Inducing Hypoxic-Ischemic and Inflammatory Conditions to Model Encephalopathy in Prenatal Rats

Take an anesthetized pregnant rat with exteriorized uterine horns containing fetuses positioned on moist gauze.

Place aneurysm clips on the exposed uterine arteries to restrict blood flow, evidenced by darkening uterine vessels.

This restriction induces hypoxic-ischemic conditions by depriving the fetuses of oxygen and nutrients, leading to central nervous system damage.

Regularly hydrate the uterine horns with sterile saline.

After the occlusion period, rinse the uterine horns and remove the aneurysm clips to restore blood flow.

Inject a lipopolysaccharide or LPS and dye mixture into the amniotic fluid of each amniotic sac, using the dye to confirm the injection visually.

Rinse the uterine horns, return them to the peritoneal cavity, and close the incision.

In the amniotic fluid, LPS binds to immune cell receptors, triggering the release of pro-inflammatory cytokines.

These cytokines reach the fetal brain, activate microglial cells, and cause neuroinflammation, resulting in brain damage and fetal encephalopathy.

When the arteries have been isolated, place a 30 gauge rat aneurysm clip on each vessel.

Before applying the clips, it's important to ensure that all the surrounding tissue is co