Join Lior Bikovski and Shivang Parikh from Tel-Aviv University for a presentation on longitudinal behavioral studies and how to optimize the use of home cage monitoring (HCM) systems for behavioral research. Multiple home cage monitoring systems have been developed during the last two decades with the aim of increasing the reproducibility of test results and improving behavioral assessment in pre-clinical research. In this webinar, Lior Bikovski and Shivang Parikh will address specific advantages and limitations of today’s home cage monitoring (HCM) technology used in behavioral research. Specifically, they will discuss the use of HCM systems and compare them with other standard tools in the field of behavioral research. They will explain why it can be difficult to see differences between study groups using point sampling methods and why longitudinal tools can be helpful in characterizing behaviors. They will also review the calibration of a new system, preliminary results, and the benefits of using the home cage as the test chamber, rather than moving animals to a separate test chamber for observation experiments. Finally, Shivang will address his current work involving HCM and the effects of ultraviolet light with a focus on behavioral assessments and cancer. Key Topics Include: - What is a home cage monitoring (HCM) system - Two main categories of HCM systems - Benefits of using HCM for behavioral research - Examples of data that can be acquired using HCM

1. Lessons From the Core:
Longitudinal Assessment
vs. Sampling of Behavior
Shivang Parikh
Ph.D. Student
Tel-Aviv University, Israel
Advisor: Professor Carmit Levy
Dr. Lior Bikovski
The Myers Neuro-Behavioral
Core Facility
Tel-Aviv University, Israel

2. Lessons From the Core:
Longitudinal Assessment
vs. Sampling of Behavior
Experts discuss behavioral phenotyping
with a focus on the benefits and
applications of home-cage monitoring.

3. Dr. Lior Bikovski
The Myers Neuro-Behavioral
Core Facility
Tel-Aviv University, Israel
Lessons From the Core –
Longitudinal Assessment vs.
Sampling of Behavior
Copyright 2020 L. Bikovski and InsideScientific. All Rights Reserved.

4. TAU Neurobehavioral Core Facility
The MNBCF was Established in 2015 and it is one of 2 facilities in Israel
http://sicf.tau.ac.il/bu/

5. In Vivo In VitroPrenatal
Imaging
Core
Neuro-
Behavioral Core
Histology
Core
Irradiation
Core
Single cell
Genomics
Core
Virus
production
core
General Lab
Instrumentation
Flow
Cytometry
Core
Flow
Cytometry
Core
Center for
Transgenic
Mice
Imagine
Core
Single cell
genomics
Core

6. What is a “standard” sampling tool?
Standard Phenotyping Process
Sampling of behavior in the lab
Standard – as these are the tools we used for the last 50 years
Sampling – as every mouse is sampled for a limited time…

7. Another property of standard tools is STRESS
In some mouse models, abnormal behavior tends to be subtle, and in some cases mice could
compensate to hide signs of poor health from potential predators or under specific elicit test conditions.
Low
High
High
Performancelevel
Amount of stress
Optimum
level
Simple task
Open field test – difference
between 2 strains of mice

8. Compensation in exploration/motor behavior
Assessments of Motorexploration behavior: significant differences seen at the
phenotypers (longitudinal) are absent in the open field (sample).

9. Another property of standard tools is STRESS
The advantage of the standard tools is the possibility of using maximum stress when needed
Low
High
High
Performancelevel
Amount of stress
Optimum
level
Aversive task
1. Fear-conditioning
2. Water-maze
3. Treadmill

10. Longitudinal Assessment Tools
Access to testing
unit
Monitor mice in
cage
PubMed search for “home-cage” and “mice” show
increased use of home cage monitoring

11. Access to testing
unit
Monitor mice in
cage
Total motor activity (running wheels or distance traveled) was
the same but activity patterns were different
https://link.springer.com/article/10.1007/s12031-019-01422-9
PubMed search for “home-cage” and “mice” show
increased use of home cage monitoring
Longitudinal Assessment Tools

