The chloride homeostasis of neurons and non-neuronal cells is maintained in part by a voltage-sensitive inwardly rectifying chloride conductance through the chloride channel-2. This channel is activated by hyperpolarization and extracellular hypotonicity. In the present study, hippocampal sections were immunostained for chloride channel-2, and somata and dendrites of both pyramidal and non-pyramidal cells were found to be immunoreactive. In addition, glial processes in the vicinity of small blood vessels were also immunostained, whereas the neuropil of strata pyramidale and lacunosum-moleculare contained chloride channel-2-positive punctate structures. Electron microscopy and double immunostaining using antibodies against chloride channel-2 and glial fibrillary acidic protein confirmed that the dense network of chloride channel-2-positive processes corresponds to the end feet of astrocytes. The distribution of chloride channel-2-immunoreactive astrocytes was inhomogeneous throughout the hippocampus: strata oriens, pyramidale and lacunosum-moleculare of CA1-CA3 and the outer molecular layer of the dentate gyrus contained the majority of immunoreactive end feet, whereas the other layers showed sparse labeling. Subcellular studies demonstrated that, in addition to astrocytes, chloride channel-2 was localized in the membrane of dendrites, dendritic spines, cell bodies and axon initial segments of neurons, frequently close to, or within active zones of, symmetrical synapses.Thus, chloride channel-2 appears to be involved in transmembrane chloride movements associated with GABAergic synaptic transmission. The specific laminar distribution of chloride channel-2-positive astroglial processes coinciding with that of GABAergic axon terminals suggests that the network of astrocytes may be able to siphon and deliver Cl(-) ions to layers with intense GABAergic transmission, thereby increasing the efficacy of GABA(A) receptor-mediated inhibition.