Lack of striatal-enriched protein tyrosine phosphatase affected the serotonin system, behavior, and brain morphology in mice methods
Show snapshot details
On this page
This experiment, in seven questions
Jump straight to the part of the recipe you need. Data and provenance labels stay close to the action they support.
Shopping and prep list
What do I need before I start?
Biological model pending
Subject model for the experiment.
- Use
- confirm full cohort details in the source paper
Before you run
What should be confirmed before execution?
First confirmation
Species or subject information is missing.
Confirm before execution
Protocol does not list structured equipment.
Confirm before execution
This page shows passages quoted from the paper. Equipment, materials, times, and temperatures are hidden until the page is rebuilt from the full paper and checked by a reviewer. Check the paper before using this method.
Confirm before execution
Open the source paper before finalizing run-specific details.
Procurement checkpoint
Use source-stated vendors where present. Treat mapped products as sourcing options unless the page marks an exact source match.
Open quote workflowStep-by-step procedure
What do I do, in order?
Social interaction test
The test followed the "resident-intruder" paradigm. A juvenile Balb/c male (4 weeks old) was introduced to the home cage of the tested male. During 10 min, social interactions were registered with the EthoStudio software. Social behavior was evaluated as the total duration of social contacts (intruder's head and body sniffing).
Generation of Ptpn5 knockout mice
In vitro fertilization was performed as described earlier ( ). Oocytes and spermatozoa were taken from C57BL/6 females (4-6 weeks old) and C57BL/6 males (3 months old), respectively. Oocytes were microinjected with a solution containing 100 ng/µL ssODN ( ), 25 ng/µL sgRNA, and an equimolar concentration of Alt-R HiFi Cas9 Nuclease V3 (IDT, Coralville, IA, USA) into the cytoplasm. After the microinjection, embryos were cultivated overnight, and 2-cell stage embryos were transferred into the oviducts of pseudopregnant mothers (females of the CD-1 strain) ( ).
Experimental design
Experiment 2. The behavior of mice was evaluated in the open field (OF) test, in the forced swim test (FST) on the next day, and in the tail suspension test (TST) after 1 day of rest.
Experimental design
Experiment 3. The mice's performance was assessed in the OF, in the Morris water maze (MWM) on the next 5 days, and in the rotarod test after 2 days of rest. After 1 day of rest, the functional activity of the 5-HT 1A receptor was measured; the next day, the functional activity of the 5-HT 2A receptor was studied; and after 2 days of rest, the functional activity of the 5-HT 7 receptor was evaluated.
Elevated plus-maze test
The test was carried out in the apparatus made of gray plastic consisting of four arms connected perpendicularly (closed and open, 30 × 6 cm each) with a central area (6 × 6 cm) ( ). The closed arms were bordered with 20-cm-high walls. The device was elevated by 60 cm above the floor and dimly illuminated with diffuse lighting (100 lx) of a halogen lamp (25 W) placed under the device. The animal's movements were automatically traced with a 3D sensor Kinect 1 connected to the PC through a USB-2 port. During the 5 min of testing, the sensor automatically detected the animal's movement in the open as well as the closed arms. The total path (m), time (%) spent in the center, and open and closed arms were automatically computed by the EthoStudio software. Stretch poses and head dips from the open arms were counted by an experienced rater. The arena was cleaned with we...
Experimental design
Experiment 4. Home cage activity was registered and the operant wall paradigm was carried out over 3 days. Two days later, the animals were euthanized by carbon dioxide asphyxiation followed by decapitation. The frontal cortex, hippocampus, striatum, and midbrain were rapidly dissected, frozen in liquid nitrogen, and stored at -80°C for further assay of monoamine content, TPH2 activity, proteins, and gene expression. The selected brain regions exhibit the most abundant expression of Ptpn5 RNA ( ) and are involved in the 5-HT system regulated processes: mood regulation, decision-making, and depressive and anxiety-related behavior; midbrain is home for 5-HT nuclei.
