Targeted cell-specific deletion of Kir4.1 channels to determine their functions in oligodendrocytes

Lead Research Organisation: University of Portsmouth
Department Name: Sch of Pharmacy & Biomedical Sciences

Abstract

Oligodendrocytes are specialised cells of the brain and spinal cord that form myelin. Myelin is the insulating layer around nerve axons and is essential for rapid conduction of information. As such, the normal functioning of oligodendrocytes is essential for Lifelong Health and Wellbeing. Most oligodendrocytes in the brain are generated around birth and during the first years of development. However, oligodendrocytes continue to be generated in the adult brain and this is important for replacing cells that are lost during normal ageing. Notably, generation of oligodendrocytes in the ageing brain can be inadequate for reparative processes, and this is important in age-related loss of white matter integrity and cognitive function. The mechanisms underlying white matter defects in the ageing brain are unresolved. Now, we have identified a specific cell membrane ion channel called Kir4.1 that is crucial for oligodendrocyte function. We propose to determine the mechanisms by which Kir4.1 regulates the generation of oligodendrocytes in the developing and ageing brain. This work meets the BBSRC Research Priority of Ageing Research (Lifelong Health and Wellbeing), and will make a significant contribution to addressing BBSRC core bioscience and thematic strategic priorities.

Technical Summary

The project is basic research to examine the fundamental importance of the Kir4.1 potassium channel subtype in oligodendrocytes. We have demonstrated that oligodendrocytes strongly express Kir4.1 and that there is severe oligodendrocyte dysfunction following global ablation of Kir4.1 in conventional knock out (KO) mice. However, the precise function of Kir4.1 channels in oligodendrocytes are unknown and it is not clear why they are so critically important for myelin-formation. Our studies highlight the crucial importance of Kir4.1 in oligodendrocytes, but in the global Kir4.1 KO mouse the effects on oligoderocytes represent developmental defects, and it is not possible to distinguish between direct and indirect effects. Determination of the precise functions of Kir4.1 in oligodendrocytes requires targeted disruption of these channels. We will use tamoxifen-inducible CreERT2/loxP gene targeting to create the first inducible KO mice in which the Kir4.1 gene is specifically disrupted in oligodendrocytes and their progenitors. Generation of these mouse strains will enable us to control the temporal and cell-specific expression of Kir4.1, with which we will be able to make major advances in our understanding of the functions of oligodendrocyte Kir4.1 in the developing and ageing brain.

Planned Impact

The main beneficiaries of our research are other scientists, and dissemination of research findings will be through the normal routes of publications, reviews and presentations at meetings. In the longer term, our research is also likely to be of interest to clinicians, patient groups, and the wider public. I will make every effort with the UoP to publicise support we receive from all sources, and to explain our research objectives and potential of the results we gain. When possible, I will make myself available to speak to local and national media on the importance of how grant support drives our much needed fundamental research into glial cells, and how this is relevant to the general public.
It is not anticipated that any directly patentable data will be obtained from this project, but I will continue to work closely with RK&T in the UoP to identify the wider impact and commercialization potential of our research.
It is unlikely that this research will lead to new drug design in the immediate future, but it will hopefully help identify potential targets that will lead to the future development of novel therapies aimed at promoting glial cell function in their critical support of neurons. This would have considerable wider impact. Through the IBBS and Brains Trust, I will use several routes to communicate our findings to the potential user groups and general public. I will present live demonstrations to local schools, and write articles for specialised magazines for patients to whom my research will ultimately be directy relevant.

Publications

10 25 50

 
Description We have: (1) developed a new transgenic mouse strain; (2) discovered novel functions for a number of ion channels and neurotransmitter receptors in promoting neural cell integrity and myelination; and (3) identified key changes in ion channels and neurotransmitters in ageing white matter. These data have been presented at a number of conferences, and to the public in lectures and lab visits. A number of full papers have been published.
Exploitation Route The new functional roles for cellular targets discovered in this project could be taken forward by other academics and the pharmaceutical industry. We have provided evidence that specific ion channels in glia are essential for brain function and integrity.
Sectors Healthcare,Pharmaceuticals and Medical Biotechnology

URL http://www.port.ac.uk/institute-of-biomedical-and-biomolecular-science/cell-biology-and-pharmacology/arthur-butt/
 
Description The findings to date have been used to inform other scientists, members of the public, and strengthen links with industry.
First Year Of Impact 2014
Sector Healthcare,Pharmaceuticals and Medical Biotechnology
Impact Types Societal

 
Description ARUK Pilot Grant - co-PI with Southampton University
Amount £35,000 (GBP)
Funding ID ARUK-PPG2014B-2 
Organisation Alzheimer's Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 12/2014 
End 11/2015
 
Description 2013 Southampton University Neuroinflammation group 
Organisation University of Southampton
Country United Kingdom 
Sector Academic/University 
PI Contribution We are examining human ageing tissue provided by the collaboration
Collaborator Contribution Provision of human tissue and knowledge of human neuropathology
Impact New directions on the changes in glia in the ageing brain resulting in a pilot grant from the ARUK
Start Year 2014
 
Description 2014 Prof Christian Giaume, Paris 
Organisation College of France
Country France 
Sector Academic/University 
PI Contribution We have brought the expertise in oligodendrocyte biology
Collaborator Contribution Prof Christian Giaume has brought the expertise in gap junctions and CNS metabolism. He will also provide the facilities for the planned experiments.
Impact We have been successful in obtaining an Entente Cordiale Fellowship for the PDRA to be seconded to Prof Christian Giaume's lab in Paris to perform a series of key experiments that are not possible in the host institute.
Start Year 2014
 
