Roslin Institute Research Articles

Adjusting to precarity: how and why the Roslin Institute forged a leading role for itself in international networks of pig genomics research.

From the 1980s onwards, the Roslin Institute and its predecessor organizations faced budget cuts, organizational upheaval and considerable insecurity. Over the next few decades, it was transformed by the introduction of molecular biology and transgenic research, but remained a hub of animal geneticists conducting research aimed at the livestock-breeding industry. This paper explores how these animal geneticists embraced genomics in response to the many-faceted precarity that the Roslin Institute faced, establishing it as a global centre for pig genomics research through forging and leading the Pig Gene Mapping Project (PiGMaP); developing and hosting resources, such as a database for genetic linkage data; and producing associated statistical and software tools to analyse the data. The Roslin Institute leveraged these resources to play a key role in further international collaborations as a hedge against precarity. This adoption of genomics was strategically useful, as it took advantage of policy shifts at the national and European levels towards funding research with biotechnological potential. As genomics constitutes a set of infrastructures and resources with manifold uses, the development of capabilities in this domain also helped Roslin to diversify as a response to precarity.

When does innovation go too far?

SHOULD innovative veterinary procedures and practice be more regulated? This question was considered during a debate at the recent Animal Welfare Foundation discussion forum. One problem encountered in trying to answer the question is the number of variables that must be considered. First, what can be done under current regulations? Experimental procedures and research are licensed under the Animals (Scientific Procedures) Act 1986 (ASPA), and this can include research carried out on farms and in veterinary practices (for more details on the Act see In Practice, November 2020, vol 42, pp 488–496). However, most vets will be operating under the Veterinary Surgeons Act 1966 (VSA), conducting ‘recognised veterinary practice’. According to the RCVS: ‘The clinical investigation and management of the health or welfare of animals is generally considered to be recognised veterinary practice when it involves an intervention which is of direct benefit to the animal or its immediate peer group.‘ The RCVS gives more advice in the supporting guidance to the code of professional conduct on what actions fall within the ASPA, the VSA or the Veterinary Medicines Regulations 2011, but it is impossible to cover all situations, as the RCVS itself acknowledges. So another variable is what is considered innovative? And if something is considered to be new, should it be regarded as recognised veterinary practice? And who judges this? In many areas, those speaking on both sides of the debate during the discussion forum – Eddie Clutton, a specialist anaesthetist at Edinburgh university and the Roslin Institute, and Karen Humm, a critical care specialist at the Royal Veterinary College – were in agreement. For example, that euthanasia should be part of the discussion for cases being considered for innovative treatment; that humane endpoints should be set so there is no clinical ‘creep’ in outcome that becomes detrimental to welfare; and that certain areas where the evidence base is strong, such as using the medicines' cascade, are suitable for self-regulation. Another area on which there was agreement was that more uncertain ‘heroic’ treatment should be subject to greater ethical overview. Clutton argued that current arrangements were open to potential conflicts of interest and new arrangements should be put in place to create an independent mandatory ‘innovation panel’ to assess the ethics of a proposed treatment and whether it offered enough potential benefit to an animal (or its ‘immediate peer group’). There would need to be the ability to assess treatment after the event if it was conducted in an emergency. Procedures and treatments conducted ‘should not be at the whim of individuals’, said Clutton, and this would go some way to ensuring unproven methods had better scrutiny. Although there will be vets who push the limits for their own promotion or challenge, or because their employer rewards ‘firsts’, the biggest push for unethical treatment came from clients, according to Humm. Often, they had heard or read of a treatment or procedure offered in other countries, and, she said, in such situations, the most essential skill was the ability to explain to these clients why that option was not on offer, rather than the technical skill required to perform it. “Any change to improve regulation would likely be met with enthusiasm Independent ethical overview makes sense. It means new methods can be introduced but acknowledges the need to assess them for an animal's best interests. No one is arguing that veterinary practice should not develop and improve though innovation, but rather, by taking this course of action, evidence will be gathered on how effective new treatments are and ensure that, as a reason, ‘because we can’ is taken out of the equation. BVA grants editorial freedom to the Editor of Vet Record. The views expressed in the journal are those of the authors and may not necessarily comply with BVA policy

In conversation with Christine Watson.

