Presynaptic localization of Kv1.4-containing A-type potassium channels near excitatory synapses in the hippocampus

Mammalian Shaker voltage-gated potassium channels that contain the Kv1.4 subunit exhibit rapid activation and prominent inactivation processes, which enable these channels to integrate brief (approximiately milliseconds) depolarizations over time intervals of up to tens of seconds. In the hippocampus, Kv1.4 immunoreactivity is detected at greatest density in two regions: (1) the middle molecular layer (MML), where perforant path axons synapse with dentate granule cells, and (2) the stratum lucidum (SL) of CA3, where the mossy fibers travel in tight fasciculi and form en passante synapses onto CA3 pyramidal cells. We have studied the localization of Kv1.4 within these regions in detail. First, we compared the distribution of Kv1.4 and synaptophysin (a synaptic vesicle protein primarily localized near termini) under confocal immunofluorescence microscopy. In the MML, Kv1.4 and synaptophysin immunofluorescence appeared to overlap. In the SL, however, Kv1.4 and synaptophysin staining was detected in nonoverlapping, irregular patches ( approximately 5-10 micro m in diameter). Ultrastructural studies of these two regions revealed that Kv1.4 immunoreactivity was absent from the surface membranes of cell bodies and dendrites and occurred prominently on axons, including axonal "necks" near termini. Small excitatory synaptic boutons also were labeled in the MML; by contrast, the mossy fiber synaptic expansions in the SL were not stained. These localizations may enable Kv1.4-containing channels to regulate the process of neurotransmitter release at these excitatory synapses.