Description Professor Silvia Lima Costa, Bahia University, Brazil 
Organisation Federal University of Bahia
Country Brazil 
Sector Academic/University 
PI Contribution Experimental design, hosting researchers, writing and publishing papers, hosting and attending bilateral workshops
Collaborator Contribution Funding (Brazil), experimental design, hosting researchers, writing and publishing papers, hosting and attending bilateral workshops
Impact Bilateral meetings and publications.
Start Year 2017
 
Description 2013 Conference (Birmingham, UK) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact IUPS July 21-26, Birmingham, UK.
Expression and function of Kir7.1 in neurons and glia of the CNS.M Papanikolaou, J Hallett, A Lewis, A Butt


Collaborations and increased job prospects for PhD student
Year(s) Of Engagement Activity 2013
 
Description 2013 December Anatomical Society Meeting, Newcastle 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Discussions about the impact of our work and possible future collaborations with a group in Ireland

Academics have visited my lab
Year(s) Of Engagement Activity 2013
 
Description 2013 Laboratory Visit (My labs, Portsmouth) 
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 Public/other audiences
Results and Impact 25 people toured my labs as part of 'Dementia week', which sparked questions on the value of fundamental research to the potential cure of neurological diseases

Feedback indicated new appreciation for the research going on in our university and the value of funds received from research councils and charities
Year(s) Of Engagement Activity 2013
 
Description 2013 Public Lecture (Portsmouth) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact 100 members of the public attended a series of public lectures on 'Inside the World of the Brain', which sparked questions and discussion on the research we do, how it generates new knowledge, and ultimately lead to the development of potential therapies for neurological diseases.

Very positive feedback from members of the public on the value of our fundamental research and monies provided by the research councils
Year(s) Of Engagement Activity 2013
 
Description 2014 June Padue Seminar and visit 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Undergraduate students
Results and Impact 100 undergraduate and postgraduate students attended the talk, together with academics from the University of Padua, which resulted in lively discussions, exchange of ideas, establishment of new collaborations, and discussion of future student exchanges and fellowships.

We have had exchange students from University of Padua working in the PI's lab.
Year(s) Of Engagement Activity 2014
 
Description 2014 March University of Paris 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact New collaboration was established

Exchange of students from France to work in my lab
Year(s) Of Engagement Activity 2014
 
Description 2014 October Dublin Seminar 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact 15 students attended my workshop, which sparked discussions about the significance of glial cells in neuropathology

Students have reported increased interest in glial cells in neuropathology
Year(s) Of Engagement Activity 2014
 
Description 2014 September ISRT visit London 
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 Policymakers/politicians
Results and Impact Meeting with funders of ISRT to promote understanding of spinal research and secure future funding.

Lab visit
Year(s) Of Engagement Activity 2014
 
Description 2014 September IoP Seminar 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Postgraduate students
Results and Impact ~100 students attended my seminars, which sparked questions and discussions

Students have requested further information on glial cell biology and their roles in pathology
Year(s) Of Engagement Activity 2014
 
Description 2014 September SoNG meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact A great deal of interest in the posters presented by the PDRA and PhD students, resulting in job offers and collaborations

New discussions with SAL Scientific, Southampton, to examine potential for grant to NC3Rs
Year(s) Of Engagement Activity 2014
 
Description 2014 September Southampton Dementia conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Considerable interest inn the poster presentations of the PDRA and PhD students, resulting in offers of jobs for them, and potential collaborations

New collaboration with Dr Roxanne Carare, Southampton, on changes in glial cells in human Alzheimer's disease and Bipolar Disorder brain tissue
Year(s) Of Engagement Activity 2014
 
Description 2015 BBB Conference Southampton 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Interaction between basic scientists and clinicians - developed greater understanding of the clinical needs
Year(s) Of Engagement Activity 2015
 
Description 2015 Euroglia Meeting Bilbao 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Presentation sparked a great deal of interest and ideas for future experiments and collaborations

Discussions with other researchers led to considerable interest in utilising our spinal cord slice model
Year(s) Of Engagement Activity 2015
 
Description 2015 FENS Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Presentation sparked many interesting discussions and ideas for future experiments

Discussions with other researchers led to considerable interest in utilising our spinal cord slice model
Year(s) Of Engagement Activity 2015
 
Description 2015 October UPAN Laboratory Tour 
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 Public/other audiences
Results and Impact Talk sparked a great deal of interest and discussions

The members of the public requested further information on our research
Year(s) Of Engagement Activity 2015
 
Description 2015 September ISRT Network Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Poster presentation sparked questions and discussions, leading to potential collaborations and directing future work.

After the presentation, we have established further experiments to be performed.
Year(s) Of Engagement Activity 2015
 
Description International Workshop, Brazil 2013 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Advanced School in Developmental Neurobiology, Rio de Janeiro, Brazil, Nov 25 2013. "Regulation of gliogenesis and neurogenesis in the adult brain", by Arthur M. Butt

Established potential collaborations with the local postgraduate neuroscience programme and identified potential candidates for PhD in spinal research in the UK.
Year(s) Of Engagement Activity 2013
 
Description Living with Dementia, Southampton UK Jan 2016 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Patients, carers and/or patient groups
Results and Impact A mixed audience of PPI, academics, clinicians and funders, with the purpose of promoting understanding and bilateral communication. Considerable interest amongst PPI.
Year(s) Of Engagement Activity 2016