12. PubMed search for “home-cage” and “mice” show
increased use of home cage monitoring
Access to testing
unit
Monitor mice in
cage
https://www.frontiersin.org/articles/10.3389/fnbeh.2020.575434/full
Longitudinal Assessment Tools

13. PubMed search for “home-cage” and “mice” show
increased use of home cage monitoring
Access to testing
unit
Monitor mice in
cage
https://www.frontiersin.org/articles/10.3389/fnbeh.2020.575434/full
Longitudinal Assessment Tools

14. Access to testing
unit
Monitor mice in
cage
Low stress – assessment in home cage
with minimum human contact
Automated*
PubMed search for “home-cage” and “mice” show
increased use of home cage monitoring
Longitudinal Assessment Tools

15. Low stress – assessment in home cage
with minimum human contact
Automated*
Spoiler alert: automated is not “Fire And Forget”
PubMed search for “home-cage” and “mice” show
increased use of home cage monitoring
Access to testing
unit
Monitor mice in
cage
Longitudinal Assessment Tools

16. Longitudinal – continuous assessment
(in day differences and between days)PubMed search for “home-cage” and “mice” show
increased use of home cage monitoring
Access to testing
unit
Monitor mice in
cage
Low stress – assessment in home cage
with minimum human contact
Automated*
Longitudinal Assessment Tools

17. Stories from the Core – motor
assessment in mTBI mice
Example for compensation effects in mice
Chagi Pick
Namdar I et al. Motor Effects of Minimal Traumatic Brain Injury in Mice. Journal of Molecular Neuroscience, 1-13 (2020).

18. Stories from the Core – motor assessment in mTBI mice
RotaRod - Balance and learning
Open field - General motor ability
Treadmill - Max motor ability
Grip Strength - Muscles function
• mTBI mice showed no motor
deficiencies (unlike the Human
condition).

19. Compensation techniques used by
mTBI mice
CatWalk gait analysis
https://www.noldus.com/catwalk-xt

20. Stories from the Core – motor assessment in mTBI mice
mTBI mouse
Control mouse
(A) BOS in controls - (B) BOS in TBI mice
A
B

21. Stories from the Core – motor assessment in mTBI mice
RotaRod - Balance and learning
Open field - General motor ability
Treadmill - Max motor ability
Grip Strength - Muscles function
• mTBI mice showed no motor
deficiencies (unlike the Human
condition).
• mTBI mice showed no anxiety
deficiencies (unlike the Human
condition).
0
20
40
60
80
100
120
140
Control mTBI
Timeinopenarms(sec)
days
EPM

22. So we turned to the home-cage
system

23. Phenotyper cage – motor and well-being
assessment protocol
https://www.noldus.com/phenotyper

24. Phenotyper cage – motor and well-being
assessment protocol
Time spent frequency
• In shelter (safe zone)
• On shelter (risky zone)
• Food water intake
Motor assessment
• Distance traveled
• Activity
• Running wheel
Specific measured behaviors (e.g. Grooming, jumping)

25. Stories from the Core – tracking motor
behavior in mTBI mice
Dark/active PhaseLight/rest Phase
Motor/exploration
information –
Distance moved in
cm
No motor
exploration
differences
Running wheel results

26. Distance moved as indication for circadian
rhythm integrity

27. Stories from the Core – tracking motor
behavior in mTBI mice
Dark/active PhaseLight/rest Phase
Motorexploration
information –
Distance moved in
cm
Psychiatric
assessment – Time
spent in shelter in
seconds
No motor
exploration
differences
mTBI mice spend
more time in shelter
(anxietydepression
motivation)
Floor effect *

28. Assessment of cognition in
the home cage system
Cognition wall

29. Cognition wall – cognitive assessment
false
True
false
Picture adopted from - https://www.sylics.com/bioinformatics/news/

