Home › Immunology › Rat Model of Widespread Cerebral Cortical Demyelination Induced by an Intracerebral Injection of Pro-Inflammatory Cytokines
Immunology JoVE (Open Access) Citable · DOI

Rat Model of Widespread Cerebral Cortical Demyelination Induced by an Intracerebral Injection of Pro-Inflammatory Cytokines

DOI: 10.3791/57879-v
What you'll learn
  • ✓ Perform intracerebral catheter implantation in adult rats
  • ✓ Execute subclinical immunization against myelin oligodendrocyte glycoprotein
  • ✓ Inject pro-inflammatory cytokine mixture to induce cortical demyelination
  • ✓ Assess demyelination via PLP immunoreactivity in brain sections
Protocol

The protocol presented here allows the reproduction of a widespread grey matter demyelination of both cortical hemispheres in adult male Dark Agouti rats. The method comprises of intracerebral implantation of a catheter, subclinical immunization against myelin oligodendrocyte glycoprotein, and intracerebral injection of a pro-inflammatory cytokine mixture through the implanted catheter.

Difficulty
advanced
Total time
~3-4 weeks (immunization phase 2-3 weeks; surgical procedure ~2-3 hours; injection ~1 hour; tissue analysis ~1-2 days)
Model organism
Rat Dark Agouti, adult male
Biosafety
BSL-2

Steps

1
Understand demyelination induction methodology

Review the rationale for inducing widespread grey matter demyelination via intracerebral catheterization and pro-inflammatory cytokine injection in the Dark Agouti rat model.

▶ 00:05
2
Implant intracerebral catheter in rat brain

Surgically place a catheter into the cerebral cortex of anesthetized adult male rats using stereotaxic coordinates and aseptic technique.

▶ 01:15
3
Administer subclinical immunization against myelin antigen

Perform immunization of rats against myelin oligodendrocyte glycoprotein to prime the immune response prior to cytokine injection.

▶ 04:39
4
Inject pro-inflammatory cytokines intracerebrally

Deliver pro-inflammatory cytokine mixture through the implanted catheter to induce widespread cortical demyelination in both hemispheres.

▶ 05:57
5
Analyze PLP immunoreactivity in brain sections

Prepare coronal brain sections and perform immunohistochemistry for proteolipid protein to quantify demyelination extent across cortical regions.

▶ 07:16
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