Behavioral testing
The OF test was carried out to assess the locomotor and exploratory activity. The test took place at the circular arena, 60 cm in diameter ( ), as described earlier ( ). Movement of the mouse was automatically traced for 5 min with a digital camera. The total distance traveled (m) and time (%) spent in the center of the arena were automatically evaluated by the EthoStudio software ( ). The number of vertical postures (rearing) and the number of grooming episodes were marked by experienced rater blinded to the experimental group assignment. After each test, the arena was cleaned with wet (H 2 O 2 ) and dry napkins.
Forced swim test
In the FST and TST, mice were tested for depressive-like behavior. The FST was performed in a clear, cylindrical glass reservoir ( h = 30 cm, d = 15 cm) half filled with water ( T = 25°С) and illuminated from beneath. A mouse was carefully placed into the water for 6 min. The first 2 min of the test are adaptive and were not analyzed. For the latter 4 min, the total immobility time was evaluated by an experienced rater, and depressive-like behavior was determined in correspondence to this parameter.
Measurement outputs
What raw and processed outputs should exist?
Analysis plan
How should the outputs become interpretable results?
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
inferred from protocolPreprocessing / cleaning
Review raw outputs for quality, remove unusable captures, and organize the data into a comparison-ready table or image set.
inferred from protocolScoring or quantification
Quantify the primary endpoint described in the methods before moving into any statistical comparison.
inferred from protocolStatistical comparison
Statistical method not yet structured for this page.
inferred from protocolReporting output
Report representative outputs alongside the summary comparison used to support the conclusion.
inferred from protocolSource and audit
What supports the facts on this page?
Evidence quotes (8)
The test followed the "resident-intruder" paradigm. A juvenile Balb/c male (4 weeks old) was introduced to the home cage of the tested male. During 10 min, social interactions were registered with the EthoStudio software. Social behavior was evaluated as the total duration of social contacts (intruder's head and body sniffing).
In vitro fertilization was performed as described earlier ( ). Oocytes and spermatozoa were taken from C57BL/6 females (4-6 weeks old) and C57BL/6 males (3 months old), respectively. Oocytes were microinjected with a solution containing 100 ng/µL ssODN ( ), 25 ng/µL sgRNA, and an equimolar concentration of Alt-R HiFi Cas9 Nuclease V3 (IDT, Coralville, IA, USA) into the cytoplasm. After the microinjection, embryos were cultivated overnight, and 2-cell stage embryos were transferred into the oviducts of pseudopregnant mothers (females of the CD-1 strain) ( ).
Experiment 2. The behavior of mice was evaluated in the open field (OF) test, in the forced swim test (FST) on the next day, and in the tail suspension test (TST) after 1 day of rest.
Experiment 3. The mice's performance was assessed in the OF, in the Morris water maze (MWM) on the next 5 days, and in the rotarod test after 2 days of rest. After 1 day of rest, the functional activity of the 5-HT 1A receptor was measured; the next day, the functional activity of the 5-HT 2A receptor was studied; and after 2 days of rest, the functional activity of the 5-HT 7 receptor was evaluated.
The test was carried out in the apparatus made of gray plastic consisting of four arms connected perpendicularly (closed and open, 30 × 6 cm each) with a central area (6 × 6 cm) ( ). The closed arms were bordered with 20-cm-high walls. The device was elevated by 60 cm above the floor and dimly illuminated with diffuse lighting (100 lx) of a halogen lamp (25 W) placed under the device. The animal's movements were automatically traced with a 3D sensor Kinect 1 connected to the PC through a USB-2 port. During the 5 min of testing, the sensor automatically detected the animal's movement in the open as well as the closed arms. The total path (m), time (%) spent in the center, and open and closed arms were automatically computed by the EthoStudio software. Stretch poses and head dips from the open arms were counted by an experienced rater. The arena was cleaned with wet (H 2 O 2 ) and dry napkins after each test.
Experiment 4. Home cage activity was registered and the operant wall paradigm was carried out over 3 days. Two days later, the animals were euthanized by carbon dioxide asphyxiation followed by decapitation. The frontal cortex, hippocampus, striatum, and midbrain were rapidly dissected, frozen in liquid nitrogen, and stored at -80°C for further assay of monoamine content, TPH2 activity, proteins, and gene expression. The selected brain regions exhibit the most abundant expression of Ptpn5 RNA ( ) and are involved in the 5-HT system regulated processes: mood regulation, decision-making, and depressive and anxiety-related behavior; midbrain is home for 5-HT nuclei.
