Locke's solution was used for extracellular recording, of which composition was: 140 mM NaCl, 5.6 mM KCl, 1.2 mM Mg[Cl.sub.2], 2.2 mM Ca[Cl.sub.2], 10 mM tris(hydroxymethyl- aminomethane), and 10 mM glucose, pH adjusted to 7.4, with HCl.
The nerve was immediately placed in a Petri dish containing ice cold (4[degrees]C) modified Locke's solution and they were used for experimental recording in the same day.
The preparations were placed in isolated organ baths, incubated in Locke's solution at 37 [+ or -] 1[degrees]C and bubbled with gas (95% [O.sub.2], 5% C[O.sub.2]).
A uterine horn was incubated in Locke's solution and equilibrated for 45 min.
As is well known, Locke's solution in the Letter opposes the state enforcing religious practice.
Ironically, Locke's solution recreates one of the key problems of state-controlled religion: the nature of the church's relation to the state prevents one from addressing the other.
For nerve dissection and extracellular recordings, modified Locke's solution was used with the following composition: 140 mM NaCl, 5.6 mM KCl, 2.2 mM Ca[Cl.sub.2], 1.2 mM Mg[Cl.sub.2], 10 mM glucose, 10 mM Tris-hydroxymethyl aminomethane (TRIS), pH adjusted to 7.4 with HCl or NaOH.
The sciatic nerves were stored in modified Locke's solution at room temperature until electrophysiological recordings.
A 15-20-mm segment of the nerve suspended between the stimulating and recording electrodes was immersed in Locke's solution, which was employed to maintain chamber humidity.
Modified Locke's solution (composition in mM: 140 NaCl, 5.6 KCl, 2.2 Ca[Cl.sub.2], 1.2 Mg[Cl.sub.2], 10 Glucose, and 10 TRIS-(Hydroxymethyl)Aminomethane) (pH 7.4) was thoroughly aerated before use in the chamber.
After rats were killed by C[O.sub.2] inhalation, DRG were dissected and immediately placed in an ice-cold modified Locke's solution
. For intracellular recordings, intact tissues were used on the same day of dissection.
Each testing extract was dissolved in Locke's solution
(in mM: 154 NaCl, 5.6 KCl, 1.2 Mg, 3.6 NaHC[O.sub.3], 5.0 HEPES, 2.3 Ca[Cl.sub.2], 5.6 glucose, pH 7.4) as X 100 concentrated stock solution, and added (20 [micro]l) to the original culture medium.