fossils (ontogeny, or how teeth/dentitions develop through growth) that they may not have considered. In this way, our project will contribute to increasing public awareness and understanding of scientific issues. We will develop an outreach/learning packet that can be modified to meet the needs of these beneficiaries; our team members will use these and each plan to attend one internal (e.g., NHM Nature Live), and two external UK events per year of the grant, to better disseminate our research and increase the impact of our work (see role of 'virtual palaeontology', below). We can also reach citizen scientists via NHM websites such as 'Nature Plus' (http://www.nhm.ac.uk/natureplus/index.jspa) and KEEmu, where we can blog project progress, including strong visuals such as reconstructed CT-scans, histological images, gene expression results, etc.
3) Students (schools) and adult education: These beneficiaries will overlap with 'citizen scientists', but we will modify our outreach events to maximize our impact with these beneficiaries. After initial discussions, we could tailor our outreach packet for individual schools and curricula. For example, we would be more specimen-oriented (Recent shark jaws, histological slides, cleared and stained specimens) for a biology class. For a class interested in the technological side of our research, we could focus on CT-scanning (using images and videos on a portable laptop) and 3-D rapid prototyping (both aspects of outreach and education via 'virtual palaeontology', Rahman et al., 2012. Evo Edu Outreach
DOI 10.1007/s12052-012-0458-2). By disseminating our results in this flexible, adaptive way, we will contribute to the scientific culture in the UK and Europe. Students could also access websites as above.
4) Museums, galleries and aquaria: Given the wide public interest in shark teeth/dentitions, our work will be beneficial to museums and galleries that may be able to incorporate our results into exhibitions or for use in museum talks or public engagement. For example, PI Johanson regularly contributes to NHM public engagement via 'Nature Live', 'Science Uncovered' and 'A-levels' schools programme. Project Partner Sansom has already liaised with the Sea Life Centre in Birmingham such that we will provide them a version of our outreach to use. The Sea Life Centre has a network throughout Europe providing opportunities for outreach in continental Europe as well (targeting facilities in Germany).
5) Staff working on this project: Staff will acquire important skills from either CT-scanning (NHM, KCL), including using the software needed to structurally render this data, or, analysis of gene expression patterns via in situ hybridization. These are highly mature technologies and used in a wide variety of research programs involving both fossil and extant specimens. In this way, our project will train and deliver highly skilled researchers and technicians. Additionally, the PDRA (Sheffield) will travel to the lab of Project Partner Marcus Davis (Kennesaw State University), having the opportunity to develop their skills and interact in a new lab environment. Our timeline suggests these benefits accrue
within the first year with respect to the CT-scanning, and within the first two years for the molecular work.

Publications

10 25 50
 
Description We have recently published key findings on the research - that sharks develop their dentition using a core gene network conserved among all vertebrates. They then utilise this network to regenerate their dentition continuously throughout life. Findings were published in Developmental Biology with a current Altmetric Score of 334.

ADDITION: Further to this we have more recently (2016) produced a manuscript now published in Proceedings of the National Academy of Sciences, (PNAS 2016 December, 113 (51) 14769-14774) where we have discovered that shark tooth regeneration is governed by specialised stem cells that are shared with mouth taste buds - we have therefore uncovered an evolutionary transition from skin teeth to regenerative oral teeth that emerged from the adoption of taste bud stem cells that offered teeth a regenerative function through the activity of the gene Sox2. Therefore, teeth in vertebrates became 'true' teeth i.e. regenerative, through the combined evolution of tooth-like structures developing in taste bud-rich regions of the mouth to enable new regenerative capabilities.
http://www.pnas.org/content/113/51/14769.abstract
https://theconversation.com/shark-study-reveals-taste-buds-were-key-to-evolution-of-teeth-71376
Exploitation Route We will publish another three manuscripts from this specific output - including a stepwise progressive set of data that aim to begin the future research of comparing human tooth formation and lack of regeneration with the shark system (from data obtained during this project). This will be followed by further grant applications to move this science into more translational and public reaching outcomes.
Sectors Aerospace, Defence and Marine,Education,Healthcare,Pharmaceuticals and Medical Biotechnology,Transport

URL https://www.sheffield.ac.uk/news/nr/shark-teeth-human-tooth-loss-study-1.548598
 
Title Culture techniques, NC3Rs and development of methods for Next Generation sequencing. 
Description This method has been developed in an effort to maximise data on the development and regeneration of the dentition, without increasing the number of animals used/sacrificed. We use in situ hybridisation techniques to identify the genes expressed in the developing dentition. Initially, we replaced whole animal use for a single gene (whole mount) analysis with paraffin sections; where multiple sections from the same animal can be used to identify the expression of several genes - thus reducing the number of animals used in the experiments. Following this method shift, we have begun tissue culturing techniques allowing the understanding of gene function with out the need to manipulate living animals - the a replacement/reduction (NC3Rs) technique that we have developed. Other developments that have arisen from this research is a cell culture method, which would allow the maintenance of cell lines to manipulate rather than live animals to understand function of genes responsible for tooth regeneration and development. These techniques will also combine to develop a set of follow up experiments for next generation sequencing (RNAseq) of specific cells types (this will be the basis of a follow grant proposal). 
Type Of Material Model of mechanisms or symptoms - in vitro 
Provided To Others? No  
Impact Notable impacts of these experimental advances are that this pushes the boundaries of non-model organismal experimentation, without increasing the number of animals used. These experiments will push the research/field into a new understanding of the evolution, development, and genetics of dentitions. These results/impacts are not specific to just sharks and fishes but have important implications across all vertebrates, including humans. This link between the research we are undertaking and the human condition will be the major research impact. 
 
