Orthodromic synaptic activation of rat olfactory bulb mitral cells in isolated slices

Axons of olfactory receptor neurons terminate in the glomerular layer of the olfactory bulb, where they synapse with the apical dendrites of mitral cells. Although the mitral cell and its excitation by the olfactory nerve have been the subject of numerous experimental investigations, in vitro studies of these neurons have primarily used nonmammalian preparations. We have recorded the responses of rat olfactory bulb mitral cells to stimulation of the olfactory nerve layer in vitro using extracellular and whole cell patch techniques. Olfactory bulbs were cut into 400-μm thick slices in approximately horizontal section and submerged in a recording chamber. Patch clamp electrodes were guided into the mitral cell layer, which was visible under a dissecting microscope. A stimulating electrode was placed onto the olfactory nerve layer (ONL) rostral to the recording electrode. In extracellular recordings, mitral cells typically responded to ONL stimulation with a prolonged excitation lasting 1 s or longer. With whole cell patch recordings, membrane resistances (mean 272 MΩ) were substantially higher than those reported in previous intracellular studies that used sharp electrodes. Small spontaneous excitatory potentials were present in some mitral cells. ONL stimulation caused a prolonged depolarization comparable to the duration of the period of excitation observed in extracellular recordings. At membrane potentials near −55 mV, ONL stimulation evoked a train of spikes. All but the first of these spikes were blocked by hyperpolarization of the membrane to −65 mV.