Electrophysiological Evaluation of Synaptic Plasticity in Injured CNS - Animal Models of Acute Neurological Injuries: Injury and Mechanistic Assessments - page 120

8. A gravity-fed perfusion system.
9. Computer with software for data acquisition and analysis
(e.g., pClamp from Molecular Devices).

1. Cutting solution contains the following three groups of ingre-
dients (in mM):
(a) 252 sucrose, 10 glucose.
(b) 3 KCl, 0.2 CaCl 2 , 6 MgSO 4 , 1.3 NaH 2 PO 4 .
(c) 26 NaHCO 3 .
2. Artifi cial cerebrospinal fl uid (ACSF) contains the following
two groups of ingredients (in mM):
(d) 124 NaCl, 2 KCl, 2 CaCl 2 , 1 MgSO 4 , 1.3 NaH 2 PO 4 , 10
dextrose.
(e) 26 NaHCO 3 .
For storage, stock solutions (a-e) at 10× concentration should
be individually prepared and stored in refrigerator at 4°C. Final
solution is made by mixing together equal amount of each solution
(a-c solutions for cutting solution, d and e solutions for ACSF),
and diluted to fi nal concentration before use. The cutting solution
is then ice-colded, and is bubbled with 95% O 2 and 5% CO 2 .

2.3. Solutions

1. A bipolar tungsten electrode is used as a stimulating electrode
(Fig. 3b , WPI).
2. Recording electrodes are made from borosilicate glass capillaries
(Fig. 3a , b, WPI, 1B150F-4) and have a resistance of 1-3 M

2.4. Stimulating and
Recording Electrodes

Ω

when fi lled with ACSF solution.

3. Procedures

1. A rat is anesthetized with 50 mg/kg pentobarbital sodium i.p.
2. After decapitation, the brain is rapidly removed with a spatula
and submerged in ice-cold cutting solution for 1 min.
3. Place the brain on a small piece of fi lter paper with ventral side
on bottom; make two parallel coronal cuts with one slightly
anterior to the cerebellum and the other at about one-third of
cortex length from the olfactory bulb (Fig. 2a ).
4. The brain is glued onto the cutting stage of a Vibratome with
instant glue, with the caudal surface on the bottom and dorsal
surface facing the blade (Fig. 2b ). A cut along the midline may
be made carefully so that the two hemispheres can be separated.
A block of 4% agar should be glued on the ventral side of the
brain, which provides support to the brain on the opposite side
against the direction of blade movement (Fig. 2b ).

3.1. Preparation of
Cortical and
Hippocampal Slices

3.1.1. Neocortical Slices'
Preparation