Christine J. Watson is Professor of Cell and Cancer Biology at the University of Cambridge. Christine obtained her Bachelor's (honors) degree in Biochemistry at the University of Glasgow in 1975 and, after a soujourn in Glauco Tocchini-Valentini's lab at the Institute of Cell Biology, Consiglio Nazionale delle Ricerche in Rome, she undertook a PhD in Molecular Genetics at Imperial College London. During her PhD, she looked at differences in gene expression between differentiated and undifferentiated embryonal carcinoma stem cells, inspiring an early interest in gene expression and cell fate determination. Between 1986 and 1992, Christine undertook three postdoctoral research positions that took her from London back to Scotland, where she was first introduced to mammary gland biology through her work with John Clark at the Roslin Institute in Edinburgh. During her time in the Clark lab, Christine identified a factor - later shown to be STAT5 - that binds to the promoter of the milk protein gene β-lactoglobulin. This prompted further work identifying the key role played by the STAT family of transcription factors in mammary gland development. Shortly afterwards, Christine became a group leader at the Roslin Institute and later relocated to the University of Edinburgh to collaborate with Andrew Wyllie. This led to her recruitment to the University of Cambridge in 1998, where she has remained to date. Over the last two decades, the Watson lab has focused on elucidating the mechanisms underlying lineage commitment of mammary stem and progenitor cells and the regulation of cell death in involuting mammary gland. In this interview, Christine discusses her research highlights and provides a glimpse into her personal interests, as she moves towards retirement.

Update on possible animal sources for COVID-19 in humans.

XenotransplantationVolume 27, Issue 3 e12621 COMMENTARY Update on possible animal sources for COVID-19 in humans Tanja Opriessnig, Corresponding Author Tanja Opriessnig Tanja.opriessnig@roslin.ed.ac.uk tanjaopr@iatate.edu orcid.org/0000-0001-9642-0904 The Roslin Institute and The Royal (Dick) School of Veterinary Studies, University of Edinburgh, Midlothian, UK Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA, USA Correspondence Tanja Opriessnig, The Roslin Institute and The Royal (Dick) School of Veterinary Studies, University of Edinburgh, Midlothian, UK. Emails: Tanja.Opriessnig@roslin.ed.a.ac.uk, tanja.opriessnig@roslin.ed.ac.ukSearch for more papers by this authorYao-Wei Huang, Yao-Wei Huang Institute of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, ChinaSearch for more papers by this author Tanja Opriessnig, Corresponding Author Tanja Opriessnig Tanja.opriessnig@roslin.ed.ac.uk tanjaopr@iatate.edu orcid.org/0000-0001-9642-0904 The Roslin Institute and The Royal (Dick) School of Veterinary Studies, University of Edinburgh, Midlothian, UK Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA, USA Correspondence Tanja Opriessnig, The Roslin Institute and The Royal (Dick) School of Veterinary Studies, University of Edinburgh, Midlothian, UK. Emails: Tanja.Opriessnig@roslin.ed.a.ac.uk, tanja.opriessnig@roslin.ed.ac.ukSearch for more papers by this authorYao-Wei Huang, Yao-Wei Huang Institute of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, ChinaSearch for more papers by this author First published: 17 June 2020 https://doi.org/10.1111/xen.12621Citations: 7Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume27, Issue3May/June 2020e12621 RelatedInformation

First person – Claire L. Wood

ABSTRACTFirst Person is a series of interviews with the first authors of a selection of papers published in Disease Models & Mechanisms (DMM), helping early-career researchers promote themselves alongside their papers. Claire Wood is first author on ‘A comparison of the bone and growth phenotype of mdx, mdx:Cmah−/− and mdx:Utrn+/− murine models with the C57BL/10 wild-type mouse’, published in DMM. Claire conducted the research described in this article while an MRC Clinical Research Training Fellow in Colin Farquharson's lab at Roslin Institute, University of Edinburgh, UK. She is now an academic clinical lecturer in the lab of Tim Cheetham and Simon Pearce at the Institute of Genetic Medicine, Newcastle University, UK, investigating bone and growth in children.

