Leech blood apparently contains considerably less chloride than generally used in physiological experiments. Instead of 85-130 mM Cl- used in experimental salines, leech blood contains around 40 mM Cl- and up to 45 mM organic anions, in particular malate. We have reinvestigated the distribution of Cl- across the cell membrane of identified glial cells and neurones in the central nervous system of the leech Hirudo medicinalis L., using double-barrelled Cl(-)- and pH-selective microelectrodes, in a conventional leech saline, and in a saline with a low Cl- concentration (40 mM), containing 40 mM malate. The interference of anions other than Cl- to the response of the ion-selective microelectrodes was estimated in Cl(-)-free salines (Cl- replaced by malate and/or gluconate). The results show that the absolute intracellular Cl- activities (aCli) in glial cells and neurones, but not the electrochemical gradients of Cl- across the glial and the neuronal cell membranes, are altered in the low Cl-, malate-based saline. In Retzius neurones, aCli is lower than expected from electrochemical equilibrium, while in pressure neurones and in neuropil glial cells, aCli is distributed close to its equilibrium in both salines, respectively. The steady-state intracellular pH values in the glial cells and Retzius neurones are little affected (< or = 0.1 pH units) in the low Cl-, malate-based saline.