LSID: http://syli.cz/urn:lsid:public.sylics.com:experiment:5EA7-GB53-59D8
| Treatment | Amount | Administration route | Administration time |
Treatment info:
Cohort(s)
Cohort 1, designated Batch 1 on the website
Born between 17-20 November 2011, from HZ x HZ breedings in the MouseHouse
Testing protocol
Standard test battery.
Spontaneous behavior in the PhenoTyper
Data files of 9 WT and 14 HOM data files passed all Quality Control criteria.
With respect to spontaneous behaviors in the first three days in the PhenoTyper, 6 groups of behavioral parameters are defined as described below. The first two groups describe specific behavioral elements related to kinematics of mice (description of movement characteristics, group1) and sheltering behavior (group 2). These 2 groups of behavioral parameters were analyzed with respect to temporal aspects, in particular over 4 different time scales, i.e., habituation effects across multiple days (group 3), effects of DarkLight phase across 24h (group 4), differences in the pattern of behavior in the few hours before and after phase shifts (group5), and differences in activity bout properties on the sub-minute time scale (group 6).
Of each of the 6 groups, one or a few key parameters are described (20 parameters in total). These 20 parameters are selected on the bases of their high correlation with other parameters in their group, indicating that these selected parameters are well-representing the other parameters of the respective group (Loos et. at., in prep.). The total set of spontaneous behavior parameters is 115 parameters. By statistically evaluating the condensed set of 20 parameters rather than the total set of 115 parameters, the multiple testing issue is reduced.
1. Key kinematic parameters: Mice make short movements, such as turning or rearing against the wall, as well as long movements when mice travel from one location in the cage to the next. These two types of movement, i.e. short and long, can be distinguished by plotting the frequency distribution of all move segment distances over three days in the cage, yielded a bimodal distribution. An animal-centered threshold that was defined by the intersection of the two Gaussian distance curves obtained after Gaussian mixture model fitting of data of each individual mouse. Most arrest durations are relatively short, and occur in between move segments when mice reorient themselves. In addition, mice infrequently made long arrests when they were consuming food or water, or when they were resting outside their shelter. An animal-centered threshold was set distinguishing the 90% shortest arrests (short) from the 10% longest arrests (long), separating brief from long arrests.
Long movement max. velocity (cm/s): Average velocity of the 95th percentile fastest long movement segments
Ttc39 KO mice show a higher maximum velocity during long movements
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=54&e=9&batch=0&t=&texp=&list=batches&tdose=
Long arrest threshold (s): Cut-off value to separate short and long arrests
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=55&e=9&batch=0&t=&texp=&list=batches&tdose=
Mean long arrest duration - light (s): Mean duration per long arrest during the light phase
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=123&e=9&batch=0&t=&texp=&list=batches&tdose=
Long movement threshold (cm): Cut-off value to separate short and long movements
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=140&e=9&batch=0&t=&texp=&list=batches&tdose=
2. Key sheltering parameters: Mice frequently visit the shelter for a few seconds (i.e., passing through) during bouts of activity. In contrast, long shelter visits during which mice appeared to be resting or sleeping, are in the range of hours. Although long shelter visits were infrequent, they appeared as a separate class of events when the frequency distribution of shelter visit durations of the three days in the cage was plotted. Long shelter visits were readily identified by Gaussian mixture model fitting of shelter visit data of individual mice. The 90th percentile of the first fitted Gaussian was used as upper threshold to consistently distinguish short shelter visits.
Short shelter visit threshold (s): Cut-off value to separate short and intermediate shelter visits
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=113&e=10&batch=0&t=&texp=&list=batches&tdose=
Long shelter visit threshold (s): Cut-off value to separate intermediate and long shelter visits
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=115&e=10&batch=0&t=&texp=&list=batches&tdose=
Long shelter visit fraction of total visits (fraction): The fraction of shelter visits with duration longer than long shelter visit threshold
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=114&e=10&batch=0&t=&texp=&list=batches&tdose=
Long shelter visit duration - dark (s): Cumulative duration of long shelter visits during the dark phase
Ttc39 KO mice show a lower long shelter visit duration during the dark phase
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=69&e=10&batch=0&t=&texp=&list=batches&tdose=
3. Key habituation parameters: In general, mice are more active during the first hours of the dark phase on day 1, than during the same period on day 3. To capture these habituation effects, we analyzed the change in activity from day 1 to day 3, i.e., the habituation phase, by taking the ratio of day 3 over day 1.
