Erythropoietin as a Neuroprotectant for Neonatal Brain Injury: Animal Models - Tissue-Protective Cytokines: Methods and Protocols - page 113

1.4 Overview
of Procedure

Neonatal rat pups are anesthetized, undergo isolation and electro-
cauterization of the right common carotid artery, followed by
exposure to 8% oxygen for 90 min to produce a moderate unilat-
eral hypoxic-ischemic brain injury. Depending on the length of the
hypoxic episode, the timing of peak cerebral edema ranges from 4
to 24 h ( 23, 59 ). The blood-brain barrier is altered during the
recovery from the hypoxic injury as evidenced by extravasation of
horseradish peroxidase from the vascular space into the brain
parenchyma ( 60 ). Cerebral blood flow to the ipsilateral brain is
unaffected by the unilateral common carotid ligation, but is
decreased during the hypoxia-ischemic episode ( 12 ) (see Note 1).

2 Materials

1. Personal protective equipment (lab coat, sterile gloves, mask).
2. Instrument sterilizer.
3. Inhalant anesthetic (e.g., Isoflurane), matching vaporizer with
regulated oxygen supply and adjustable flow control.
4. Anesthesia induction chamber.
5. Anesthetic exhaust or trap.
6. Dissecting microscope with light source.
7. Plastic animal cradle. Should contain a V-shaped indentation in
which to hold the rat pup supine during surgery (Fig. 1 ).
8. Heating pad × 2.
9. Small animal incubator (Veterinary warmer).
10. Fine rat-tooth forceps.
11. Scalpel with #11 blade.
12. Microretractor or suitable substitute.
13. Sharp fine forceps × 2.
14. Cauterizer.
15. Skin staples, skin glue, or suture material for wound closure.
16. Gauze or cotton swabs.
17. Pentobarbital (or other approved euthanizing agent, to eutha-
nize failed surgeries).
18. Body bag.
19. Hypoxia chamber (sealed container with ports for gas
delivery).
20. Thermoregulated water bath.
21. Thermometer.
22. 8% oxygen tank with regulator.
23. Oxygen meter.