P301S Mutant Human Tau Transgenic Mice Manifest Early Symptoms of Human Tauopathies with Dementia and Altered Sensorimotor Gating methods
Aim. Evidence-backed execution summary for P301S Mutant Human Tau Transgenic Mice Manifest Early Symptoms of Human Tauopathies with Dementia and Altered Sensorimotor Gating methods from P301S Mutant Human Tau Transgenic Mice Manifest Early Symptoms of Human Tauopathies with Dementia and Altered Sensorimotor Gating.
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mouse
Subject model for the experiment.
- Use
- confirm full cohort details in the source paper
Immunohistochemistry
reagent used in the protocol.
- Use
- To evaluate the affected brain region in regard to sensorimotor gating, we assessed male tauopathy model mice at 4 months of age. Mice under deep pentobarbital anesthesia were perfused via the aorta with 50 mL of phosphate-buffered saline (PBS). After perfusion, the brain was quickly removed, followed by fixation wi...
Immunoblotting
reagent used in the protocol.
- Use
- Tau protein levels of brains of mice at 6 months of age were determined by homogenizing brains (whole cortex and whole hippocampus of each mouse) in 2 ml/g tissue of ice-cold high-salt reassembly buffer (RAB-HS) (0.1 M MES, 1 mM EGTA, 0.5 mM MgSO4, 0.75 M NaCl, 0.02 M NaF, 1 mM PMSF, and protease inhibitor cocktail...
Experimental design
Six-month-old female animals with a B6C3H/F1 background (tauopathy model mice: n = 29; littermate wild type mice: n = 11) were used to perform the Morris water maze test. All other studied animals were male with a C57BL/6J background, and in each individual test, tauopathy model mice (n...
- Use
- Six-month-old female animals with a B6C3H/F1 background (tauopathy model mice: n = 29; littermate wild type mice: n = 11) were used to perform the Morris water maze test. All other studied animals were male with a C57BL/6J background, and in each individual test, tauopathy model mice (n...
Impaired spontaneous alternation in tauopathy model mice
We performed the Y-maze test to evaluate spontaneous alternative behavior. Tauopathy model mice demonstrated increased numbers of entries into each arm, total alternations, and total distance traveled ( ), reflecting hyperactivity. However, the percentage of alternations was significantly decreased ( ).
- Use
- We performed the Y-maze test to evaluate spontaneous alternative behavior. Tauopathy model mice demonstrated increased numbers of entries into each arm, total alternations, and total distance traveled ( ), reflecting hyperactivity. However, the percentage of alternations was significantly decreased ( ).
No significant differences in light/dark transition test
In the light/dark transition test, no statistically significant differences were observed between tauopathy model mice and wild type mice ( ). However, latency to enter the light chamber tended to be decreased in the former ( ).
- Use
- In the light/dark transition test, no statistically significant differences were observed between tauopathy model mice and wild type mice ( ). However, latency to enter the light chamber tended to be decreased in the former ( ).
Increased hyperactivity and decreased anxiety-like behavior in tauopathy model mice
We then applied the open field test paradigm to explore anxiety-like behavior ( ). Tauopathy model mice showed an increasing tendency in total locomotive distance ( ), and vertical activity was increased in the latter half of the test ( ). Time spent in the center of the field was also increased ( ). Stereotypic loc...
- Use
- We then applied the open field test paradigm to explore anxiety-like behavior ( ). Tauopathy model mice showed an increasing tendency in total locomotive distance ( ), and vertical activity was increased in the latter half of the test ( ). Time spent in the center of the field was also increased ( ). Stereotypic loc...
Locomotor activity and motor coordination
In the rotarod test, tauopathy model mice demonstrated no significant change in latencies to fall compared with wild type mice ( ). Since the effect of motor learning reached a plateau in wild type mice after the 4 th trial, we also analyzed the performance of each genotype in the 4 th -6 th trials. In these trials,...
- Use
- In the rotarod test, tauopathy model mice demonstrated no significant change in latencies to fall compared with wild type mice ( ). Since the effect of motor learning reached a plateau in wild type mice after the 4 th trial, we also analyzed the performance of each genotype in the 4 th -6 th trials. In these trials,...
