Biology Reference
In-Depth Information
24. Lubricant eye ointment (artificial tears, 15% mineral oil in 83%
white petrolatum).
25. Heparin to withdraw blood (2 IU/ml of heparin in 0.9%
saline).
26. Citrate phosphate dextrose anticoagulant (to be used at a ratio
to whole blood = 1.4:10).
1. Beam walking: wooden beam (1 m long and 2.5 cm wide) for
rats, plastic wire (50 cm long and 2 mm thick) for mice, light
source, darkened goal box, noise (90 db) generator, timer, and
four plastic pegs (7.5 mm long and 2.5 mm wide).
2. Beam balancing: wooden beam (1 m long and 1 cm wide for
rats) and a wire of 2 mm thick for mice, large plastic container
(120 cm × 40 cm and 60 cm high).
3. Morris water maze: galvanized steel pool (5 foot diameter),
plexiform platform (10 × 10 cm), water, and nontoxic, white
paint, heating lamp, and Chromotrack Video tracking system
(Sam Diego Instruments, San Diego, CA). Computer software
HVS image.
2.3 Behavioral
Studies
2.4 Tissue
Processing
1. 4% paraformaldehyde solution; ddH 2 O, 1 N NaOH, pH strip,
paraformaldehyde (Sigma), sodium phosphate monobasic/
monohydrate, sodium phosphate dibasic anhydrous, and hot
stir plate.
2. Chemical fume hood
3. Personal protective equipment: safety glasses, sterile apron,
cap, and mask.
4. 30% sucrose.
3
Methods
3.1
Anesthesia
1. Rats: Anesthesia is induced by placing rats in the anesthesia
chamber with 5% isoflurane in 100% oxygen flow for 3-5 min.
Then the rats are intubated with ET tubes made from 14 G/16
G catheters using an intubation stand (a plexiglass stand that
allows suspending the rat by its incisors, on its sloping side
using an elastic band) and a laryngoscope (infant blade). Using
the laryngoscope, the vocal cords are visualized and catheter is
placed into the trachea. The ET tubes are prepared using
14/16 G catheter, its stylet and the cut proximal portion of
No. 2.5 (ID) ET tube as shown in Fig. 2a . The ET tube is
secured by tying it to the lower jaw with an elastic tape.
Anesthesia is maintained with 2% isoflurane in 100% oxygen
(see Note 1).
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