The effect of glutamatergic agonists on the intracellular free Ca2+ concentration ([Ca2+]i) of neuropile glial cells and Retzius neurones in intact segmental ganglia of the medicinal leech Hirudo medicinalis was investigated by using iontophoretically injected fura-2. In physiological Ringer solution the [Ca2+]i levels of both cell types were almost the same (glial cells: 58 +/- 30 nM, n = 51; Retzius neurones: 61 +/- 27 nM, n = 64). In both cell types glutamate, kainate, and quisqualate induced an increase in [Ca2+]i which was inhibited by 6,7-dinitroquinoxaline-2,3-dione (DNQX). This increase was caused by a Ca2+ influx from the extracellular space because the response was greatly diminished upon removal of extracellular Ca2+. The glutamate receptors of neuropile glial cells and Retzius neurones differed with respect to the relative effectiveness of the agonists used, as well as with regard to the inhibitory strength of DNQX. In Retzius neurones the agonist-induced [Ca2+]i increase was abolished after replacing extracellular Na+ by organic cations or by mM amounts of Ni2+, whereas in glial cells the [Ca2+]i increase was largely preserved under both conditions. It is concluded that in Retzius neurones the Ca2+ influx is predominantly mediated by voltage-dependent Ca2+ channels, whereas in neuropile glial cells the major influx occurs via the ion channels that are associated with the glutamate receptors.