Decreased sociability/object recognition memory in tauopathy model mice
During the social interaction test in a one-chamber novel environment, no statistical significance was observed in the time spent and the number of contacts, duration of active contacts, mean of duration/contact ratio, and distance traveled between the two groups of mice ( ).
- Use
- During the social interaction test in a one-chamber novel environment, no statistical significance was observed in the time spent and the number of contacts, duration of active contacts, mean of duration/contact ratio, and distance traveled between the two groups of mice ( ).
Decreased sociability/object recognition memory in tauopathy model mice
Crawley's three-chamber social approach test consists of a sociability test and a social novelty preference test. These tests assess social interaction that is relatively independent of locomotor activity compared to other social interaction tests, because the preference of the mice can be quantified based on the ti...
- Use
- Crawley's three-chamber social approach test consists of a sociability test and a social novelty preference test. These tests assess social interaction that is relatively independent of locomotor activity compared to other social interaction tests, because the preference of the mice can be quantified based on the ti...
Decreased sociability/object recognition memory in tauopathy model mice
The 1 st trial with one-stranger mouse (A-D, I, J), the 2 nd trial with one-familiar mouse and one-stranger mouse (E-H, K, L). Time spent in each chamber was tested with one-way ANOVA, followed by Fisher's least significant difference method as post hoc analysis within groups, with Student's t-test betwe...
- Use
- The 1 st trial with one-stranger mouse (A-D, I, J), the 2 nd trial with one-familiar mouse and one-stranger mouse (E-H, K, L). Time spent in each chamber was tested with one-way ANOVA, followed by Fisher's least significant difference method as post hoc analysis within groups, with Student's t-test betwe...
Image and data analysis
Software used for acquisition, scoring, statistics, or reporting.
- Use
- The applications used for the behavioral studies (Image LD, Image EP, Image PS, Image SI, Image YM, Image CSI, Image BM, Image FZ, and Image TS) were based on the public domain NIH Image program (developed at the U.S. National Institutes of Health available at http://rsb.info.nih.gov/nih-image/ ) and the ImageJ prog...
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Sensorimotor gating
The prepulse inhibition (PPI) test is widely used to measure deficits in information-processing abilities or sensorimotor gating in schizophrenia patients, and it can be employed in both human and animal experiments. PPI is defined as the degree (%) by which the motor acoustic startle response is reduced when the startle-eliciting stimulus is preceded by a brief, low-intensity, non-eliciting stimulus. Tauopathy model mice had lower startle amplitudes than wild type mice at both 110 dB and 120 dB, and this was statistically significant at 120 dB ( ). Especially, the %PPI, an index of sensorimotor gating, was significantly greater in tauopathy model mice ( ). The reduced PPI values at 120 dB rather than those at 110 dB might be a ceiling effect caused by the strong intensity of the startle stimulus.
Decreased sociability/object recognition memory in tauopathy model mice
During the social interaction test in a one-chamber novel environment, no statistical significance was observed in the time spent and the number of contacts, duration of active contacts, mean of duration/contact ratio, and distance traveled between the two groups of mice ( ).
Open field test
Locomotor activity was measured using the open field test, which was performed with 14-week-old male mice. Each mouse was placed in the center of the open field apparatus (40 × 40 × 30 cm; Accuscan Instruments, Columbus, OH, USA). Total distance traveled (in cm), vertical activity (rearing measured by counting the number of photobeam interruptions), time spent in the center, beam-break counts for stereotyped behaviors, and number of fecal boli were recorded. Data were collected for 120 min.
Light/dark transition test
Light/dark transition test was performed as previously described. The apparatus used consisted of a cage (21 × 42 × 25 cm) divided into two chambers of equal size by a partition containing a door (O'Hara & Co.). One chamber was brightly illuminated (390 lux), whereas the other was dark (2 lux). Mice were placed into the dark side and allowed to move freely between the two chambers with the door open for 10 min. The total number of transitions between chambers, time spent in each side, first latency to enter the light side, and distance traveled were recorded automatically using Image LD software (see 'Image and data analysis').