30. Cognition wall – cognitive assessment
false
True
false

31. Preliminary results in cognitive enhancer drug
Tania Kudinov
Drug
Drug
Control
Control

32. Touchscreen Chamber, home cage system (TCHC)
Picture adopted from Rivalan et al, 2017 - https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0169476
PhenoSys GmbH
https://www.phenosys.com/products/touchscreen-chamber/

33. Touchscreen Chamber, home cage system (TCHC)
PhenoSys GmbH
Home cage
3 sorters in a
tunnel (RFID)
The
Touchscreen
chamber

34. Touchscreen Chamber, home cage system (TCHC)
Liron Tseitlin
PhenoSys GmbH
Home cage
3 sorters in a
tunnel (RFID)
The
Touchscreen
chamber
• Around 5-6 mice (strain dependent)
• Detection - RFID chips (only in sorter)
• 14 days of learning the task + 1-3 days of test.
• Learning = 5 different steps
• Motivation is food pellets

35. TCHC protocol
• General Touchscreen pre-training (GTPT):
• GTPT1 Habituation
• GTPT2 Initial touch
• GTPT3 Must touch
• GTPT4 Must initiate
• GTPT5 Punish incorrect
• Test – delay non match to sample

36. The test protocol
Pre session
Exposure
to sample
Delay
Non match

37. The test protocol
Pre session
Exposure
to sample
Delay
Non match

38. Data and results
• Process of learning – how many trials for criteria?
• Result of learning (Test) - how many trials for criteria?
• Why trials and not time?
Traininglearning process Test phase
N = 3-4

39. One important limitation of
attached testing unit
(And of group housing in home cage systems)

40. Limitations – learning in social groups
8:30-12:30 12:30-16:30 16:30-20:30 20:30-00:30 00:30-4:30 4:30-8:30
Control 1 1 2 0 2 0 0
Control 2 0 0 1 1 3 0
Control 3 2 1 1 1 0 0
mTBI 5 0 1 2 2 0 0
mTBI 6 3 0 0 1 1 0
mTBI 7 2 3 0 0 0 0
mTBI 8 1 1 1 0 1 1
0
1
2
3
4
5
Numberofentrances
Time
Learning schedule per day
Control 1 Control 2 Control 3 mTBI 5 mTBI 6 mTBI 7 mTBI 8
Dominant mouse
Submissive
Access 4 hours after rest
phase started

41. Conclusions
• Standard tests are very important (elicit behaviors, gold standard).
• Home-cage assessment is a longitudinal assessment for a condition
(hours – days – weeks) which allows us to get a more holistic view.
SpiroSmart - a research app for
lung function (Univ. of Washington)
Standard lab - Exercise stress
test
+

42. Shivang Parikh
Ph.D. Student
Tel-Aviv University, Israel
Advisor: Professor Carmit Levy
A Story From the Core:
Deciphering UVB Exposure
Effect On Behavioral Appetite
Changes
Copyright 2020 S. Parikh and InsideScientific. All Rights Reserved.

43. Being a biologist, why studying BEHAVIOUR is important?
The capacity and potential to express and respond to external or internal stimuli.
Factors affecting :
Mental
Physical
Social
Factors affecting :
Environmental cues
Society
Factors affecting :
Anatomical
Physiological
Biochemical
Molecular

44. ARPANSA, Australian Government (2015)
UV induced effects Type of effect
DNA damage
- DNA lesions and repair mechanism (Bioessays, 1996).
- UV-induced DNA lesions can lead to mutations in the DNA
sequences (PNAS, 1992).
Hair follicles
- Modifies hair growth and cycle, promotes cell death, and induces
complex regulatory events (Cell, 2007).
- Still unclear in vivo conditions.
Skin Pigmentation
-Regulation of human skin pigmentation and responses to ultraviolet
radiation (Pigment Cell Res, 2007).
Fatigue
-UV damage to skin stimulates production of 2- endorphin, which is
addictive (Cell, 2014).
UV EXPOSURE & IT’S EFFECTS

45. epidermisdermis
Oren & Bartek, Cell (2007).
Fell et al., Cell (2014).
WHAT HAPPENS AFTER UV HITS THE SKIN ?

