Inhibitory effects of glutamate-stimulated astrocytes on action potentials evoked by bradykinin in cultured dorsal root ganglion neurons.

Patch-clamp and Ca2+-imaging techniques have revealed that astrocytes have dynamic properties including ion channel activity and release of neurotransmitters, such as adenosine triphosphate (ATP) and glutamate. Here, we used the patch-clamp technique to determine whether ATP and glutamate is able to modulate the bradykinin (BK) response in neurons cultured with astrocytes in the mouse dorsal root ganglia (DRG) in order to clarify the role of astrocytes in nociceptive signal transmission. Astrocytes were identified using a fluorescent anti-GFAP antibody. The membrane potential of astrocytes was about -39 mV. The application of glutamic acid (GA) to the bath evoked the opening of two types of Cl- channel in the astrocyte cell membrane with a unit conductance of about 380 pS and 35 pS in the cell-attached mode, respectively. ATP application evoked the opening of two types of astrocyte K+ channel with a unit conductance of about 60 pS and 29 pS, respectively. Application of BK to the neuron evoked an action potential (spike). Concomitant BK application with ATP increased the frequency of BK-evoked neuron spikes when neurons coexisted with astrocytes. Stimulation of BK with GA inhibited the BK-evoked spike under similar conditions. The application of furosemide, a potent cotransporter (Na+-K+-2Cl-) inhibitor, prior to stimulation of BK with GA blocked inhibition of the spike. It is thought that inhibition of the spike is related to Cl- movement from astrocytes.