Validating inactivity in the home-cage as a depression-like state indicator in mice

Lead Research Organisation: University of Bristol
Department Name: Faculty of Medical & Veterinary Sciences

Abstract

Non-human animals (hereafter animals) can become strangely inactive in their home pens, being motionless (although awake) and not interacting with their environment. The affective states associated with this waking inactivity are, however, still not well understood. Compellingly, reduced activity and lack of engagement with the environment is one phenomenon supporting clinical depression diagnosis in humans. Moreover, the aetiology of human depression emphasizes aversive life events and chronic stress as common triggers of the disorder, and animals showing waking inactivity have often experienced an array of stressors, such as living in socially and/or spatially restricted environments. We therefore propose that,in animals, specific forms of waking inactivity characterised by being immobile with eyes open represent a marker of depression-like conditions.

This project builds upon our research that has already delineated in mice Mus musculus a specific form of waking inactivity [being awake but motionless within the home cage], triggered by relatively barren lab cages and associated with a key depression-like symptom of the human illness: the tendency to give up when faced with challenge. Our goal is to further test the hypothesis that waking inactivity is a spontaneous depression-like symptom in mice, which would be an easily measured home-cage indicator of animal welfare that could be translated across species.

To achieve this goal, we will adopt a translational 'from humans to animals' approach, using depressed people as a 'model' for inactive mice. As such, potential markers of animal depression-like conditions are expected to co-vary with a range of symptoms in animals similar to those described in human depression. Moreover, for an animal condition to be deemed homologous to a human illness, the processes by which both animal and humans fall into, and are released from, this state must share similarities. If waking inactivity reflects depression-like states in mice, its risk and curative factors should thus be similar to those of the human disease. Our investigations will thus broadly target shared aetiology [is waking inactivity triggered by a combination of risk factors triggering clinical depression in humans?], symptomatic similarities [does waking inactivity co-vary with a range of symptoms similar to those of clinical depression in humans?], and common curative factors [is waking inactivity alleviated by treatments clinically efficient in humans?]. Cognitive symptoms are a frequent part of depression in humans. A quick and easy cognitive measure of depression-like states is however needed in animals, and we will thus first validate such a measure of in mice, and then use this later to assess the symptomatic similarities between waking inactivity in mice and depression in humans.

If confirmed, our results will validate in-cage waking inactivity as a marker of a depression-like condition in mice. This has clear implications for animal welfare, as validated methods of identifying states of mental suffering are first required if we are to detect welfare problems in situ, assess the effectiveness of refinements, and maximise the welfare of lab animals. The ability to detect depression-like states also has crucial implications from a research perspective, since mice in negative affective states may for instance represent improved models of human depression, while potentially invalidating research where stress and depression are not of interest. A new, non-invasive tool to measure affective states will thus be of interest to animal welfare scientists, neuroscientists & psychopharmacologists interested in the assessment of animal affect; biomedical scientists whose research may be impacted by spontaneous occurrence of depression-like states in their subject animals; and laboratory managers & scientists seeking to improve welfare of their research animals.

Technical Summary

This project build upon our previous research & aims to comprehensively investigate the hypothesis that displaying waking inactivity in the home-cage is a depression-like symptom in mice. If confirmed, our results would provide a new, non-invasive tool to assess affective states in the mouse, which has clear implications for animal welfare & refinement purposes. To achieve this goal, we will adopt a translational 'from humans to animals' approach, broadly investigating shared aetiological 'risk' factors (Objective 2), symptomatic similarities (Objectives 1,3) and common curative factors (Objective 4) between in-cage waking inactivity in the mouse and human depression. We will work with C57BL/6J & DBA/2 females (as depression is more prevalent in women), two strains of mice commonly used in research that differ in their tendencies to display both waking inactivity and depression-like features (allowing us to target differences in genetic backgrounds when investigating risk factors, Objective 2). We will use methods from: ethology (e.g. conducting repeated scan-sampling sessions using well-validated methods and ethograms to assess in-cage behaviour and quantify the time spent displaying waking inactivity (Objectives 2-4); welfare science (using behavioural assays to measure affect-related cognitive biases, Objectives 1,3; and more or less enriched home-cages to create comparatively less or more stressful adult life conditions, Objective 2); psychology and psychiatry (applying to mice diagnostic criteria from the Diagnostic Manual of Mental Disorders used in human medicine to investigate symptom similarities, Objective 3); biomedical research (using a maternal separation paradigm to manipulate early life experiences, Objective 2); and pharmacology (using an antidepressant that has been pre-clinically shown to reverse depression-like features in mice and is clinically efficient in depressed people (e.g. venlafaxine) to investigate common curative factors, Objective 4).