46. How can we measure the appetite related behavioural changes?
Are these changes prevalent in both males and females?

47. WEEK 4WEEK 3WEEK 2WEEK 1HabituationMice Arrived
Control UV
Mock
UV
UVB induce pigmentation
0
50
100
150
0 1 2 3 4
Meanmelaninlevels
Time after first UV (Weeks)
Mock
UV
***
*** *** ***
Control
UV
0
50
100
150
0 1 2 3 4
Meanmelaninlevels
Time after first UV (Weeks)
Mock
UV
***
***
*** ***
Control
UV
Parikh et al., in Revision

48. WEEK 8WEEK 7WEEK 6
Chronic UV exposure
~ 5 WEEKS
HabituationMice Arrived
ANIMAL BEHAVIOURAL TESTS
Experimental design for animal behavioural studies

49. Control UV
Control UV
0
1
2
3
4
5
6
Eaten Attempted
Mock
UV**
Control
UV
Number of food pellets
0
1
2
3
4
5
6
Eaten Attempted
Mock
UV
Control
UV
Number of food pellets
Staircase test for appetite related-animal behavioural studies
Parikh et al., in Revision

50. Open-field test for appetite related-animal behavioural studies
0
0.1
0.2
0.3
0.4
0.5
0.6
Activity (% pixel
change)
0
2000
4000
6000
Total distance (cm)
**
0
100
200
300
400
500
Total time spent in
center (sec)
***
0
2000
4000
6000
8000
Total distance (cm)
0
5
10
15
20
25
Pellets eaten
(number)
***
0
10
20
30
40
Number of center
visits (Frequency)
0
2
4
6
8
10
Velocity (cm/sec)
**
0
0.1
0.2
0.3
0.4
0.5
0.6
Activity (% pixel
change)
*
0
10
20
30
40
Number of center
visits (Frequency)
0
2
4
6
8
10
Velocity (cm/sec)
Control
UV
Center
Center
Control
UV
Center
Center
0
5
10
15
Pellets eaten
(number)
Parikh et al., in Revision

51. 0
500
1000
1500
2000
2500
3000
3500
4000
4500
1 2 3 4
Lickometer(N)
Time after first UV (Weeks)
Control Active
Control Rest
UV Active
UV Rest
0
100
200
300
400
500
600
700
800
1 2 3 4
Beambreaks(N)
Time after first UV (Weeks)
Control Active
Control Rest
UV Active
UV Rest
0
100
200
300
400
500
600
700
800
1 2 3 4
Beambreaks(N)
Time after first UV (Weeks)
Control Active
Control Rest
UV Active
UV Rest
**
*
*
***
**
0
500
1000
1500
2000
2500
3000
3500
4000
4500
1 2 3 4
Lickometerr(N)
Time after first UV (Weeks)
Control Active
Control Rest
UV Active
UV Rest
*
WEEK 4WEEK 3WEEK 2WEEK 1HabituationMice Arrived
Control UV
UVB induce appetite related changes in mice
Parikh et al., in Revision

52. Bridging the link between Behaviour studies and Cancer Biology
Science, 2013
Nature Clinical Practice Oncology, 2005
Cell, 2014

53. Tel-Aviv University
Professor Carmit Levy
Dr. Lior Bikovski
Roma Parikh
Professor Shamgar Ben-Eliyahu
Professor Segev Barak
Acknowledgements
Bar-Ilan University
Professor Aron Weller
Dalya Gringer Fund

54. Thank you for participating!
Shivang Parikh
Ph.D. Student
Tel-Aviv University, Israel
Advisor: Professor Carmit Levy
Dr. Lior Bikovski
The Myers Neuro-Behavioral
Core Facility
Tel-Aviv University, Israel
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