Join Drs. Jason Rogers and De Wet Wolmarans for a deep dive into two research protocols for tracking, quantifying, and analyzing behaviors in rodent models. Don’t Look Down: Stress Behavior in Long-Evans Rats On A Novel Elevated Open Field Jason Rogers, PhD The open field and elevated plus maze are standard tests to study anxiety or stress-related behaviors in rodents. It is often recommended to use both tasks as a battery; however, sometimes animals display anxiety-like behaviors in one test but do not in another. To combat these issues, the current experiment used a hybrid, elevated open field, to assess the anxiety behaviors in rats who underwent chronic unpredictable stress (CUS). The data show that after CUS, animals spent more time in the center of the apparatus and less time at the edge. In addition, they had a longer latency to explore the edge and had fewer head dips. Finally, the CUS animals maintained more distance from the edge but had no difference from controls in overall distance traveled or speed. Taken together, these data show how automated systems like EthoVision XT can be used to develop and define novel techniques for the study of rodent behavior. Individuality in Naturalistic Deer Mouse Behaviors – Perspectives on Psychobiology De Wet Wolmarans, PhD Studies of rodent behavior contribute to a better understanding of human psychobiological processes. Behavior that deviates from what is regarded as ‘normal’ can generally be induced or studied under spontaneous circumstances. With respect to the latter research framework, near accurate separation between what is ‘normal’ and ‘abnormal’ is vital. In this presentation, closer attention is afforded to some biobehavioral differences that are observed between animals of the same laboratory-housed wildtype mouse species, the deer mouse (Peromyscus maniculatus bairdii). Further, some focus is afforded to the manner in which such behaviors are observed, quantified, and then applied against a translational background of conditions characterized by persistent behaviors, like obsessive-compulsive disorder. The presentation concludes by highlighting a need for the individualized analysis of animal behaviors in the study of naturalistic behavioral expressions as model systems that are aimed at improving our understanding of human conditions. Key Topics Include: - Using the Elevated Open Field to assess stress/anxiety - Effects of chronic unpredictable stress on anxiety-like behaviors in rats - Characteristics of naturalistic behaviors commonly observed in laboratory-housed mice - Importance of accurate analysis of naturalistic animal behaviors, although subjective, to inform our understanding of spontaneous psychobiological trajectories

1. Welcome!
Innovative Approaches to
Tracking and Quantifying
Behavior in Rodents
Jason Rogers, PhD
Senior Research Scientist
Noldus Information Technology
De Wet Wolmarans, PhD
Associate Professor & Head of Research
Translational Neuroscience & Neurotherapeutics
North-West University, South Africa

2. Copyright 2022. All Rights Reserved. Contact Presenter for Permission
Senior Research Scientist
Noldus Information Technology
Don’t Look Down:
Stress Behavior in Long-Evans
Rats On A Novel Elevated
Open Field
Jason Rogers, PhD

3. Don’t Look Down:
Stress Behavior in Long-Evans Rats On
A Novel Elevated Open Field
Jason Rogers, Ph.D.
Thursday, 12 May 2022

4. Who am I
Doctorate in the neurobiology of learning &
memory from the University of Utah. Post-doc
in neuropsychopharmacology at MUSC. Joined
Noldus in 2008 at Lead Trainer. Has been in
services, sales, consulting, and marketing.
Don’t Look Down | 4
 H-Index of 11 (last published in 2008)
 Been to 49 US states
 Has run three marathons (4th scheduled for 2023)
 Amateur woodworker & musician
Jason Rogers, Ph.D.
Random facts
Introduction

5. 1
2
3
4
5
6
Overview
Don’t Look Down | 5
Elevated Open Field
Experiment Rationale & Setup
EthoVision XT: Dynamic Subtraction
EthoVision XT: Deep Learning
Results
Q&A
Today’s Webinar

6. Experimental Approach
 Current project was the behavioral component of a larger Master’s project
 Hypothesis: Do stressed animals self-administer more cocaine (spoiler alert: yes)
 Procedure:
› Long Evans rats (n = 16)
› Jugular catheter implantation
› Chronic unpredictable stress (5 days)
› Cocaine self-administration (14 days)
› Extinction (10 days)
› Reinstatement (3 days)
› Blood draws for corticosterone levels
› Elevated open field testing
Don’t Look Down | 6
Methodology
Jian Xiang Ding, Patrick T. Rudak, Wataru Inoue, S.M. Mansour Haeryfar,
Physical restraint mouse models to assess immune responses under stress with
or without habituation, STAR Protocols, Volume 2, Issue 4, 2021, 100838,
ISSN 2666-1667, https://doi.org/10.1016/j.xpro.2021.100838.