A Review Article on Study of Cloning

DNA cloning, that can be used to discover new ways of warding off diseases and ailments is good and by no means harmful. A group of genetically identical cells produced by mitotic division from an original cell. The production of genetically identical animals by ‘embryo splitting ’. This can occur naturally at the two cell stage to give identical twins. The type of cloning which we will focus on here is called Nuclear Transfer. Cloning requires specialized microsurgery tools. There are two ways to make an exact genetic copy of an organism in a lab artificial embryo twinning and somatic cell nuclear transfer. In 1996, the Roslin Institute, England cloned “Dolly, ” a sheep to demonstrate that somatic cells were able to be used as donor cells for the procedure, prior to this blastoceles (stem cells) were used in nuclear transfer technology. Around 2004–2005, in South Korean scientists falsely claimed to have used somatic cell nuclear transfer to create embryonic stem cell lines, the scientific community demanded much stronger evidence that the procedure had actually been successful. Cloned buffalo gave birth to a baby buffalo in India. In 2007 Primate embryonic stem cells created by somatic cell nuclear transfer. A DNA from an organism is transferred to a self-replicating genetic element such as a bacterial plasmid. Reproductive cloning is the basis of most controversial debates regarding the Genetic Revolution. The California Advising Committee on Human Cloning found that: Attempts at cloning certain species such as monkeys, chickens, horses and dogs have been unsuccessful.

Monkey see, Humans Won't Do-the misguided reaction to the first cloning of primates.

At the end of January, a paper in Cell announced the first successful cloning of primates [1]. Researchers at the Chinese Academy of Sciences in Shanghai reported that they had successfully created two female macaque clones about 8 weeks old. The Chinese researchers made clear their goal in undertaking the experiment was to find a way to create genetically uniform primates for use in research. Nonetheless, the subsequent conversation about the cloning achievement focused on the feasibility of doing so in humans—indeed, most press accounts and ethical commentary immediately jumped into speculation about whether human cloning would soon follow. The media obsession with the prospect of human cloning is not new. It is an extension of the misplaced ethical concerns that accompanied media reports and commentary surrounding the announcement in 1997 of the world's first cloned mammal, Dolly, the sheep at the Roslin Institute in Midlothian, Scotland [2]. Not only were those concerns unfounded [3], they sadly continue to shape ethical discourse about cloning to the present day. Many news accounts reported that the Chinese team had cloned primates using the same somatic …

Cuts and the cutting edge: British science funding and the making of animal biotechnology in 1980s Edinburgh.

The Animal Breeding Research Organisation in Edinburgh (ABRO, founded in 1945) was a direct ancestor of the Roslin Institute, celebrated for the cloning of Dolly the sheep. After a period of sustained growth as an institute of the Agricultural Research Council (ARC), ABRO was to lose most of its funding in 1981. This decision has been absorbed into the narrative of the Thatcherite attack on science, but in this article I show that the choice to restructure ABRO pre-dated major government cuts to agricultural research, and stemmed from the ARC's wish to prioritize biotechnology in its portfolio. ABRO's management embraced this wish and campaigned against the cuts based on a promise of biotechnological innovation, shifting its focus from farm animal genetics to the production of recombinant pharmaceuticals in sheep milk. By tracing interaction between government policies, research council agendas and local strategies, I show how novel research programmes such as genetic modification could act as a lifeline for struggling institutions.

Dolly and Alice.

The opinion of the United States Court of Appeals for the Federal Circuit, In re Roslin Institute, rejecting patent claims to mammals cloned from somatic cells, was rendered about a month before the United States Supreme Court's decision in Alice Corp. v. CLS Bank International. The Alice opinion explicitly sets out the standard for determining whether an invention falls within statutory patentable subject matter. Thus one is thus left to wonder what the Roslin opinion might have looked like had it been decided only a few weeks later, after the Alice decision was published, with the benefit of the Supreme Court's further direction on patentable subject matter. In this essay I explore whether in hindsight the Alice standard might have dictated a different outcome in Roslin, suggesting how the two-part test articulated by the Supreme Court in Alice might apply to a ‘products of nature’ analysis for cloned mammals. Drawing on that analysis, I then use the Roslin case as a vehicle to highlight certain issues with the Supreme Court's current subject matter jurisprudence as applied to biotechnology. By juxtaposing Dolly with Alice, it becomes clear that the Supreme Court has revivified a number of dormant biotechnology patent problems in the guise of subject matter analysis.

