Home › Neuroscience › The Use of Trace Eyeblink Classical Conditioning to Assess Hippocampal Dysfunction in a Rat Model of Fetal Alcohol Spectrum Disorders
Neuroscience JoVE (Open Access) Citable · DOI

The Use of Trace Eyeblink Classical Conditioning to Assess Hippocampal Dysfunction in a Rat Model of Fetal Alcohol Spectrum Disorders

DOI: 10.3791/55350-v
What you'll learn
  • ✓ Perform neonatal alcohol exposure and eyelid surgery in rats
  • ✓ Conduct trace eyeblink classical conditioning assessment protocols
  • ✓ Analyze hippocampal-dependent associative learning deficits
  • ✓ Interpret adaptive conditioned response development across training sessions
Protocol

Trace eyeblink classical conditioning (ECC) was used to assess hippocampal-dependent associative learning in adult rats that were administered a high concentration (11.9% v/v) of alcohol during early neonatal brain development. In general, ECC procedures are sound diagnostic tools for detecting brain dysfunction across many psychological and biomedical settings.

Difficulty
advanced
Total time
~8-10 weeks (neonatal exposure + recovery + surgical implantation + 6 conditioning sessions over 2-3 weeks)
Model organism
Rat (neonatal and adult)
Biosafety
BSL-1

Steps

1
Administer neonatal alcohol exposure

Treat neonatal rats with high-concentration ethanol (11.9% v/v) during early developmental stages to induce fetal alcohol spectrum disorder phenotype. Allow animals to mature to adulthood before behavioral testing.

▶ 01:02
2
Prepare materials and surgical equipment

Gather and sterilize all surgical instruments, electrode apparatus, and monitoring equipment required for eyelid implantation surgery.

▶ 02:56
3
Prepare animal for eyelid surgery

Anesthetize rat and position for surgery. Verify proper anesthetic depth and maintain physiological monitoring throughout procedure.

▶ 03:13
4
Implant eyelid electrodes surgically

Make incisions around the eye, insert recording and stimulating electrodes into eyelid musculature, and secure apparatus to skull. Allow 7-10 days post-surgical recovery.

▶ 03:50
5
Acclimate animal to conditioning apparatus

Place recovered rat in testing chamber and allow habituation to restraint, electrode connections, and environmental stimuli before formal conditioning begins.

▶ 09:37
6
Verify electrode function and animal status

Confirm proper electrode impedance, baseline eyelid position, and general health status before initiating conditioning trials.

▶ 10:40
7
Execute trace eyeblink conditioning protocol

Present paired conditioned stimulus (tone) and unconditioned stimulus (eyelid shock) with trace interval between them. Record eyelid responses across multiple trials and sessions to assess learning.

▶ 11:08
8
Configure acquisition software and initiate recording

Set trial parameters, stimulus timing, and data collection settings in experimental software. Begin automated data acquisition and electrode signal monitoring.

▶ 15:00
9
Monitor live eyelid response acquisition

Observe real-time eyelid position traces and conditioned response (CR) generation during individual trials to ensure protocol fidelity.

▶ 15:44
10
Analyze adaptive CR development across sessions

Compare conditioned response topography and acquisition curves between control and alcohol-exposed animals across six training sessions to identify hippocampal dysfunction.

▶ 16:46
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