Biology Reference
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8. It is important to record the baseline values (blood pressure, arte-
rial blood gases, and intracranial pressures) before the impact.
9. Animals that do not awake within 1 h after the impact are
euthanized. It is important to empty the rebreathing bag after
skin closure and to stop the isoflurane as the gas in rebreathing
bag can delay the recovery from anesthesia.
10. If performing surgeries in batches it is advisable to sterilize the
equipment between two procedures using a hot glass bead
sterilizer.
11. Behavior tests should be conducted in a separate room with no
visual/noise disturbances. During the days for which the
behavior tests are scheduled, the drugs (study drug, analgesic,
and antibiotic) are given after the trials to reduce the stress on
the rats which can affect the behavior tests. If the rats are above
350 g to start with and chronic studies of longer duration are
planned, they are likely to grow further (>400 g) and find it
difficult to perform well in the behavior tests.
12. The pegs in the beam walking test are not bent too much oth-
erwise the task will be too easy for the rats. There should be
soft bedding on the floor below the beam walking and beam
balancing rods, to prevent injury if the rat falls.
13. During the trials it is better for the researcher to stand in a
fixed position so as to serve as a fixed visual clue for the rats.
Do not keep the heating lamp too close to the rat. Clean up
the water maze tank once in a week and follow the sanitizing
methods recommended by the institutional protocols.
14. The bone around the craniectomy defect is removed with gen-
tle force with outwards movement of the rongeurs. Magnifying
loops can be used to avoid damage to the normal brain.
15. Titration is done using 1 N NaOH as the pH meter will be
damaged by paraformaldehyde. Paraformaldehyde powder and
solutions should be handled in the hood and after donning the
personal protective equipment.
16. Prepared solutions (Buffer, 4% paraformaldehyde, 30% sucrose)
are stored at 4°C.
References
1. Langlois JA, Rutland-Brown W, Wald MM
(2006) The epidemiology and impact of trau-
matic brain injury: a brief overview. J Head
Trauma Rehabil 21:375-378
2. McAllister TW et al (2001) Differential work-
ing memory load effects after mild traumatic
brain injury. Neuroimage 14:1004-1012
3. Cherian L, Goodman CJ, Robertson CS
(2007) Neuroprotection with erythropoietin
administration following controlled cortical
impact injury in rats. J Pharmacol Exp Ther
322:789-794
4. Cherian L, Goodman JC, Robertson C (2011)
Improved cerebrovascular function and
reduced histological damage with darbepoietin
alfa administration after cortical impact injury
in rats. J Pharmacol Exp Ther 337:451-456
5. Velly L et al (2010) Erythropoietin 2nd cere-
bral protection after acute injuries: a double-
edged sword? Pharmacol Ther 128:445-459
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