Activity duration - habituation ratio dark (ratio): Habituation effectChange in cumulative activity number during the dark phase of day 3 compared to the dark phase of day 1
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=84&e=8&batch=0&t=&texp=&list=batches&tdose=
Activity duration - habituation ratio light (ratio): Habituation effectChange in cumulative activity number during the light phase of day 3 compared to the light phase of day 1
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=98&e=8&batch=0&t=&texp=&list=batches&tdose=
4. Key DarkLight index parameter: The light/dark cycle has a strong impact on behavior during each day of the experiment. To assess whether a given behavior is more prominent during the dark or light phase, we defined a LightDark index (dark value/(dark value + light value)). This LightDark index lies between 0.5 and 1.0 if a specific behavior is more pronounced during the dark phase and between 0.0 and 0.5 if it is more prominent during the light phase.
Activity duration - darklight index (index):
The Activity duration - darklight index is lower in the Ttc39 KO mice
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=145&e=7&batch=0&t=&texp=&list=batches&tdose=
5. Key activity pattern parameters: The activity of mice follows a complex circadian pattern, with prominent changes during periods surrounding light/dark phase transition. To capture strain-specific circadian patterns, we studied the behavior of mice by quantifying the anticipation and response to the onset of both light and dark phases.
Activity change in anticipation of dark (fraction of total time): Change in activity during the last 5 hours of the light phase in anticipation of the upcoming dark phase
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=49&e=11&batch=0&t=&texp=&list=batches&tdose=
Activity change in anticipation of light (fraction of total time): Change in activity during the last 5 hours of the dark phase in anticipation of the upcoming light phase
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=50&e=11&batch=0&t=&texp=&list=batches&tdose=
Activity change in response to dark (fraction of total time): Change in activity during the first 2 hours of the dark phase in response to the onset of the dark phase
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=51&e=11&batch=0&t=&texp=&list=batches&tdose=
Activity change in response to light (fraction of total time): Change in activity during the first 2 hours of the light phase in response to the onset of the light phase
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=52&e=11&batch=0&t=&texp=&list=batches&tdose=
6. Key activity bout parameters: Characteristics and quantity of individual activity bouts.
Activity duration - dark (s): Cumulative duration of activity bouts during the dark phase
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=56&e=6&batch=0&t=&texp=&list=batches&tdose=
Activity duration - light (s): Cumulative duration of activity bouts during the light phase
The Ttc39 KO mice show a higher activity duration during the light phase
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=116&e=6&batch=0&t=&texp=&list=batches&tdose=
OnShelter zone number - dark (frequency): Cumulative number of visits to OnShelter zone during the dark phase
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=78&e=6&batch=0&t=&texp=&list=batches&tdose=
Mean activity duration - dark (s): Mean duration per activity bout during the dark phase
No difference
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=57&e=6&batch=0&t=&texp=&list=batches&tdose=
Mean activity duration - light (s): Mean duration per activity bout during the light phase
The Ttc39 KO mice show a higher mean activity duration during the light phase
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=117&e=6&batch=0&t=&texp=&list=batches&tdose=
In addition, there are 28 of the total set of 115 parameters that do not correlate substantially to any of the 20 above mentioned parameters. These 28 'miscellaneous' parameters are in general more difficult to interpret, but may be of interest in specific genotypes or treatments. The entire dataset of this mutant, with p-values of all 115 parameters, can be downloaded here:
http://mousedata.sylics.com/?page=pdfdownloaddata
Cognitive tests in the PhenoTyper
Avoidance learning (shelter task)
The PhenoTyper is equipped with a shelter compartment, with two entrances, in which mice typically spend 80% of their time (resting/sleeping). During the first 4 days, mice develop a preference to enter the shelter through one of the two entrances. The preference index is calculated by : [(number of entries through the preferred entrance) - (number of entries through
non-preferred entrance)]/(total number of entries).
Avoidance learning is studied by automatically applying a mild aversive stimulus (shelter illumination with bright light) during days 5 and 6 each time mice entered the shelter using their preferred entrance, but not when using the other entrance. A reduction in the preference index indicates that a mouse is establishing a specific association between its preferred entrance and the aversive stimulus. During day 7 sanctioning (i.e. shelter illumination) is discontinued, and the stability of the change in preference can be determined.
