Intracellular Cl- activity (aCli) and 5-hydroxytryptamine (5-HT)-induced membrane currents of Retzius neurons in the central nervous system of the medicinal leech were measured using Cl- sensitive microelectrodes and a two-microelectrode voltage-clamp technique. At the membrane of Retzius neurons Cl- ions were not passively distributed. Under different conditions the chloride equilibrium potential (ECl, -60.1 mV for isotonic saline and -57.8 mV for a hypertonic saline) was negative with respect to the membrane potential (Em, -55 +/- 3.8 and -47 +/- 3.4 mV respectively). The endogenous neurohormone 5-HT always polarized the membrane of Retzius neurons in the direction of ECl. When voltage-clamping the membrane of Retzius neurons near the resting potential both in situ and in primary culture, application of 5-HT produced an outward current (I5-HT) and an increase in membrane conductance. Current-voltage relationships for I5-HT showed a slight outward rectification and reversal potentials of -61.6 +/- 3.1 mV in situ and -66 +/- 3.1 mV in primary culture, both values being comparable to the ECl of Retzius neurons as measured in situ. The results indicate that 5-HT increases the Cl- conductance of Retzius neurons, thereby hyperpolarizing the cell membrane and affecting both the excitability of the neuron and 5-HT release from it. This could affect the feeding and swimming behaviour of the leech.