The BBSRC DRINC programme - Pre-competitive research and industry partnerships in diet and health

Nutrition BulletinVolume 40, Issue 2 p. 130-133 News from the Research Councils The BBSRC DRINC programme – Pre-competitive research and industry partnerships in diet and health B. Hanley, Corresponding Author B. Hanley Knowledge Transfer Network, UK Correspondence: Prof Bryan Hanley, Specialist, Food, Knowledge Transfer Network, The Roslin Institute, Easter Bush, Midlothian, EH25 9RG. E-mail: bryan.hanley@ktn-uk.orgSearch for more papers by this authorS. Broadway, S. Broadway Knowledge Transfer Network, UKSearch for more papers by this authorJ. Brookman, J. Brookman Knowledge Transfer Network, UKSearch for more papers by this author B. Hanley, Corresponding Author B. Hanley Knowledge Transfer Network, UK Correspondence: Prof Bryan Hanley, Specialist, Food, Knowledge Transfer Network, The Roslin Institute, Easter Bush, Midlothian, EH25 9RG. E-mail: bryan.hanley@ktn-uk.orgSearch for more papers by this authorS. Broadway, S. Broadway Knowledge Transfer Network, UKSearch for more papers by this authorJ. Brookman, J. Brookman Knowledge Transfer Network, UKSearch for more papers by this author First published: 19 May 2015 https://doi.org/10.1111/nbu.12142Citations: 2Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume40, Issue2June 2015Pages 130-133 RelatedInformation

PARV4: an emerging tetraparvovirus.

1 Nuffield Department of Medicine, Peter Medawar Building for Pathogen Research, Oxford, United Kingdom, 2 Department of Infectious Diseases and Microbiology, Oxford University Hospitals NHS Trust, John Radcliffe Hospital, Oxford, United Kingdom, 3 Department of Paediatrics, Peter Medawar Building for Pathogen Research, Oxford, United Kingdom, 4 The Roslin Institute, The University of Edinburgh, Easter Bush, Midlothian, Scotland, United Kingdom, 5 NIHR Biomedical Research Centre, John Radcliffe Hospital, Headington, Oxford, United Kingdom

From germ cell preservation to regenerative medicine: an exciting research career in biotechnology.

Collection, manipulation, assessment, and storage of mammalian gametes, embryos, and stem cells are providing important opportunities in agriculture, research, and medicine. Semen and embryo freezing in livestock are used in breeding schemes, especially in cattle and for international trade, with no risk of spreading disease. In human medicine, they are used in treatment of infertility. Usually, knowledge gained in one species is applicable in the others. In one exception, some ruminant embryos cultured according to protocols used in human in vitro fertilization become unusually large offspring. This is due to disturbances in expression of imprinted genes. The nuclear transfer procedure developed at the Roslin Institute is being used to make genetic modifications in livestock to either direct production of biomedical proteins, create animal models of human disease, or enhance animal health and productivity. Human pluripotent cells are being used in Edinburgh to identify drugs to treat degenerative diseases.

Technical advances in the generation of transgenic animals and in their applications. Nantes, France, June 7th 2013

On June 7 2013 was held in Nantes, France, the international meeting ‘‘Technical advances in the generation of transgenic animals and in their applications’’ (www.trm.nantes.inserm.fr/index.php). This biennial meeting is the fourth one of its kind to be organized by the Transgenic Rats Nantes facility (http:// www.tgr.nantes.inserm.fr/). The meeting was supported by private companies (Charles River, genOway, Lonza, Sigma-Aldrich and TaconicArtemis GmbH) and by public institutions (University of Nantes, INSERM UMR 1064, SFR Francois Bonamy, CNRS, Region Pays de la Loire, Biogenouest, IBiSA and IHUCESTI). This meeting received support and co-sponsorhip from the International Society for Transgenic Technology. Around 100 participants, from France but also from different European countries and USA, attended the meeting and registration prices were kept as low as possible to facilitate the participation of students. The meeting aimed to provide an update on recent technical developments in the generation of transgenic animals and in their applications. It was intended for Master, PhD and medical students as well as for researchers with a background in molecular biology and genetics as an introduction to future work in these rapidly developing areas of research. Bruce Whitelaw (The Roslin Institute, University of Edinburgh, UK) gave an overview on the advances and new trends in the generation of genetically engineered animals (GE) particularly in livestock. GE animals have existed for over 30 years (Clark and Whitelaw 2003). Throughout this time the tools available for transferring genetic change into the genome have evolved to the current position where we have both efficient and precise methods. There are two general approaches to GE; either delivery to the zygote or to introducing engineered cells into the early embryo. Both approaches can now be combined with the exciting new editor technologies to enable precise genome editing which in some cases does not involve the incorporation of a transgene. Depending on the species working with—that include rodents, rabbits, fish, birds and livestock—the different GE strategies offer advantages. For example, transgene delivery by lentivirus vectors is attractive for livestock and birds but of lesser importance for mice; in livestock where gestation and breeding times are long, this method dramatically reduces the time to proof-of-concept for a S. Menoret L. Tesson S. Remy R. Thinard C. Usal L.-H. Ouisse V. Thepenier I. Anegon Transgenic Rats Nantes, INSERM, Centre National de Recherche Scientifique, 44093 Nantes, France

