Home › Neuroscience › Investigation of Synaptic Tagging/Capture and Cross-capture using Acute Hippocampal Slices from Rodents
Neuroscience JoVE (Open Access) Citable · DOI

Investigation of Synaptic Tagging/Capture and Cross-capture using Acute Hippocampal Slices from Rodents

DOI: 10.3791/53008-v
What you'll learn
  • ✓ Prepare acute hippocampal slices and maintain viable CA1 pyramidal neurons in vitro
  • ✓ Record field-potential synaptic responses using extracellular electrode techniques
  • ✓ Induce and characterize long-term potentiation/depression and synaptic tagging mechanisms
  • ✓ Investigate associative plasticity processes including synaptic capture and cross-capture
Protocol

This video article describes experimental procedures to study long-term plasticity and its associative processes such as synaptic tagging, capture and cross-tagging in the CA1 pyramidal neurons using acute hippocampal slices from rodents.

Difficulty
advanced
Total time
~4–6 hours per experiment (slice preparation ~1.5 hrs; recording/induction protocols ~2–3 hrs per slice)
Model organism
Rodent (mouse/rat) acute hippocampal slices
Biosafety
BSL-1

Steps

1
Prepare artificial cerebrospinal fluid and interface chamber

Mix ACSF to physiological ionic composition and equilibrate with carbogen gas. Set up the interface-type recording chamber with appropriate heating and perfusion systems for slice viability.

▶ 01:53
2
Prepare acute hippocampal slices from rodent brain

Extract and rapidly section hippocampus into 400 μm transverse slices using a vibratome. Recover slices in oxygenated ACSF at appropriate temperature before recording.

▶ 02:52
3
Record CA3-CA1 synaptic responses using field electrodes

Position extracellular recording and stimulation electrodes in the CA1 stratum radiatum to measure field excitatory postsynaptic potentials (fEPSPs) evoked by Schaffer collateral stimulation.

▶ 06:36
4
Induce long-term potentiation and depression protocols

Apply high-frequency or low-frequency stimulation patterns to the Schaffer collateral-CA1 pathway to induce LTP or LTD, and monitor resulting changes in synaptic strength over time.

▶ 08:13
5
Analyze transverse slice anatomy and electrode positioning

Visualize hippocampal CA1 and CA3 regions and confirm correct electrode placement within stratum radiatum for reliable field-potential recording.

▶ 10:41
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