Elevated plus-maze test
Elevated plus-maze test was performed as previously described. The elevated plus-maze (O'Hara & Co.) consisted of two open arms (25 × 5 cm) and two enclosed arms of the same size, with 15-cm high transparent walls. The arms and central square were made of white plastic plates and were elevated to a height of 55 cm above the floor. To minimize the likelihood of animals falling from the apparatus, 3-mm high plastic ledges were provided for the open arms. Arms of the same type were arranged at opposite sides to each other. Each mouse was placed in the central square of the maze (5 × 5 cm), facing one of the closed arms. Mouse behavior was recorded during a 10-min test period. The numbers of entries into, and the time spent in open and enclosed arms, were recorded. The illumination level was 100 lux at the center of the maze. For data analysis, we used the following four measures:...
Porsolt forced swim test
The Porsolt forced swim test apparatus consisted of four Plexiglas cylinders (20 cm high × 10 cm diameter). A nontransparent panel separated the cylinders to prevent the mice from seeing each other (O'Hara & Co.). The cylinders were filled with water (23°C) up to a height of 7.5 cm. Mice were placed into the cylinders, and their behavior was recorded over a 10-min test period (Day 1). Retention tests were administered 24 hours after training (Day 2). Data acquisition and analysis were performed automatically using Image PS software (see 'Image and data analysis').
Social interaction test in a novel environment (one-chamber social interaction test)
In the social interaction test, two mice of identical genotypes that were previously housed in different cages were placed in a box together (40 × 40 × 30 cm) and allowed to explore freely for 10 min. Social behavior was monitored with a CCD camera connected to a computer. Analysis was performed automatically using Image SI software (see 'Image and data analysis'). The total number of contacts, total duration of active contacts, total contact duration, mean duration per contact, and total distance traveled were measured. Active contact was defined as follows. Images were captured at 1 frame per second, and distance traveled between two successive frames was calculated for each mouse. If the two mice contacted each other and the distance traveled by either mouse was longer than 2 cm, the behavior was considered as 'active contact'. Genotypic mismatch was f...
Startle response/Prepulse inhibition (PPI) test
An acoustic startle reflex measurement system was used (O'Hara & Co.). A test session was begun by placing a 16-week-old mouse in a Plexiglas cylinder, where it was left undisturbed for 10 min. The duration of white noise, used as the startle stimulus, was 40 ms for all trial types. The startle response was recorded for 140 ms (measuring response every 1 ms) starting with the onset of the prepulse stimulus. The background noise level in each chamber was 70 dB. The peak startle amplitude recorded during the 140-ms sampling window was used as dependent variable. A test session consisted of 6 trial types (i.e., two types for startle stimulus-only trials, and four types for PPI trials). Intensity of the acoustic startle stimulus was 110 or 120 dB. The prepulse sound was presented 100 ms before the startle stimulus, and its intensity was 74 or 78 dB. Four combinations of prepulse and start...
Measurement outputs
What raw and processed outputs should exist?
We performed the Y-maze test to evaluate spontaneous alternative behavior. Tauopathy model mice demonstrated increased numbers of entries into each arm, total alternations, and...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
In the light/dark transition test, no statistically significant differences were observed between tauopathy model mice and wild type mice ( ). However, latency to enter the ligh...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
We then applied the open field test paradigm to explore anxiety-like behavior ( ). Tauopathy model mice showed an increasing tendency in total locomotive distance ( ), and verti...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
Total distance traveled (A), vertical activity (B), center time (C), stereotypic counts (D). Tauopathy model mice traveled longer distance ( F (1, 33) = 4.467, p ...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
Analysis plan
How should the outputs become interpretable results?