Development of a monoclonal antibody against the canine CCR2 as a new targeted anticancer therapy in the scope of comparative oncology

Event Abstract Back to Event Development of a monoclonal antibody against the canine CCR2 as a new targeted anticancer therapy in the scope of comparative oncology Teresa P. Raposo1*, David J. Argyle2, Breno C. Beirao2, Justina Prada1, Isabel Pires1 and Felisbina L. Queiroga1 1 University of Trás-os-Montes and Alto Douro, Department of Veterinary Sciences, Portugal 2 University of Edinburgh, The Royal (Dick) School of Veterinary Studies and The Roslin Institute, United Kingdom Introduction: In this project, it is proposed that the clinical application of a monoclonal antibody against the receptor of the Monocyte Chemotactic Protein-1 (MAb anti-CCR2), would hinder cancer progression, inhibiting macrophages recruitment. Canine mammary tumour (CMT) will be used as a model of human breast cancer, in face of molecular, histologic and clinical similarities of the disease in both species. Herein, the initial stages of the production of a MAb anti-CCR2 are described, aiming the future clinical application. Material and Methods: RNA was extracted from canine bone marrow, CCR2 cDNA was synthesized by RT-PCR, cloned into a plasmid vector, and then confirmed through DNA sequencing. TransformedBL21 DE3 E.coli were used as a system of bacterial expression. The bacterial lysates were purified through an affinity chromatography column, to isolate the CCR2 protein which was subsequently verified through mass spectrometry. Results: The purity of the isolated protein was confirmed by mass spectrometry analysis and it is now suitable for the mice immunisation. Throughout the purification process, the inclusion of urea in the lysis, wash and elution buffers was crucial to withdraw the most of the protein from the bacterial lysates. Conclusion: Hitherto, the antigen production stage was concluded, obtaining a pure form of CCR2, ready for the immunisation of mice.Hence, it is now possible to progress to the following phases in the development of a Mab anti-CCR2 using hybridoma technology. Afterwards, an experimental therapy delivering anti-CCR2 monoclonal antibody to the tumour site will be applied to dogs with advanced CMT. Acknowledgements Ministry of Science and Education, Portugal: QREN - POPH - FCT Project: SFRH / BD / 79158 / 2011 Keywords: tumour-associated macrophages, TAMs, canine cancer model, CCR2 antibody, MAb technology Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Translational immunology and immune intervention Citation: Raposo TP, Argyle DJ, Beirao BC, Prada J, Pires I and Queiroga FL (2013). Development of a monoclonal antibody against the canine CCR2 as a new targeted anticancer therapy in the scope of comparative oncology. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00269 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 12 Mar 2013; Published Online: 22 Aug 2013. * Correspondence: Miss. Teresa P Raposo, University of Trás-os-Montes and Alto Douro, Department of Veterinary Sciences, Vila Real, Portugal, rapteresa@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Teresa P Raposo David J Argyle Breno C Beirao Justina Prada Isabel Pires Felisbina L Queiroga Google Teresa P Raposo David J Argyle Breno C Beirao Justina Prada Isabel Pires Felisbina L Queiroga Google Scholar Teresa P Raposo David J Argyle Breno C Beirao Justina Prada Isabel Pires Felisbina L Queiroga PubMed Teresa P Raposo David J Argyle Breno C Beirao Justina Prada Isabel Pires Felisbina L Queiroga Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