Planned Impact

Our goal is to comprehensively test the hypothesis that waking inactivity is a spontaneous depression-like symptom in mice. If confirmed, this will have implications for animal welfare & Refinement purposes. Beneficiaries include:

* Animal welfare scientists, neuroscientists & psychopharmacologists interested in the assessment of animal affect.
Impact will happen via publishing in peer-reviewed journals, attending the leading international conferences in the field & presenting our results at BBSRC UK Animal Welfare Research Network (AWRN) meetings to identify further potential collaborators & future funding opportunities.

* biomedical scientists interest in modelling depression.
Impact will happen via engaging with Bristol psychopharmacologists, clinical epidemiologists & 'Bristol Neuroscience'. Though seminar, meetings & while attending Brain Awareness Week, we will see if our results might be of interest for these communities; if so, we will discuss the best ways to proceed, e.g. presenting the work at tissue & disease focused meetings & via collecting murine samples for pilot translational epidemiological investigations (screening for common biomarker between depressed people & mice displaying waking inactivity using e.g. the Bristol cohort study Avon Longitudinal Study of Parents & Children resource).

* laboratory managers & scientists seeking to further refine the use of their research animals.
Impact will happen via collaborating with Dr Campbell (Bristol, computer vision), to see if there are opportunities to develop a tool based on image analysis to automatically quantify the time mice display waking inactivity. We will collect pilot videos of mice in-cage behaviours, to be analysed by an Engineering student. If the computer vision approach appears feasible, we will collect more videos during year 3 to allow further development. We will apply for funding for subsequent software development and testing (e.g. BBSRC Follow-on, NC3Rs Crack-it funds). Potential industrial partners will be identified through liaison with Home Office inspectors, & publicity using the AWRN, NC3Rs & Understanding Animal Research resources, to identify animal technicians and unit managers, and industry representatives who will be invited to attend a round-table at which we will discuss the work implications and record expression of interest in an automated tool. The longer-term impact goal will be to translate these fundamental results from animal welfare science into practical application to automatically quantify waking inactivity in rodent husbandry facilities. We will also organise a workshop uniting international experts in inactivity displays in several species, to discuss the potential for inactivity to measure affective states; knowledge to be presented at lab animal international conferences, published in specialist journals & translated into a booklet for lab managers.

* We also know from previous engagement activities that there is a *considerable public interest* in animal welfare & in the idea that depression might also happen in non-human animals. Mendl, Fureix & Paul have a strong record of commitment to public engagement activities; we will continue working with the Bristol University Centre for Public Engagement to give talks at events (e.g. Pint of science, Science Café), festivals (e.g. Festival of Nature in coll. with the @-Bristol Science Centre; Brain Awareness Week in coll. with the cross-departmental Bristol Neuroscience group) & at schools (e.g. as part of the STEM ambassador scheme & via the Understanding Animal Research resources for schools). We will also join the Speakeeze platform to increase our visibility as speakers to be invited to be part of events. We will liaise with the Bristol University Public Relations Office to write press releases to accompany new project publications, & post non-technical summaries (public accessible) of our research aims and findings on the AWRN website.

Publications

10 25 50
 
Description We have completed our first programme of work in which we developed a novel methodology for assessing affect-related cognitive biases in sensitivity to positive and negative stimuli in mice. Although mice responded to positive and negative stimuli as predicted in the task, further refinement of the affective manipulation is required to assess the suitability of the task for measuring cognitive-bias. The task highlighted interesting behavioural differences between different strains of mice which we are investigating further in our current work (work to be submitted for publication).

In our ongoing programmes of work, we are investigating the influence of housing enrichment and postnatal environment on the development of inactive and abnormal behaviours in different strains of laboratory mice. Recent collaborations have enabled us to also explore links between these behaviours and various physiological processes including hippocampal neurogenesis and gut microbiota changes.