Avoidance learning is best studied during the Dark phase when shelter illumination is a stronger stimulus than during the light phase. For a detailed explanation see Maroteaux et al. (http://www.ncbi.nlm.nih.gov/pubmed/22846151)
There were no differences between genotypes in the initial preference on Dark 4. The response during Dark 5 and Dark 6 are the same for both WT and KO mice. Both genotypes change their preference. On Dark 7 the WT mice show a stable change in preference. However, the KO mice don’t show a stable change and return to the preferred entrance.
http://mousedata.sylics.com/?page=strainhtp&s=4&htpid=243&e=2&batch=0&t=&texp=&tdose=&list=batches
Appetitive conditioning (pellet task)
During the 4th night, 10 conditioning sessions of 15 minutes are provided in which climbing on top of the shelter is rewarded by delivery of a palatable food pellet in the opposing corner of the cage. Mice can learn two aspects of this task, which are described using two parameters of time:
Reward response over time: If mice learned to associate the drop of a pellet (noise of the pellet dispenser) with the availability of reward, the distance moved from OnShelter to the PelletZone will decrease. This reward response (i.e. the average distance moved between OnShelter to PelletZone) during the 10 x 15 min conditioning sessions can be compared to the day before the task was administered (Q day 3), and the 15 min time bins following 15 min sessions (Q1 after, Q2 after, Q3 after and Q4 after). If mice learned this aspect of the task, they will show a decrease in the average distance moved between OnShelter to PelletZone, specifically during the 15 min when the pellet task is active (Q task) compared to the other Quarters.
Clear learning in all genotypes
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=233&e=3&batch=0&t=&texp=&list=batches&tdose=
Learning effect over time: If mice learned to associate climbing OnShelter with the delivery of the next reward, the distance moved from the PelletZone to OnShelter will decrease. This learning effect (i.e. the average distance moved between PelletZone to OnShelter) during the 10 x 15 min conditioning sessions can be compared to the day before the task was administered (Q day 3), and the 15 min time bins following 15 min sessions (Q1 after, Q2 after, Q3 after and Q4 after). If mice learned this aspect of the task, they will show a decrease in the average distance moved between PelletZone to OnShelter, specifically during the 15 min when the pellet task is active (Q task) compared to the other Quarters.
Clear learning in all genotypes
http://mousedata.sylics.com/?page=strainhtp&loaded=true&s=4&htpid=232&e=3&batch=0&t=&texp=&list=batches&tdose=
Anxiety test (Lightspot avoidance test)
Clear avoidance in both genotypes, no difference between genotypes.
Conventional tests
General assessment
Weight:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=1&e=1&batch=0&t=&texp=&tdose=
Hearing
Startle threshold:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=113&e=10&batch=0&t=&texp=&tdose=
Change in velocity in response to tone before FC:
The Ttc39 KO mice show a lower change in velocity in response to tone before conditioning
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=129&e=9&batch=0&t=&texp=&tdose=
Nociception
Change in velocity in response to electrical foot shock:
There may have been a problem with the shock in the fear conditioning. 4 WT’s do not seem to have received a shock.
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=128&e=9&batch=0&t=&texp=&tdose=
Nesting behavior:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=109&e=12&batch=0&t=&texp=&tdose=
Motor function and activity
Grip Strength
Front paws only:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=7&e=2&batch=0&t=&texp=&tdose=
Front and hind paws together:
Ttc39 KO mice have lower grip strength when both front and hind paws are measured
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=8&e=2&batch=0&t=&texp=&tdose=
Rotarod
Rotations per minute (RPM) reached:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=92&e=8&batch=0&t=&texp=&tdose=
Open Field Total distance moved:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=69&e=6&batch=0&t=&texp=&tdose=
Dark light box Visits to light compartment:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=84&e=7&batch=0&t=&texp=&tdose=
Anxiety
Novel cage induced hypophagia Latency to eat the cracker:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=9&e=3&batch=0&t=&texp=&tdose=
Dark light box
Time spent in the light compartment:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=82&e=7&batch=0&t=&texp=&tdose=
Latency to enter light compartment:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=85&e=7&batch=0&t=&texp=&tdose=
Elevated plus maze
Time spent on open arms:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=56&e=5&batch=0&t=&texp=&tdose=
Visits to open arms (percentage):
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=55&e=5&batch=0&t=&texp=&tdose=
Open Field Time in center:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=72&e=6&batch=0&t=&texp=&tdose=
Learning and memory
T-maze spatial working memory Alternation (percentage):
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=105&e=11&batch=0&t=&texp=&tdose=
Barnes maze spatial reference memory
Learning curves (escape latency):
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=121&e=13&batch=0&t=&texp=&tdose=
Learning curves (distance to target):
On the Barnes Maze, Ttc39 KO mice start off with a much higher distance moved on the first day. After that there is no difference in distance moved during day 2-5.