DISEASE RESISTANCE AND OTHER APPLICATIONS OF TRANSGENESIS IN THE CHICKEN

Genetic modification of the chicken in terms of gene addition is now robust and efficient. Transgenes can be introduced by injection of lentiviral vectors into chick embryos or by transfection of transposon vectors into embryos or primordial germ cells in vitro. Lentiviral vectors are limited in the size of transgene they can incorporate but we have generated several different transgenic lines using HIV-derived vectors and have observed high levels of transgene expression and tissue-specific expression using regulatory sequences from several genes. M. McGrew (The Roslin Institute) has established primordial germ cell lines and effective methods for transfection with piggyBac and Tol2 transposon vectors. The primordial cells are injected into chick embryos where they populate the developing gonads and contribute to the germline in mature birds. The availability of primordial germ cell lines will also form the basis of using artificial site-specific nucleases for gene knockout and potentially gene targeting in the chicken. These technologies facilitate the application of transgenesis in the chicken for basic research and for potential applications in poultry breeding. The chick embryo is an invaluable model for studying vertebrate development as the embryos can be accessed in ovo or in culture at the earlier stages of development. Embryos can be transfected with transgenes by electroporation and manipulated to study many aspects of development. We are developing transgenic chickens in which fluorescent protein reporters are expressed either ubiquitously or in targeted cell types. These form the basis of novel tools for increasing the value of the chick embryo in studying development. We provide fertile eggs from these lines to other research groups and are investigating the development of macrophages using a macrophage-targeted reporter. The potential for the use of genetic modification to be used in poultry breeding can now be explored. Commercial poultry production is challenged by several major pathogens including avian influenza. Flocks can be protected by good biosecurity measures and/or vaccination but vaccination is not always effective. It may be possible to add novel genes to the chicken genome targeting avian influenza virus replication. We are developing this approach (with L. Tiley, Cambridge University) and have generated transgenic chickens that do not transmit avian influenza when directly infected with H5N1 virus.

An Equine Biobank

Laura Corbin of The Roslin Institute, University of Edinburgh, discusses the work she has carried out on behalf of the British Equestrian Federation

Innovation and Creation

Innovation and Creation

Discordance between cortical and trabecular bone phenotype highlights the role of local versus circulating IGF-1 in the SOCS2 null mouse

Event Abstract Back to Event Discordance between cortical and trabecular bone phenotype highlights the role of local versus circulating IGF-1 in the SOCS2 null mouse R Dobie1*, C Huesa1, RJ Van'T Hof2, VE Macrae1, SF Ahmed3 and C Farquharson1 1 University of Edinburgh, Roslin Institute & R(D)SVS, United Kingdom 2 University of Edinburgh, Centre for Molecular Medicine, United Kingdom 3 University of Glasgow, Bone and Endocrine Research Group, United Kingdom Suppressor of Cytokine Signalling 2 (SOCS2) tightly regulates postnatal growth through its actions on the GH/IGF-1 pathway. SOCS2 knockout mice do not have raised circulating IGF-1 and are characterised by enhanced body size and bone length. However the trabecular architecture and cortical geometry of the bones have yet to be fully investigated. Further to this, the bone mechanical properties remain undefined. In order to better appreciate the role of SOCS2 in juvenile and adult male and female mice, we have conducted an in depth analysis of the bone phenotype. {BR}Micro-CT was carried out on the tibia of wild-type (WT) and SOCS2 knockout (KO) mice, at 6-weeks (juvenile) and 4-months (adult) of age. Analysis enabled detailed comparisons to be made between the trabecular and cortical macroarchitecture, and allowed for the calculation of the mineral density of the bone tissue. 3-point-bending was also completed to investigate the mechanical properties. {BR}A significant increase in body weight was observed in all KO mice (P<0.05). All KO mice had increased trabecular BV/TV compared to their age and sex matched WT controls which is a likely consequence of increased trabecular thickness (P<0.05). All KO mice had increased tibial periosteal surface area (P<0.05), however the cortex was thicker only in juvenile and adult KO male mice. The thicker cortex of the male KO mice tibia was associated with increased work to failure and maximum load. At 6 weeks of age there was no difference in the mineral density of the bone tissue in either gender, however at 4 months both male and female KO mice had a significant decrease of mineral density in the cortical bone (P<0.05).{BR}The increased bone size and mass are consistent with the known anabolic effects of local, skeletal, IGF-1 and these effects are age and sex specific. The fall in mineral density of cortical bone reflects the importance of circulating IGF-1 on cortical bone development. The SOCS2 KO mouse represents a valid model for studying the effects of GH and IGF-1 on bone. Keywords: Bones, Bone Research Conference: 2011 joint meeting of the Bone Research Society & the British Orthopaedic Research Society, Cambridge, United Kingdom, 27 Jun - 29 Jun, 2011. Presentation Type: Oral Topic: Abstracts Citation: Dobie R, Huesa C, Van'T Hof R, Macrae V, Ahmed S and Farquharson C (2011). Discordance between cortical and trabecular bone phenotype highlights the role of local versus circulating IGF-1 in the SOCS2 null mouse. Front. Endocrinol. Conference Abstract: 2011 joint meeting of the Bone Research Society & the British Orthopaedic Research Society. doi: 10.3389/conf.fendo.2011.02.00015 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 30 Sep 2011; Published Online: 30 Sep 2011. * Correspondence: Mr. R Dobie, University of Edinburgh, Roslin Institute & R(D)SVS, United Kingdom, ross.dobie@roslin.ed.ac.uk Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers R Dobie C Huesa RJ Van'T Hof VE Macrae SF Ahmed C Farquharson Google R Dobie C Huesa RJ Van'T Hof VE Macrae SF Ahmed C Farquharson Google Scholar R Dobie C Huesa RJ Van'T Hof VE Macrae SF Ahmed C Farquharson PubMed R Dobie C Huesa RJ Van'T Hof VE Macrae SF Ahmed C Farquharson Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Lack of nucleotide pyrophosphatase phosphodiesterase1 activity in ENPP1-/- mice results in altered bone development and pathological mineralisation of soft tissue