MAR 2020: We have completed a second programme of work. As predicted if waking inactivity (WI) in mice reflects a depression-like state, we found that the behaviour is triggered in mice by risk factors that cause human depression (i.e. genetic predisposition and exposure to environmental stress), and is alleviated by treatments that are clinically efficient in humans (i.e. antidepressant Venlafaxine and reduced environmental stress). Importantly, pharmacological and environmental treatments appeared equally as effective at reducing WI, which is crucial from a Refinement perspective as increasing home cage enrichments is much more practical in rodent husbandry facilities than applying pharmacological treatment. We currently explore whether combining aversive early life experiences to environmental stress and genetic predispositions triggers WI in mice just as it does with depressed people; and whether WI co-varies with other symptoms that share similarities with those observed in depressed humans (e.g. fatigue, cognitive changes), predicting that WI will provide a quicker and simpler measure that correlates well with these tests.
Exploitation Route The new behavioural test of cognitive bias that we have developed may be translatable to other species, including farm animals. It could also be automated for wider use in a lab animal context. However, further validation is still required.
Waking Inactivity is showing promise as a potential new indicator of negative / depression-like affective state in mice. If so, it could be employed by others as an 'in-cage' welfare indicator. This would be further facilitated by development of automated methods for assessing waking inactivity.
Sectors Agriculture, Food and Drink,Pharmaceuticals and Medical Biotechnology

 
Description University of Plymouth School of Biological and Marine Science PhD Studentship
Amount £54,000 (GBP)
Organisation University of Plymouth 
Sector Academic/University
Country United Kingdom
Start 10/2018 
End 10/2021
 
Title Novel measure of affect-related biases in sensitivity to stimuli in mice 
Description We developed a new behavioural test for assessing affect-related variation in sensitivity to differently valenced distratctors during a non-food motivated task in mice. We are currently submitting this work for publication. 
Type Of Material Technology assay or reagent 
Year Produced 2018 
Provided To Others? No  
Impact No impact as yet. 
 
Description Collaboration with Belgian researchers to develop computer vision methods for data collection 
Organisation Free University of Brussels
Country Belgium 
Sector Academic/University 
PI Contribution We have supplied video recordings and stipulated our requirements for automated data collection for a behavioural test that we are developing.
Collaborator Contribution Our partners have started to develop a bespoke computer vision application to allow us to collect the data that we require from the videos and hence speed up, and standardise, behaviour recording.
Impact This is a multi-disciplinary collaboration. There are no outputs yet - the grant has just commenced.
Start Year 2017
 
Description Collaboration with Bristol computer vision / deep-learning researchers to explore possible use of video and thermal imaging to detect waking inactivity 
Organisation University of Bristol
Country United Kingdom 
Sector Academic/University 
PI Contribution We are collecting, or have collected annotated data on home-cage mouse behaviour hat can be utilised by our colleagues to develop automated methods for detecting waking inactivity
Collaborator Contribution Dr Neill Campbell is exploring the possibility of using video image analysis to detect waking inactivity. Drs John Fennell and Laszlo Talas are developing methods for collecting thermal imaging video to detect waking inactivity.
Impact No outputs as yet. The collaboration is multi-disciplinary involving: behavioural biology; welfare science; computer vision; deep-learning; thermal imaging
Start Year 2019
 
Description Collaboration with Zantiks Ltd (Cambridge) - a new company developing and supplying equipment for behavioural studies. 
Organisation Zantiks Ltd
Country United Kingdom 
Sector Private 
PI Contribution We are developing a collaboration with Zantiks Ltd in which we work alongside them to develop new behavioural assays and associated functionality for their equipment. In particular, we are focusing on assays of mouse and rat behaviour, including the variant of the judgement bias task developed under our NC3Rs grant. We are contributing our expertise in behaviour testing to help Zantiks develop new tests that other researchers may want to use. We are also actively seeking to develop tests that do not require the use of aversive shock stimuli, and are discussing incorporation of an alternative air-puff function into their equipment boxes as a 3Rs benefit of their system. Researchers involved in our current BBSRC grant are assisting with developing and trialling the tests with the new hardware.
Collaborator Contribution Zantiks have supplied us with 4 of their behaviour testing units at a reduced price. They are particularly keen to develop their rat behaviour test system with our input, and also to share other developments with us.
Impact These are early days and, as yet, there are no specific outcomes to report.
Start Year 2018
 
Description Gut microbiota changes as indicators of affect and welfare 
Organisation University of Bristol
Country United Kingdom 
Sector Academic/University 
PI Contribution Collaboration developed with Professor Mick Bailey to investigate associations between environmental enrichment, behaviour. welfare and gut microbiota in different strains of laboratory mice. We will supply faecal samples to Prof Bailey's group for subsequent microbiota analysis and interpretation.
Collaborator Contribution Prof Bailey is highly experienced in gut immunology and has experience and techniques for microbiota analysis.
Impact Early days, so no outputs yet
Start Year 2018
 