Acquisition
Collect the raw assay or blot output and retain identifiers for each sample and experimental group.
inferred from protocolPreprocessing / cleaning
Tail suspension test.
from paperScoring or quantification
Quantify the primary readouts for this experiment: We performed the Y-maze test to evaluate spontaneous alternative behavior. Tauopathy model mice demonstrated increased numbers of entries into each arm, total alternations, and...; In the light/dark transition test, no statistically significant differences were observed between tauopathy model mice and wild type mice ( ). However, latency to enter the ligh...; We then applied the open field test paradigm to explore anxiety-like behavior ( ). Tauopathy model mice showed an increasing tendency in total locomotive distance ( ), and verti...; Total distance traveled (A), vertical activity (B), center time (C), stereotypic counts (D). Tauopathy model mice traveled longer distance ( F (1, 33) = 4.467, p ....
from paperNormalization
Normalize expression or signal values against the stated control or loading reference before comparing groups.
inferred from protocolStatistical comparison
Tail suspension test. One wild type mouse dropped during the test and failed to complete the task. No statistical significance was observed between tauopathy model mice and wild...; In the light/dark transition test, no statistically significant differences were observed between tauopathy model mice and wild type mice ( ). However, latency to enter the ligh...; Total distance traveled (A), vertical activity (B), center time (C), stereotypic counts (D). Tauopathy model mice traveled longer distance ( F (1, 33) = 4.467, p ...; In the rotarod test, tauopathy model mice demonstrated no significant change in latencies to fall compared with wild type mice ( ). Since the effect of motor learning reached a...
from paperReporting output
Report representative outputs alongside summary comparisons for We performed the Y-maze test to evaluate spontaneous alternative behavior. Tauopathy model mice demonstrated increased numbers of entries into each arm, total alternations, and..., In the light/dark transition test, no statistically significant differences were observed between tauopathy model mice and wild type mice ( ). However, latency to enter the ligh..., We then applied the open field test paradigm to explore anxiety-like behavior ( ). Tauopathy model mice showed an increasing tendency in total locomotive distance ( ), and verti..., Total distance traveled (A), vertical activity (B), center time (C), stereotypic counts (D). Tauopathy model mice traveled longer distance ( F (1, 33) = 4.467, p ....
inferred from protocolStructured statistical methods
Tail suspension test. One wild type mouse dropped during the test and failed to complete the task. No statistical significance was observed between tauopathy model mice and wild...; In the light/dark transition test, no statistically significant differences were observed between tauopathy model mice and wild type mice ( ). However, latency to enter the ligh...; Total distance traveled (A), vertical activity (B), center time (C), stereotypic counts (D). Tauopathy model mice traveled longer distance ( F (1, 33) = 4.467, p ...; In the rotarod test, tauopathy model mice demonstrated no significant change in latencies to fall compared with wild type mice ( ). Since the effect of motor learning reached a...
source structuredSource and audit
What supports the facts on this page?
Evidence quotes (8)
The prepulse inhibition (PPI) test is widely used to measure deficits in information-processing abilities or sensorimotor gating in schizophrenia patients, and it can be employed in both human and animal experiments. PPI is defined as the degree (%) by which the motor acoustic startle response is reduced when the startle-eliciting stimulus is preceded by a brief, low-intensity, non-eliciting stimulus. Tauopathy model mice had lower startle amplitudes than wild type mice at both 110 dB and 120 dB, and this was statistically significant at 120 dB ( ). Especially, the %PPI, an index of sensorimotor gating, was significantly greater in tauopathy model mice ( ). The reduced PPI values at 120 dB rather than those at 110 dB might be a ceiling effect caused by the strong intensity of the startle stimulus.
During the social interaction test in a one-chamber novel environment, no statistical significance was observed in the time spent and the number of contacts, duration of active contacts, mean of duration/contact ratio, and distance traveled between the two groups of mice ( ).
Locomotor activity was measured using the open field test, which was performed with 14-week-old male mice. Each mouse was placed in the center of the open field apparatus (40 × 40 × 30 cm; Accuscan Instruments, Columbus, OH, USA). Total distance traveled (in cm), vertical activity (rearing measured by counting the number of photobeam interruptions), time spent in the center, beam-break counts for stereotyped behaviors, and number of fecal boli were recorded. Data were collected for 120 min.
Light/dark transition test was performed as previously described. The apparatus used consisted of a cage (21 × 42 × 25 cm) divided into two chambers of equal size by a partition containing a door (O'Hara & Co.). One chamber was brightly illuminated (390 lux), whereas the other was dark (2 lux). Mice were placed into the dark side and allowed to move freely between the two chambers with the door open for 10 min. The total number of transitions between chambers, time spent in each side, first latency to enter the light side, and distance traveled were recorded automatically using Image LD software (see 'Image and data analysis').