Event Abstract Back to Event Lack of nucleotide pyrophosphatase phosphodiesterase1 activity in ENPP1-/- mice results in altered bone development and pathological mineralisation of soft tissue NCW Mackenzie1*, D Zhu1, EM Milne1, JL Millan2, C Farquharson1 and VE Macrae1 1 University of Edinburgh, the Roslin Institute and Royal (Dick) School of Veterinary Studies, United Kingdom 2 Sanford-Burnam Medical Research Institute, United States Nucleotide pyrophosphatase phosphodiesterase 1 (NPP1) is required to convert extracellular ATP into inorganic pyrophosphate (PPi), a crucial negative regulator of hydroxyapatite (HA) crystal formation. NPP1, together with bone alkaline phosphatase, is responsible for maintaining the balance between PPi and inorganic phosphate (Pi) and thereby regulating the rate of HA formation in the developing skeleton. In this present study a detailed phenotypic assessment of a mouse model lacking NPP1 (Enpp1-/-) was carried out to further the understanding of its role in skeletal and soft tissue mineralisation. Histopathological assessment of Enpp1-/- mice at 22 weeks of age revealed calcification in the aorta and kidney and ectopic cartilage formation in the joints and spine. Radiographic assessment of the hind limb showed evidence of hyper-mineralisation in the talocrural joint and hypo-mineralisation in the femur and tibia. Comprehensive microCT analysis of the tibia and femur examined the effects of Enpp1 ablation on trabecular architecture and bone geometry at 6 and 22 weeks of age in both male and female mice. Trabecular number, trabecular percentage bone volume, structure model index, trabecular and cortical thickness were significantly reduced (P<0.05) in tibiae and femurs from the Enpp1-/- mice compared to wild-type controls. This decreased bone mass was reflected by a significant reduction in maximum bone stiffness in Enpp1-/- tibiae and femurs from 22 week old mice (P<0.05) as determined by three-point bending. Circulating phosphate and calcium plasma levels were also reduced (P<0.05) in the Enpp1-/- null mice. Plasma levels of osteocalcin, a marker of bone formation, were significantly decreased at 6 weeks of age (P<0.05) in Enpp1-/- mice, with no differences noted at 22 weeks of age. Plasma levels of CTx (RatlapsTM ), a marker of bone resorption, and the phosphaturic hormone FGF-23 were significantly increased in the Enpp1-/- mice at 22 weeks of age (P<0.05). These results demonstrate that Enpp1-/- mice are characterised by severe disruption to the architecture and mineralisation of long-bones, dysregulation of calcium/phosphate homeostasis and hypercalcification in soft tissues. In summary, NPP1 is essential for normal bone development and the control of physiological bone mineralisation and pathological mineralisation in soft tissues. Keywords: Bones, Bone Research Conference: 2011 joint meeting of the Bone Research Society & the British Orthopaedic Research Society, Cambridge, United Kingdom, 27 Jun - 29 Jun, 2011. Presentation Type: Oral Topic: Abstracts Citation: Mackenzie N, Zhu D, Milne E, Millan J, Farquharson C and Macrae V (2011). Lack of nucleotide pyrophosphatase phosphodiesterase1 activity in ENPP1-/- mice results in altered bone development and pathological mineralisation of soft tissue. Front. Endocrinol. Conference Abstract: 2011 joint meeting of the Bone Research Society & the British Orthopaedic Research Society. doi: 10.3389/conf.fendo.2011.02.00041 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 30 Sep 2011; Published Online: 30 Sep 2011. * Correspondence: Dr. NCW Mackenzie, University of Edinburgh, the Roslin Institute and Royal (Dick) School of Veterinary Studies, Roslin, United Kingdom, neil.mackenzie@roslin.ed.ac.uk Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers NCW Mackenzie D Zhu EM Milne JL Millan C Farquharson VE Macrae Google NCW Mackenzie D Zhu EM Milne JL Millan C Farquharson VE Macrae Google Scholar NCW Mackenzie D Zhu EM Milne JL Millan C Farquharson VE Macrae PubMed NCW Mackenzie D Zhu EM Milne JL Millan C Farquharson VE Macrae Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

