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Frogs (Rana ridibunda) of either sex and of the same age, weighing between 75 g to 110 g, were used. Frogs were bought from local suppliers and kept in tanks at a temper- ature of 18 ± 2 C in 12 h light and 12 h darkness. The animals were anaesthetised, decapitated and pithed. The sciatic nerves of the frog were dissected, from the spinal cord to the knee, and mounted across a three-chambered recording bath made of Plexiglas. The recording system has been described elsewhere (Theophilidis and Pavlidou, 1993; Theophilidis et al., 1997; Kouri and Theophilidis, 2002; Papaefthimiou et al., 2004), but a short description will be given below. The three-chamber recording bath consists of: (a) the recording chamber, where the active electrode of an AC amplifier and the distal part of the nerve, approximately 1 cm in length, were immersed, (b) the perfusion chamber where 80% of the 10–12 cm long nerve and the grounding wires of the amplifier and the stimulator were immersed and (c) the

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