7. Elevated Open Field
 Round open field (122 cm) painted grey
 Elevated 1.5m above the ground
 Anxiogenic environment that forces the rats to
explore while trying to avoid
 No thigmotaxis possible
 Outcome measures:
› Time in center
› Time along the edge
› Latency to explore the edge
› Overall distance traveled and speed
 Animals tracked via EthoVision XT
| 7
Should I Stay Or Should I Go
Don’t Look Down

8. EthoVision XT
 Videos were created and first analyzed in EthoVision XT 5
 Videos were re-analyzed in EthoVision XT 16
 Dynamic subtraction: EthoVision XT compares each sampled image with a reference image, with the
important difference that the reference image is updated regularly. This compensates for any temporal
changes in the background.
| 8
Tracking Parameters
Don’t Look Down

9. EthoVision XT 16 With Deep Learning
EthoVision XT uses a trained network, that is, the network has learned to extract features from a number
of video images of rodents, where the nose and the tail-base were previously annotated. During tracking,
the network analyzes a portion of the image that includes the detected subject. It creates a map of
probability of occurrence for both the nose- and the tail-base points, and finally makes an estimate of the
position of the nose point and the tail-base point, based on the highest probability.
| 9
Tracking Parameters
Don’t Look Down

10. Stressed Animals Don’t Go To the Edge As Much
| 10
Results
Don’t Look Down

11. Stressed Animals Stay In The Center Longer
| 11
Results
Don’t Look Down

12. No Difference In Distance Traveled or Velocity
| 12
Results
Don’t Look Down

13. Deep Learning Is More Accurate & Robust
| 13
Results
Deep Learning
Dynamic Subtraction
Don’t Look Down

14. Conclusions
 Chronically-stressed animals have a higher
latency to explore the edge, spend less
time at the edge, maintain a higher
average distance from the edge, and
spend more time in the center in a novel
elevated open field
 In addition, stressed animals have fewer
head dips and spend less time looking
over the edge
 EthoVision XT with Deep Learning is more
robust and more accurate for nose
detection
| 14
What Does It All Mean
Don’t Look Down

15. We’d love to hear from you
E-mail address
Jason@Noldus.com
Website
www.noldus.com
Find us on social media
| 15
Get In Touch
Don’t Look Down

16. Copyright 2022. All Rights Reserved. Contact Presenter for Permission
Associate Professor & Head of Research
Translational Neuroscience & Neurotherapeutics
North-West University, South Africa
Individuality in Naturalistic
Deer Mouse Behaviors –
Perspectives on
Psychobiology
De Wet Wolmarans, PhD

17. NATURAL PERSISTENCE IN DEER
MICE –
THE IMPORTANCE OF
INDIVIDUALITY
De Wet Wolmarans, Ph.D.
Associate Professor, Department of Pharmacology
North-West University, Potchefstroom
Email: dewet.wolmarans@nwu.ac.za

18. BACKGROUND AND HISTORY
• Peromyscus maniculatus bairdii – deer mouse, origin North-America
• Original stock – University of South Carolina, obtained 2005/6, replaced 2018
Initial validation
Re-evaluation
of validity
Phenotypic
characterisation
Neurobiological
study
Cognitive study
2007-2010 2011-2013 2014-2019 2020-present

19. BEING SUBJECTIVELY
OBJECTIVE

20. IT’S A MATTER OF MINORITIES…

21. IT’S A MATTER OF MINORITIES…

22. IT’S A MATTER OF MINORITIES…

23. IT’S A MATTER OF MINORITIES…

26. A MATTER OF MINORITIES…

29. MINORITIES REVEAL MAJORITIES OF
FINDINGS
Neurobiological and cognitive
perspectives

30. THE MAGIC OF INDIVIDUALITY

31. THE MAGIC OF INDIVIDUALITY

32. THE MAGIC OF INDIVIDUALITY

33. THE MAGIC OF INDIVIDUALITY

34. THE MAGIC OF INDIVIDUALITY

35. THE
MAGIC
OF
INDIVIDUALITY

36. THE MAGIC OF INDIVIDUALITY

37. THE MAGIC OF INDIVIDUALITY

38. THE MAGIC OF INDIVIDUALITY

39. SUMMARY
Individuality in animal behaviours is key
Though subjective, clear separation between phenotypes
are vital
Accurate data are sometimes missed in the bigger scheme

40. THANK YOU!
De Wet Wolmarans
Centre of Excellence for Pharmaceutical Sciences
Subprogramme: Translational Neuroscience and
Neurotherapeutics
Department of Pharmacology
North-West University
+27 (0) 18 299 2230
dewet.wolmarans@nwu.ac.za

41. Thank you for participating!
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