MEPE regulates growth plate mineralisation through its cleavage to the ASARM peptide

Event Abstract Back to Event MEPE regulates growth plate mineralisation through its cleavage to the ASARM peptide KA Staines1*, VE Macrae1 and C Farquharson1 1 the University of Edinburgh, the Roslin Institute and Royal (Dick) School of Veterinary Studies, United Kingdom MEPE is mainly expressed in mineralising tissues, and has been implicated in osteoblast matrix mineralisation directly by releasing an ASARM peptide. Although the growth plates of MEPE transgenic mice display severe morphological disruption, the expression and function of MEPE in growth plate mineralisation remains largely undefined. {BR}Proximal tibiae from 3-week-old wild-type mice were analysed for Mepe expression by in situ hybridisation and growth plate microdissection. Gene expression by the ATDC5 chondrogenic cell line was examined by RT-qPCR over a 20-day culture period under calcifying conditions. 20µM phosphorylated (pASARM) and non-phosphorylated ASARM (npASARM) peptides were added to ATDC5 cultures, and to 17-day-old and 15-day-old embryonic metatarsal explants. {BR}Mepe expression was abundant throughout the growth plate as shown by in situ hybridisation. Microdissection of the growth plate confirmed an increased expression of Mepe in the hypertrophic chondrocytes (9-fold increase compared to proliferative chondrocytes, P<0.05). ATDC5 cells showed an initial decrease in Mepe expression at day-10 of culture, after which expression increased at day-15. Treatment of ATDC5 cells with pASARM peptide caused an inhibition of mineralisation, determined by alizarin red staining (P<0.01). Treatment with npASARM promoted mineralisation (P<0.01). The growth rate of 15 and 17-day-old embryonic metatarsals and the proliferation of chondrocytes within were not affected by treatment with 20µM pASARM or npASARM peptides. This indicates that the peptides were not toxic and the bones were still viable after 7 days of culture. However, in 17-day-old embryonic metatarsals, the growth of the central diaphyseal mineralisation zone was inhibited in bones treated with 20µM pASARM (P<0.001). This was further examined in 15-day-old embryonic metatarsals in which the central mineralised core only forms after 7 days in culture. Treatment with 20µM pASARM peptides completely prevented mineralisation. Alkaline phosphatase activity was unchanged between the treated and untreated bones, suggesting that the observed inhibition of mineralisation is not a result of decreased enzyme activity. {BR}Our findings indicate that MEPE is expressed by growth plate chondrocytes and that it is likely to have a developmental role in the inhibition of cartilage matrix mineralisation through its cleavage and subsequent phosphorylation of the ASARM peptide. Keywords: Bones, Bone Research Conference: 2011 joint meeting of the Bone Research Society & the British Orthopaedic Research Society, Cambridge, United Kingdom, 27 Jun - 29 Jun, 2011. Presentation Type: Oral Topic: Abstracts Citation: Staines K, Macrae V and Farquharson C (2011). MEPE regulates growth plate mineralisation through its cleavage to the ASARM peptide. Front. Endocrinol. Conference Abstract: 2011 joint meeting of the Bone Research Society & the British Orthopaedic Research Society. doi: 10.3389/conf.fendo.2011.02.00063 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 30 Sep 2011; Published Online: 30 Sep 2011. * Correspondence: Prof. KA Staines, the University of Edinburgh, the Roslin Institute and Royal (Dick) School of Veterinary Studies, United Kingdom, k.staines@brighton.ac.uk Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers KA Staines VE Macrae C Farquharson Google KA Staines VE Macrae C Farquharson Google Scholar KA Staines VE Macrae C Farquharson PubMed KA Staines VE Macrae C Farquharson Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.