Distribution and functional properties of glutamate receptors in the leech central nervous system

1. The effect of kainate and other glutamatergic agonists on the membrane potential (E(m)), the intracellular Na+ activity (aNa(i)), and the intracellular free Ca2+ concentration ([Ca2+](i)) of identified leech neurons and neuropile glial cells was measured with conventional and ion-sensitive microelectrodes, as well as with the use of the iontophoretically injected fluorescent indicators sodium-binding benzofuran isophthalate and Fura-2. 2. In Retzius neurons, AE, L, 8, and 101 motoneurons, and in the unclassified 50 neurons (Leydig cells) and AP neurons, as well as in neuropile glial cells, bath application of 100 mu M kainate evoked a marked membrane depolarization and an increase in aNa(i) and [Ca2+](i). The kainate-induced aNa(i) increase persisted in solutions with high Mg2+ concentration in which synaptic transmission is blocked. 3. A membrane depolarization as well as an Increase in aNa(i) and [Ca2+](i) was also evoked by L-glutamate. quisqualate, and L-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA). The agonist-induced [Ca2+](i) increase was inhibited by 6,7-dinitroquinoxaline-2,3-dione (DNQX). 4. In Ca2+-free solution, the kainate-induced [Ca2+](i) increase was abolished in the neurons and in neuropile glial cells, whereas membrane depolarization and aNa(i) increase were unchanged, In Na+-free solution, kainate had no effect on E(m), aNa(i), or [Ca2+](i) in the neurons. 5. In the mechanosensory T, P, and N neurons, kainate induced considerably smaller membrane depolarizations than in the other neurons or in neuropile glial cells, and it had no significant effect on aNa, or [Ca2+](i). 6. It is concluded that in leech segmental ganglia the majority of the neurons and the neuropile glial cells, but probably not th; mechanosensory neurons, possess glutamate receptors of the AMPA-kainate type. In the neurons, the [Ca2+](i) increase caused by glutamatergic agonists is due to Ca2+ influx through voltage-dependent Ca2+ channels that are activated by the agonist-induced membrane depolarization.