Anxiety-Related Behavior Test
Objective: Measured anxiety-related behavior in mice following oxytocin infusion to determine anxiolytic effects and characterize oxytocin receptor expression in serotonergic neurons
This is a Anxiety-Related Behavior Test protocol using mouse as the model organism. The procedure involves 6 procedural steps, 2 equipment items, 2 materials. Extracted from a 2009 paper published in Journal of Neuroscience.
Model and subjects
mouse • oxytocin receptor-reporter mouse (Venus cDNA variant) • unknown • Not specified • Not specified
Study window
Estimated timing pending
Core workflow
Generate oxytocin receptor-reporter mouse model • Examine Venus expression in raphe nuclei • Administer oxytocin infusion
Primary readouts
- Anxiety-related behavior in mice
- Serotonin release within the median raphe nucleus
- Venus expression in raphe nuclei neurons
- Co-localization of oxytocin receptors with serotonergic neurons
Key equipment and reagents
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Protocol Steps
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Generate oxytocin receptor-reporter mouse model
Created transgenic mice in which part of the oxytocin receptor gene was replaced with Venus cDNA (a variant of yellow fluorescent protein) for cellular localization studies
Note: Venus expression used to identify oxytocin receptor-expressing neurons
View evidence from paper
“we generated an oxytocin receptor-reporter mouse in which part of the oxytocin receptor gene was replaced with Venus cDNA”
Examine Venus expression in raphe nuclei
Examined Venus expression to characterize oxytocin receptor-expressing neurons within the raphe nuclei and identify co-localization with serotonergic neurons
Note: About one-half of tryptophan hydroxylase-immunoreactive neurons were positive for Venus, suggesting oxytocin modulation of serotonin release
View evidence from paper
“Examination of the Venus expression revealed that, in the raphe nuclei, about one-half of tryptophan hydroxylase-immunoreactive neurons were positive for Venus”
Administer oxytocin infusion
Infused oxytocin to test effects on serotonin release and anxiety-related behavior
Note: Infusion targeted the median raphe nucleus
View evidence from paper
“Oxytocin infusion facilitated serotonin release within the median raphe nucleus and reduced anxiety-related behavior.”
Measure anxiety-related behavior
Assessed anxiety-related behavior following oxytocin infusion to determine anxiolytic effects
Note: Behavioral testing conducted after oxytocin treatment
View evidence from paper
“reduced anxiety-related behavior”
Administer 5-HT 2A/2C receptor antagonist
Infused 5-HT 2A/2C receptor antagonist to block oxytocin receptor signaling and test mechanism of anxiolytic effects
Note: Used to determine if oxytocin's anxiolytic effects are mediated through serotonergic pathways
View evidence from paper
“Infusion of a 5-HT 2A/2C receptor antagonist blocked the anxiolytic effect of oxytocin”
Measure anxiety-related behavior with antagonist
Assessed anxiety-related behavior following 5-HT 2A/2C receptor antagonist administration to confirm blockade of oxytocin's anxiolytic effects
Note: Behavioral testing conducted to verify mechanism of action
View evidence from paper
“Infusion of a 5-HT 2A/2C receptor antagonist blocked the anxiolytic effect of oxytocin”