Elevated plus-maze test was performed as previously described. The elevated plus-maze (O'Hara & Co.) consisted of two open arms (25 × 5 cm) and two enclosed arms of the same size, with 15-cm high transparent walls. The arms and central square were made of white plastic plates and were elevated to a height of 55 cm above the floor. To minimize the likelihood of animals falling from the apparatus, 3-mm high plastic ledges were provided for the open arms. Arms of the same type were arranged at opposite sides to each other. Each mouse was placed in the central square of the maze (5 × 5 cm), facing one of the closed arms. Mouse behavior was recorded during a 10-min test period. The numbers of entries into, and the time spent in open and enclosed arms, were recorded. The illumination level was 100 lux at the center of the maze. For data analysis, we used the following four measures: the percentage of entries into the open arms, time spent in the open arms (sec), number of total entries, and total distance traveled (cm). Data acquisition and analysis were performed automatically using Image EP software (see 'Image and data analysis'). Three wild type mice and a tauop...
The Porsolt forced swim test apparatus consisted of four Plexiglas cylinders (20 cm high × 10 cm diameter). A nontransparent panel separated the cylinders to prevent the mice from seeing each other (O'Hara & Co.). The cylinders were filled with water (23°C) up to a height of 7.5 cm. Mice were placed into the cylinders, and their behavior was recorded over a 10-min test period (Day 1). Retention tests were administered 24 hours after training (Day 2). Data acquisition and analysis were performed automatically using Image PS software (see 'Image and data analysis').
In the social interaction test, two mice of identical genotypes that were previously housed in different cages were placed in a box together (40 × 40 × 30 cm) and allowed to explore freely for 10 min. Social behavior was monitored with a CCD camera connected to a computer. Analysis was performed automatically using Image SI software (see 'Image and data analysis'). The total number of contacts, total duration of active contacts, total contact duration, mean duration per contact, and total distance traveled were measured. Active contact was defined as follows. Images were captured at 1 frame per second, and distance traveled between two successive frames was calculated for each mouse. If the two mice contacted each other and the distance traveled by either mouse was longer than 2 cm, the behavior was considered as 'active contact'. Genotypic mismatch was found in one pair (one tauopathy model pair), and they could not be analyzed.
An acoustic startle reflex measurement system was used (O'Hara & Co.). A test session was begun by placing a 16-week-old mouse in a Plexiglas cylinder, where it was left undisturbed for 10 min. The duration of white noise, used as the startle stimulus, was 40 ms for all trial types. The startle response was recorded for 140 ms (measuring response every 1 ms) starting with the onset of the prepulse stimulus. The background noise level in each chamber was 70 dB. The peak startle amplitude recorded during the 140-ms sampling window was used as dependent variable. A test session consisted of 6 trial types (i.e., two types for startle stimulus-only trials, and four types for PPI trials). Intensity of the acoustic startle stimulus was 110 or 120 dB. The prepulse sound was presented 100 ms before the startle stimulus, and its intensity was 74 or 78 dB. Four combinations of prepulse and startle stimuli were employed (74-110 dB, 78-110 dB, 74-120 dB, and 78-120 dB). Six blocks of the 6 trial types were presented in a pseudorandom order such that each trial type was presented once within a block. The average inter-trial interval was 15 sec (range, 10-20 sec)....
Machine-readable layer
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"name": "P301S Mutant Human Tau Transgenic Mice Manifest Early Symptoms of Human Tauopathies with Dementia and Altered Sensorimotor Gating methods",
"item": "https://replicatescience.com/experiments/p301s-mutant-human-tau-transgenic-mice-manifest-early-symptoms-of-human-tauopathies-with-dementia-and-altered-sensorimotor-gating-methods-hiroki-takeuchi-pmc3115982/p301s-mutant-human-tau-transgenic-mice-manifest-early-symptoms-of-human-tauopathies-with-dementia-an-mlph47wi"
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