The OF test was carried out to assess the locomotor and exploratory activity. The test took place at the circular arena, 60 cm in diameter ( ), as described earlier ( ). Movement of the mouse was automatically traced for 5 min with a digital camera. The total distance traveled (m) and time (%) spent in the center of the arena were automatically evaluated by the EthoStudio software ( ). The number of vertical postures (rearing) and the number of grooming episodes were marked by experienced rater blinded to the experimental group assignment. After each test, the arena was cleaned with wet (H 2 O 2 ) and dry napkins.
In the FST and TST, mice were tested for depressive-like behavior. The FST was performed in a clear, cylindrical glass reservoir ( h = 30 cm, d = 15 cm) half filled with water ( T = 25°С) and illuminated from beneath. A mouse was carefully placed into the water for 6 min. The first 2 min of the test are adaptive and were not analyzed. For the latter 4 min, the total immobility time was evaluated by an experienced rater, and depressive-like behavior was determined in correspondence to this parameter.
Machine-readable layer
[
{
"@context": "https://schema.org",
"@type": "HowTo",
"name": "Lack of striatal-enriched protein tyrosine phosphatase affected the serotonin system, behavior, and brain morphology in mice methods",
"description": "Protocol for Lack of striatal-enriched protein tyrosine phosphatase affected the serotonin system, behavior, and brain morphology in mice methods",
"step": [
{
"@type": "HowToStep",
"position": 1,
"name": "Social interaction test",
"text": "The test followed the \"resident-intruder\" paradigm. A juvenile Balb/c male (4 weeks old) was introduced to the home cage of the tested male. During 10 min, social interactions were registered with the EthoStudio software. Social behavior was evaluated as the total duration of social contacts (intruder's head and body sniffing)."
},
{
"@type": "HowToStep",
"position": 2,
"name": "Generation of Ptpn5 knockout mice",
"text": "In vitro fertilization was performed as described earlier ( ). Oocytes and spermatozoa were taken from C57BL/6 females (4-6 weeks old) and C57BL/6 males (3 months old), respectively. Oocytes were microinjected with a solution containing 100 ng/µL ssODN ( ), 25 ng/µL sgRNA, and an equimolar concentration of Alt-R HiFi Cas9 Nuclease V3 (IDT, Coralville, IA, USA) into the cytoplasm. After the microinjection, embryos were cultivated overnight, and 2-cell stage embryos were transferred into the oviducts of pseudopregnant mothers (females of the CD-1 strain) ( )."
},
{
"@type": "HowToStep",
"position": 3,
"name": "Experimental design",
"text": "Experiment 2. The behavior of mice was evaluated in the open field (OF) test, in the forced swim test (FST) on the next day, and in the tail suspension test (TST) after 1 day of rest."
},
{
"@type": "HowToStep",
"position": 4,
"name": "Experimental design",
"text": "Experiment 3. The mice's performance was assessed in the OF, in the Morris water maze (MWM) on the next 5 days, and in the rotarod test after 2 days of rest. After 1 day of rest, the functional activity of the 5-HT 1A receptor was measured; the next day, the functional activity of the 5-HT 2A receptor was studied; and after 2 days of rest, the functional activity of the 5-HT 7 receptor was evaluated."
},
{
"@type": "HowToStep",
"position": 5,
"name": "Elevated plus-maze test",
"text": "The test was carried out in the apparatus made of gray plastic consisting of four arms connected perpendicularly (closed and open, 30 × 6 cm each) with a central area (6 × 6 cm) ( ). The closed arms were bordered with 20-cm-high walls. The device was elevated by 60 cm above the floor and dimly illuminated with diffuse lighting (100 lx) of a halogen lamp (25 W) placed under the device. The animal's movements were automatically traced with a 3D sensor Kinect 1 connected to the PC through a USB-2 port. During the 5 min of testing, the sensor automatically detected the animal's movement in the open as well as the closed arms. The total path (m), time (%) spent in the center, and open and closed arms were automatically computed by the EthoStudio software. Stretch poses and head dips from the open arms were counted by an experienced rater. The arena was cleaned with we..."
