Hippocampal Lesion Control Study
Objective: Compare behavioral performance and hippocampal LTP between D-AP5-treated rats and rats with bilateral hippocampal lesions induced by ibotenic acid to establish that NMDA receptor activation is necessary for spatial learning
This is a Hippocampal Lesion Control Study protocol using rat as the model organism. The procedure involves 8 procedural steps, 4 equipment items, 4 materials. Extracted from a 1992 paper published in Journal of Neuroscience.
Model and subjects
rat • Not specified • unknown • Not specified • Not specified
Study window
Estimated timing pending
Core workflow
Surgical implantation of infusion system • Spatial learning training • Anesthesia induction
Primary readouts
- Spatial learning performance in water maze task
- Hippocampal LTP magnitude and inducibility
- Extracellular D-AP5 concentration in hippocampus via microdialysis
- Whole tissue D-AP5 concentration
Key equipment and reagents
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Protocol Steps
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Surgical implantation of infusion system
Rats receive chronic infusion of D-AP5 into the lateral ventricle via osmotic minipumps at concentrations ranging from 0-50 mM
Note: Bilateral hippocampal lesions induced by ibotenic acid in control group
View evidence from paper
“Rats were chronically infused with D-AP5 into the lateral ventricle at a range of concentrations (0–50 mM) via osmotic minipumps”
Spatial learning training
Rats trained to find and escape onto a hidden platform in an open-field water maze task
Note: Behavioral learning phase occurs before electrophysiology
View evidence from paper
“They were first trained to find and escape onto a hidden platform in an open-field water maze task”
Anesthesia induction
After behavioral learning, rats are anesthetized with urethane
Note: Preparation for electrophysiology and sampling
View evidence from paper
“After the behavioral learning, they were anesthetized with urethane”
Hippocampal LTP evocation and monitoring
Attempt to evoke and monitor hippocampal long-term potentiation in anesthetized rats
Note: Electrophysiological recording phase
View evidence from paper
“an attempt was made to evoke and monitor hippocampal LTP”
Microdialysis sampling
Extracellular samples of D-AP5 collected from hippocampus using microdialysis
Note: Concurrent with or following LTP monitoring
View evidence from paper
“Extracellular samples of D-AP5 in hippocampus were then taken using microdialysis”
Tissue collection
Animals killed and tissue samples dissected from hippocampus
Note: Final step before chemical analysis
View evidence from paper
“finally, the animals were killed and tissue samples dissected”
Chemical analysis of D-AP5
Microdialysis and tissue samples analyzed for D-AP5 content using HPLC with fluorescence detection
Note: Determines extracellular and tissue concentrations of D-AP5
View evidence from paper
“The microdialysis and tissue samples were analyzed for D-AP5 content using HPLC with fluorescence detection”
EGTA infusion control experiment
Infusion of 20 mM EGTA to test calcium-dependent D-AP5 trapping mechanism
Note: Separate control experiment to investigate D-AP5 accessibility
View evidence from paper
“Infusion of 20 mM EGTA was found to cause a sevenfold increase in D-AP5 in the dialysis perfusates”