Description Hippocampal neurogenesis measures of affect and welfare 
Organisation Newcastle University
Country United Kingdom 
Sector Academic/University 
PI Contribution We have developed a collaboration with Dr Tom Smulders' research group at Newcastle University, who are expert in using hippocampal neurogenesis as an indicator of stress and welfare. We will supply them with brains from mice that were housed in different conditions so that we can assess links between housing treatment, brain measures and our new behavioural measure of affective state in these animals. We will publish collaboratively on any findings that arise.
Collaborator Contribution Dr Smulders' group will carry out assays of hippocampal neurogenesis in the brains that we supply.
Impact No outputs as yet.
Start Year 2018
 
Description Understanding and measuring conscious components of animal affective states 
Organisation University of California, San Diego (UCSD)
Country United States 
Sector Academic/University 
PI Contribution Through meeting at the Lorentz 2017 workshop on Comparative Affective Science (Leiden. Netherlands) at which we presented some of our NC3Rs work, we formed a collaboration with Profs Piotr Winkielman and Marco Tamietto and Dr Shlomi Sher to review and develop thinking on challenges and potential ways forward in assessing conscious affective states in animals. We are sharing our ideas and writing a substantial review on the topic.
Collaborator Contribution Discussions and joint writing of a substantial review article on the topic.
Impact Paper on conscious component of animal affect. Paul, E.S., Sher, S., Tamietto, M., Winkielman, P. & Mendl, M.T. (2020). Towards a comparative science of emotion: affect and consciousness in humans and animals. Neuroscience and Biobehavioral Reviews 108, 749-770
Start Year 2017
 
Description Understanding links between inactivity and affective state 
Organisation University of Guelph
Country Canada 
Sector Academic/University 
PI Contribution Sharing plans, methods, and findings from our ongoing research investigating waking inactivity as an indicator or affective states
Collaborator Contribution Prof Georgia Mason and her team (University of Guelph) are sharing plans, methods, and findings from ongoing research investigating waking inactivity as an indicator or affective states
Impact No outcomes as yet.
Start Year 2018
 
Description Animal welfare science stall at 'We The Curious', Bristol 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact We ran a stall, based at We The Curious (Bristol) with the aim of engaging school children with animal welfare science.
Year(s) Of Engagement Activity 2018
 
Description Emily Finnegan gave a talk to the 3rd University of Bristol 3Rs Symposium, Bristol, UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Talk given at a 3Rs meeting attended by researchers, lab technicians and postgraduate students.
Year(s) Of Engagement Activity 2019
URL https://www.bristol.ac.uk/news/2019/august/3rs-symposium.html
 
Description Interview for German TV production company 'nonfiction society' on animal emotion and the ethics of animal treatment 
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 German TV crew came to Bristol to interview me about animal emotions, how one might go about studying them and assessing them scientifically, and what the implications of understanding animal affective states may have for our treatment of animals in many contexts (e.g. laboratories, farming etc.)
Year(s) Of Engagement Activity 2019
URL https://www.nonfictionsociety.de/
 
Description Invited speaker: Animal Welfare Science Centre Seminar, University of Melbourne, Australia 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Invited talk to postgraduate students and other researchers at the University of Melbourne Animal Welfare Science Centre
Year(s) Of Engagement Activity 2019
 
Description Invited talk at Melbourne Zoological Gardens, Melbourne, Australia. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact invited talk to zoo keepers and other zoo staff, plus postgraduate students at Melbourne Zoo, Australia
Year(s) Of Engagement Activity 2019
 
Description Invited talk by E. Paul to Further Education Students at Plumpton College, East Sussex 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Undergraduate students
Results and Impact Talk given to FE students on "Measuring Emotion in Animals"
Year(s) Of Engagement Activity 2019
 
Description Keynote Speaker at Tecniplast Animal Welfare Symposium, Australian and New Zealand Laboratory Animal Association Annual Meeting, Australian National University, Canberra, Australia. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Keynote speaker at ANZLAA Annual Meeting attended by researchers, laboratory technicians, industry representatives, postgraduate students, policymakers
Year(s) Of Engagement Activity 2019
URL https://www.anzlaa.org/resources/Documents/2019%20AWS%20Brochure%20Final.pdf
 