Title Scratchpad Research Database 
Description We have collectively (operated by Dr. Zerina Johanson, and hosted by the Natural History Museum) developed a web-based database (Scratchpad) to update and post research progress. 
Type Of Material Database/Collection of data 
Provided To Others? No  
Impact The major impact of this database is the general public access and transparency of this NERC-funded research for public and scientific use. 
URL http://chondrichthyes.myspecies.info/
 
Description BBC Radio 3 'Free Thinking' guest expert. 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Requested to be a guest expert on the BBC Radio 3 show 'Free Thinking' to discuss sharks and our research in the context of a debate on the impact of Jaws the book and movie. Interaction with the Radio hosts and producers will lead to other radio productions, and other impacts include request for information, interest in research in a wider context.
Year(s) Of Engagement Activity 2015
URL http://www.bbc.co.uk/programmes/b060zryf
 
Description BBC Radio and TV 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Following the publication of the recent Developmental Biology paper BBC Radio (5 Live, World Service, BBC Sheffield, and BBC Scotland) and BBC News website and BBC Look North (TV) invited PI Fraser to be interviewed to discuss the research on shark teeth and regeneration. The wider implications were huge and international media recognition for this was the result. Twitter users discussed the media output of this research and many social network discussions were sparked including other media entries including a Daily Mail Newspaper article.
Year(s) Of Engagement Activity 2016
URL http://www.bbc.co.uk/news/uk-england-south-yorkshire-35599025
 
Description General Media Article written for 'The Conversation' - Shark study reveals taste buds were key to evolution of teeth. 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact This article was written for The Conversation - online - to engage a wider audience with our research on tooth regeneration and evolution. This targeted an international audience, sparking engagement and further requests for participation and further collaboration.
Year(s) Of Engagement Activity 2017
URL https://theconversation.com/shark-study-reveals-taste-buds-were-key-to-evolution-of-teeth-71376
 
Description General Media Article written for 'The Conversation'- How sharks could help us regrow our own human teeth?. 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact A general Science article written to explain our latest research highlights and a proposal for the future of the research, in the context of human health. This led to another article requested by 'The Conversation' based on new developments from the grant.
Year(s) Of Engagement Activity 2016
URL https://theconversation.com/how-sharks-could-help-us-regrow-our-own-human-teeth-63531
 
Description Interview BBC Radio 4 'In Our Time' programme. 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Invitation and broadcast on the BBC Radio 4 programme 'In our Time' hosted by Melvyn Bragg. PI Fraser was on of the experts.
Year(s) Of Engagement Activity 2019
URL https://www.bbc.co.uk/programmes/m0003zbg
 
Description Interview and segment for a National Geographic Channel documentary 'Pushing the Boundaries of Dentistry'. 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Invited to record an interview and a filmed segment was the focus of my research associated with the NERC grant results. The documentary was made for and presented by National Geographic Channel (USA) and aired in January 2019.
Year(s) Of Engagement Activity 2019
URL https://www.nationalgeographic.co.uk/science-and-technology/2018/11/pushing-boundaries-technology-0?...
 
Description Natural History Society Outreach 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Undergraduate students
Results and Impact Natural History Society invited talk helps to bring the research of the grant to wider student and public audience. Interactive talk enables members of the public to engage with the research outcomes and to ask questions and discuss the wider implications of the science.
Year(s) Of Engagement Activity 2016
 
Description Outreach - University of Sheffield, Festival of the Mind - Public engagement. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact A number of discussions resulted from the talk/engagement activity, and further discussions/questions with members of the public continued afterwards. This discussion continued on social media - where further connections were forged.

From the presentation at the Festival of the Mind - a number of parents asked if their children could visit my lab and if the school could contact my Department about further outreach sessions. Our Department currently runs a number of school group sessions as an outreach activity, with which I will be involved with in the coming year(s).
Year(s) Of Engagement Activity 2014
URL http://festivalofthemind.group.shef.ac.uk/cabinet-curiosities-alfred-denny-museum/
 
Description Researcher's Night, University of Sheffield 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact This was a talk and demonstration of the research conducted in my lab, directly related to my NERC funded research. This activity sparked discussion, debate and stimulated thinking in a wide audience, from school children to adults of a varied background. Many groups participated in debate and discussions while in the demonstration.

Following the researchers night demonstration i received feedback from several members of the public, including parent's of children who were inspired by the evening - now showing an increased interest in biology/zoology as a result.

Following and as a result of the researchers night demonstration I was asked to give the Christmas Lecture for the Science Brainwaves outreach event.
Year(s) Of Engagement Activity 2014
 
Description TV segment on Discovery Channel Canada for Shark Week 2016 - Programme 'Daily Planet' 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact TV segment was viewed by the general public in Canada and wider audiences. This TV spot has led to the request for further media productions notably PI FRASER is now producing and presenting a new TV documentary for Discovery Channel International in 2017 focused on the science of shark teeth called 'Tooth Hunters'. To be aired in August 2017.
Year(s) Of Engagement Activity 2016