Source Paper
Brain-Derived Neurotrophic Factor and Tyrosine Kinase Receptor B Involvement in Amygdala-Dependent Fear Conditioning
Lisa M. Rattiner, Michael Davis, Christopher T. French, Kerry J. Ressler
Journal of Neuroscience • 2004
Pavlovian Fear Conditioning
Objective: Examine the role of BDNF and TrkB in amygdala-dependent learning after a single Pavlovian fear conditioning session
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Equipment2
Materials2
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Protocol Steps
Pavlovian Fear Conditioning
Conduct fear conditioning with paired stimuli to induce learning in subjects
Note: This is the learning session that serves as the basis for examining BDNF and TrkB involvement
View evidence from paper
“the use of Pavlovian fear conditioning as a learning model allows us to examine the concise role of BDNF in the amygdala after a single learning session”
mRNA Level Measurement via In Situ Hybridization
Measure mRNA levels of six different trophic factors (BDNF, neurotrophin 4/5, NGF, NT3, aFGF, and bFGF) at varying time points during the consolidation period after fear conditioning
Note: Temporally specific changes were found only in BDNF gene expression in the basolateral amygdala after paired stimuli supporting learning, but not after exposure to neutral or aversive stimuli alone
View evidence from paper
“Using in situ hybridization, mRNA levels of six different trophic factors [BDNF, neurotrophin (NT) 4/5, NGF, NT3, aFGF, and bFGF) were measured at varying time points during the consolidation period”
Trk Receptor Phosphorylation Analysis via Western Blotting
Measure Trk receptor phosphorylation during the consolidation period following fear conditioning
Note: Increased phosphorylation of Trk receptor was found, suggesting activation of the receptor subsequent to BDNF release
View evidence from paper
“Using Western blotting, we found that the Trk receptor undergoes increased phosphorylation during this consolidation period, suggesting an activation of the receptor subsequent to BDNF release”
K252a Intra-amygdala Infusion
Infuse K252a (Trk receptor antagonist) directly into the amygdala to disrupt neurotrophin signaling
Note: This manipulation disrupted acquisition of fear conditioning, demonstrating the necessity of Trk signaling
View evidence from paper
“disruption of neurotrophin signaling with intra-amygdala infusion of the Trk receptor antagonist K252a disrupted acquisition of fear conditioning”
TrkB.T1 Lentiviral Vector Administration
Administer lentiviral vector expressing dominant-negative TrkB isoform (TrkB.T1) to specifically block TrkB activation in vivo
Note: TrkB.T1 lentivirus blocked fear acquisition without disrupting baseline startle or expression of fear, demonstrating specific role of TrkB in acquisition
View evidence from paper
“In vivo, TrkB.T1 lentivirus blocked fear acquisition without disrupting baseline startle or expression of fear”