Description Keynote Speaker at the Midlands 3Rs Symposium, University of Leicester 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Keynote speaker to an audience of researchers, laboratory technicians, postgraduates and funders.
Year(s) Of Engagement Activity 2019
URL https://twitter.com/hashtag/Midlands3Rs?src=hashtag_click
 
Description M. Mendl interviewed for German non-fiction society TV company (www.nonfictionsociety.de) for documentary on animal emotion and the ethics of animal treatment 
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 German non-fiction society TV company (www.nonfictionsociety.de) made a documentary on animal emotion and the ethics of animal treatment in which M. Mendl appeared as an expert in the area referring to relevant experimental studies. Documentary was seen by about 1 million people and is available online at: https://www.3sat.de/wissen/wissenschaftsdoku/rechte-der-tiere-100.html The film-makers said that it was well received but I don't have figures on any impact on the audience.
Year(s) Of Engagement Activity 2019
URL https://www.3sat.de/wissen/wissenschaftsdoku/rechte-der-tiere-100.html
 
Description M. Mendl interviewed for University of Melbourne, Australia 'Encounters with Experts' podcast (15 May 2019). 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Interview detailed approaches to assessing animal affect and welfare and mentioned relevant research work. Interviewer, Andi Horvath, said: "The Professor was a fabulous speaker. One of the best interviewees I've ever had. I so enjoyed his thoughtful approach to research very enlightening!". Editor Chris Hatzis said: "The interview turned out very well. A very illuminating chat". Communication and Content Coordinator Stuart Winthrope said: "That was a great listen, thank you".
Year(s) Of Engagement Activity 2019
URL https://pursuit.unimelb.edu.au/podcasts/how-can-we-tell-if-an-animal-is-depressed
 
Description Opening keynote: Introduction to animal emotion: concepts, correlates and consciousness. The Fundamentals of Animal Emotion Workshop, Wageningen University and Research, Netherlands. 
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 Opening keynote speaker and tutor at workshop on the assessment of animal affective states for PhD students from all over Europe.
Year(s) Of Engagement Activity 2019
URL https://www.wur.nl/upload_mm/c/d/9/6fdfd010-c2e2-47d4-a605-ae15b0f5ec93_Programme%20PhD%20course%20T...
 
Description Organiser of the Association for the Study of Animal Behaviour 2018 Winter Meeting, London 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact We organised the annual ASAB Winter Meeting in London on the topic of 'Behavioural Biology in Animal Welfare Science'. Over 200 delegates attended across the two days and talks, including 4 invited plenaries, were given by researchers from all over Europe with a few attending from North America. The audience were mainly academics, postgraduate research students and some undergraduates working in the areas of animal behaviour and animal welfare science. The meeting included poster sessions and resulted in discussions and increased interest in animal welfare research by those whose previous focus was mainstream animal behaviour. New collaborations also resulted from the meeting.
Year(s) Of Engagement Activity 2019
 
Description Stall at 2020 University of Plymouth Science and Technology Showcase 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Stall developed and manned with the aim of engaging school children with animal welfare science and how to measure emotions in animals.
Year(s) Of Engagement Activity 2019
URL https://www.plymouth.ac.uk/study/outreach/science-technology-showcase
 
Description Talk by Emily Finnegan to University of Bristol Technicians Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Talk given to conference attended by University of Bristol animal technicians.
Year(s) Of Engagement Activity 2019
 
Description The Charles River Lecture, American Association for Laboratory Animal Science Annual Meeting, Baltimore, Maryland, USA 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited to give the annual Charles River Lecture at the AALAS meeting in Baltimore, USA in 2018. Meeting attended by researchers, laboratory technicians, postgraduate students, industry representatives and salespeople, policy makers, third party sector organisations.
Year(s) Of Engagement Activity 2018
URL https://www.nc3rs.org.uk/events/69th-aalas-national-meeting
 
Description Theory and practical applications in the assessment of laboratory animal welfare. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Talk given to the University of Guelph, Canada, IACUC grouping and animal technicians on the assessment of animal affect and welfare and the theory behind it.
Year(s) Of Engagement Activity 2018
 
Description University of Bristol 3Rs Symposium 2018 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact We attended the 2nd University of Bristol 3Rs workshop and presented posters of our ongoing work in laboratory rodents and other species.
Year(s) Of Engagement Activity 2018