Transgenic PKA-Deficient Mouse Analysis
Objective: Define the time course of requirement for protein kinase A (PKA) and protein synthesis in long-term memory for contextual fear conditioning, and determine whether contextual memory has one or two brief consolidation periods depending on training intensity
This is a Transgenic PKA-Deficient Mouse Analysis protocol using mouse as the model organism. The procedure involves 5 procedural steps, 2 materials. Extracted from a 1998 paper published in Learning & Memory.
Model and subjects
mouse • transgenic mice expressing R(AB) • not specified • not specified • not specified • not specified
Study window
~3 hour study window
Core workflow
Contextual Fear Conditioning Training • Injection of Protein Synthesis Inhibitor • Injection of PKA Inhibitor
Primary readouts
- Time course of amnesia in transgenic mice with reduced PKA activity
- Number and timing of critical consolidation periods (one vs. two)
- Sensitivity to protein synthesis inhibitors at different post-training intervals
- Sensitivity to PKA inhibitors at different post-training intervals
Key equipment and reagents
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Protocol Steps
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Contextual Fear Conditioning Training
Mice undergo contextual fear conditioning training with either weak or strong training protocols
Note: Different training procedures recruit either one or two critical periods for memory consolidation
View evidence from paper
“Different Training Procedures Recruit Either One or Two Critical Periods for Contextual Memory Consolidation”
Injection of Protein Synthesis Inhibitor
Anisomycin (protein synthesis inhibitor) is injected at various times after training to determine sensitivity periods
Note: Timing varies to map consolidation periods
View evidence from paper
“injecting the protein synthesis inhibitor anisomycin or the PKA inhibitor Rp-cAMPs at various times after training”
Injection of PKA Inhibitor
Rp-cAMPs (PKA inhibitor) is injected at various times after training to determine sensitivity periods
Note: Timing varies to map consolidation periods
View evidence from paper
“injecting the protein synthesis inhibitor anisomycin or the PKA inhibitor Rp-cAMPs at various times after training”
Assessment of Memory Consolidation Timeline
Evaluate amnesia time course in transgenic mice with genetically reduced PKA activity in hippocampus and compare to pharmacologically inhibited mice
Note: PKA- and protein synthesis-dependent memory develops between 1 hr and 3 hr after training
View evidence from paper
“This PKA- and protein synthesis-dependent memory develops between 1 hr and 3 hr after training”
Differentiate Training Intensity Effects
Compare consolidation periods between weak and strong training protocols
Note: Weak training shows two time periods of sensitivity; stronger training exhibits only one
View evidence from paper
“Weak training shows two time periods of sensitivity to inhibitors of protein synthesis and PKA, whereas stronger training exhibits only one”