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=122&e=13&batch=0&t=&texp=&tdose=
Probe trial: hole visits in target octant (probability):
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=125&e=13&batch=0&t=&texp=&tdose=
Contextual fear learning
Freezing duration during baseline, before shock:
Ttc39 KO mice freeze less during the baseline, before they have received a shock.
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=152&e=9&batch=0&t=&texp=&tdose=
Freezing duration (3 s ) in context 1:
No conclusion can be drawn as half of the WT mice probably did not receive a shock
Generalized fear Freezing duration (3 s) context 2, before tone:
No conclusion can be drawn as half of the WT mice probably did not receive a shock
Cued fear learning Freezing duration (3 s) context 2, during tone:
No conclusion can be drawn as half of the WT mice probably did not receive a shock
Object place recognition:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=142&e=4&batch=0&t=&texp=&tdose=
Novel object recognition:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=138&e=4&batch=0&t=&texp=&tdose=
Sensorimotor gating (prepulse inhibition)
ASR curve:
Ttc39 KO mice show a much higher startle response from 95dB onwards
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=102&e=10&batch=0&t=&texp=&tdose=
Prepulse inhibition
PPI at 30 ms interval:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=103&e=10&batch=0&t=&texp=&tdose=
PPI at 100 ms interval:
No difference
http://mousedata.sylics.com/?page=strain&loaded=true&list=batches&s=4<pid=104&e=10&batch=0&t=&texp=&tdose=
Conclusions
The cohort of WT mice in the present study was relatively small and appeared to deviate numerous times from our reference C57BL/6J cohort. For instance, with respect to the activity duration during the light phase, KO mice perfectly resemble the reference set, whereas he group of WT mice substantially deviates.
Thus, all phenotypes of Ttc39 KO mice, such as a higher maximum velocity, lower long shelter visit duration during the dark phase, darklight index and higher mean activity duration during the light phase should be interpreted with care.
With respect to conventional tests, the most pronounced effects in Ttc39 KO mice were a lower grip strength and a much higher startle response in the PPI setup.
Mouse info:
| Mouse ID | Strain | Coat color | Genotype | Ear tag | Internal ID | Sex | Date of Birth | Sub experiment 1 | Sub experiment 2 | Sub experiment 3 |
| PH04646 | Ttc39c | black | WT | L | 3101 | male | 00-00-0000 | |||
| PH04647 | Ttc39c | black | WT | ll | 3116 | male | 00-00-0000 | |||
| PH04648 | Ttc39c | black | WT | r | 3124 | male | 00-00-0000 | |||
| PH04649 | Ttc39c | black | WT | rl | 3132 | male | 00-00-0000 | |||
| PH04650 | Ttc39c | black | WT | rr | 3139 | male | 00-00-0000 | |||
| PH04651 | Ttc39c | black | WT | r | 3142 | male | 00-00-0000 | |||
| PH04652 | Ttc39c | black | WT | ll | 3146 | male | 00-00-0000 | |||
| PH04653 | Ttc39c | black | WT | l | 3160 | male | 00-00-0000 | |||
| PH04654 | Ttc39c | black | WT | r | 3165 | male | 00-00-0000 | |||
| PH04655 | Ttc39c | black | hom | LL | 3104 | male | 00-00-0000 | |||
| PH04656 | Ttc39c | black | hom | l | 3125 | male | 00-00-0000 | |||
| PH04657 | Ttc39c | black | hom | rl | 3126 | male | 00-00-0000 | |||
| PH04658 | Ttc39c | black | hom | ll | 3134 | male | 00-00-0000 | |||
| PH04659 | Ttc39c | black | hom | - | 3135 | male | 00-00-0000 | |||
| PH04660 | Ttc39c | black | hom | ll | 3140 | male | 00-00-0000 | |||
| PH04661 | Ttc39c | black | hom | rl | 3144 | male | 00-00-0000 | |||
| PH04662 | Ttc39c | black | hom | - | 3147 | male | 00-00-0000 | |||
| PH04663 | Ttc39c | black | hom | r | 3148 | male | 00-00-0000 | |||
| PH04664 | Ttc39c | black | hom | - | 3158 | male | 00-00-0000 | |||
| PH04665 | Ttc39c | black | hom | rl | 3161 | male | 00-00-0000 | |||
| PH04666 | Ttc39c | black | hom | ll | 3169 | male | 00-00-0000 | |||
| PH04667 | Ttc39c | black | hom | - | 3170 | male | 00-00-0000 | |||
| PH04668 | Ttc39c | black | hom | r | 3171 | male | 00-00-0000 | |||
| PH04669 | Ttc39c | black | hom | ll | 3175 | male | 00-00-0000 |