Conditioned Fear Test
Objective: Measurement of exaggerated conditioned fear responses induced by uncontrollable stress, and evaluation of how freewheel running prevents learned helplessness behaviors including poor escape responding and expression of exaggerated conditioned fear
This is a Conditioned Fear Test protocol using rat as the model organism. The procedure involves 6 procedural steps, 2 equipment items. Extracted from a 2003 paper published in Journal of Neuroscience.
Model and subjects
rat • Not specified • unknown • Not specified • Not specified
Study window
~6 week study window
Core workflow
Freewheel running intervention • Uncontrollable tail shock exposure • Shuttle box escape test
Primary readouts
- Shuttle box escape deficit (measure of learned helplessness)
- Exaggerated conditioned fear responses
- Tail shock-induced activity of dorsal raphe serotonergic neurons (via c-Fos/5-HT immunohistochemistry)
- Basal 5-HT1A inhibitory autoreceptor mRNA expression in rostral-mid dorsal raphe nucleus
Key equipment and reagents
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Protocol Steps
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Freewheel running intervention
Rats were provided with voluntary access to freewheel running for 6 weeks prior to stress exposure
Note: This intervention was compared against sedentary control rats
View evidence from paper
“The current study determined the effects of 6 weeks of voluntary freewheel running on LH behaviors”
Uncontrollable tail shock exposure
Rats were exposed to uncontrollable tail shock to induce learned helplessness and exaggerated conditioned fear responses
Note: This stress manipulation was used to induce learned helplessness behaviors
View evidence from paper
“the exaggerated conditioned fear that is induced by uncontrollable tail shock in sedentary rats”
Shuttle box escape test
Assessment of escape responding in the shuttle box apparatus following uncontrollable stress exposure
Note: Measures the shuttle box escape deficit as an indicator of learned helplessness
View evidence from paper
“Freewheel running prevented the shuttle box escape deficit and the exaggerated conditioned fear”
Conditioned fear test
Measurement of exaggerated conditioned fear responses following uncontrollable stress exposure
Note: Assesses the expression of exaggerated conditioned fear as a learned helplessness behavior
View evidence from paper
“Freewheel running prevented the shuttle box escape deficit and the exaggerated conditioned fear that is induced by uncontrollable tail shock”
Double c-Fos/5-HT immunohistochemistry
Neural activity assessment in dorsal raphe nucleus serotonergic neurons following tail shock stress
Note: Reveals tail shock-induced activity of 5-HT neurons in the rostral-mid DRN
View evidence from paper
“double c-Fos/5-HT immunohistochemistry revealed that physical activity attenuated tail shock-induced activity of 5-HT neurons in the rostral–mid DRN”
5-HT1A autoreceptor mRNA expression analysis
Assessment of basal 5-HT1A inhibitory autoreceptor mRNA expression in the dorsal raphe nucleus
Note: Measures changes in 5-HT1A autoreceptor expression following 6 weeks of freewheel running
View evidence from paper
“Six weeks of freewheel running also resulted in a basal increase in 5-HT1A inhibitory autoreceptor mRNA in the rostral–mid DRN”