},
{
"@type": "HowToStep",
"position": 6,
"name": "Experimental design",
"text": "Experiment 4. Home cage activity was registered and the operant wall paradigm was carried out over 3 days. Two days later, the animals were euthanized by carbon dioxide asphyxiation followed by decapitation. The frontal cortex, hippocampus, striatum, and midbrain were rapidly dissected, frozen in liquid nitrogen, and stored at -80°C for further assay of monoamine content, TPH2 activity, proteins, and gene expression. The selected brain regions exhibit the most abundant expression of Ptpn5 RNA ( ) and are involved in the 5-HT system regulated processes: mood regulation, decision-making, and depressive and anxiety-related behavior; midbrain is home for 5-HT nuclei."
},
{
"@type": "HowToStep",
"position": 7,
"name": "Behavioral testing",
"text": "The OF test was carried out to assess the locomotor and exploratory activity. The test took place at the circular arena, 60 cm in diameter ( ), as described earlier ( ). Movement of the mouse was automatically traced for 5 min with a digital camera. The total distance traveled (m) and time (%) spent in the center of the arena were automatically evaluated by the EthoStudio software ( ). The number of vertical postures (rearing) and the number of grooming episodes were marked by experienced rater blinded to the experimental group assignment. After each test, the arena was cleaned with wet (H 2 O 2 ) and dry napkins."
},
{
"@type": "HowToStep",
"position": 8,
"name": "Forced swim test",
"text": "In the FST and TST, mice were tested for depressive-like behavior. The FST was performed in a clear, cylindrical glass reservoir ( h = 30 cm, d = 15 cm) half filled with water ( T = 25°С) and illuminated from beneath. A mouse was carefully placed into the water for 6 min. The first 2 min of the test are adaptive and were not analyzed. For the latter 4 min, the total immobility time was evaluated by an experienced rater, and depressive-like behavior was determined in correspondence to this parameter."
}
],
"tool": [],
"supply": [],
"isBasedOn": {
"@type": "ScholarlyArticle",
"headline": "Lack of striatal-enriched protein tyrosine phosphatase affected the serotonin system, behavior, and brain morphology in mice",
"datePublished": "2026",
"author": [
{
"@type": "Person",
"name": "Vitalii Moskaliuk"
},
{
"@type": "Person",
"name": "Polina Komleva"
},
{
"@type": "Person",
"name": "Nikita Khotskin"
},
{
"@type": "Person",
"name": "Alla Arefieva"
},
{
"@type": "Person",
"name": "Oleg Shevelev"
},
{
"@type": "Person",
"name": "Alexey Korablev"
},
{
"@type": "Person",
"name": "Irina Serova"
},
{
"@type": "Person",
"name": "Nariman Battulin"
},
{
"@type": "Person",
"name": "Alexander Kulikov"
},
{
"@type": "Person",
"name": "Vladimir Naumenko"
},
{
"@type": "Person",
"name": "Darya Bazovkina"
},
{
"@type": "Person",
"name": "Elizabeth Kulikova"
}
],
"identifier": "10.3389/fpsyt.2025.1730197"
}
},
{
"@context": "https://schema.org",
"@type": "BreadcrumbList",
"itemListElement": [
{
"@type": "ListItem",
"position": 1,
"name": "Experiments",
"item": "https://replicatescience.com/experiments"
},
{
"@type": "ListItem",
"position": 2,
"name": "Lack of striatal-enriched protein tyrosine phosphatase affected the serotonin system, behavior, and brain morphology in mice methods",
"item": "https://replicatescience.com/experiments/lack-of-striatal-enriched-protein-tyrosine-phosphatase-affected-the-serotonin-system-behavior-and-brain-morphology-in-mice-methods-vitalii-moskaliuk-pmc12848919/lack-of-striatal-enriched-protein-tyrosine-phosphatase-affected-the-serotonin-sy"
}
]
}
]