A whole-cell clamp study of dendrodendritic synaptic activities in mitral cells of turtle olfactory bulb slices

A procedure for preparing slices from the turtle olfactory bulb is described in this report. Individual layers of the bulb could be identified in the slices which enabled visual identification of cell types. Mitral cells retained extensive dendritic arborizations in slices of typical thickness, 300-400 microm. The presence of extensive dendritic processes was consistent with the difficulties we encountered in our attempt to achieve adequate space clamp. On the few occasions where an adequate space clamp of a mitral cell was achieved, calcium current exhibited a threshold of - 50 mV and reached its maximal level at - 10 mV. In all cases where calcium current was analysed (n=46), the current exhibited little inactivation. Depolarizing steps in 50% of the mitral cells triggered a burst of feedback synaptic activity after termination of the step. The intensity of feedback activity correlated closely with the amplitude of the depolarizing step, reaching its maximal level at - 10 mV and declining with further depolarization. The bell-shaped relationship between the feedback activity and mitral cell depolarization is consistent with the hypothesis that the feedback activity is mediated by reciprocal synapses on the mitral cell dendrite. This hypothesis is further supported by the inhibitory nature of the feedback synaptic activity: (i) the polarity of the feedback synaptic current could be inverted at the predicted chloride equilibrium potential, (ii) the feedback activity could be completely blocked by 10 microM bicuculline. The analysis of spontaneous synaptic activity showed that it was mostly inhibitory because its polarity could be reversed at the predicted chloride equilibrium potential. In some mitral cells, the frequency of spontaneous activity was noticeably increased when the holding potential was depolarized. This correlation could be attributed to the activation of dendrodendritic synapses. Results shown in this report demonstrate that dendrodendritic synapses are viable in turtle olfactory bulb slices. In addition, the suppression of feedback inhibition by large depolarizing steps of mitral cells suggests that the control of mitral cell dendritic potential is adequate to suppress calcium influx during large depolarizing steps.