Functional and structural correlates of cell size in paracerebral neurons of Pleurobranchaea californica.

1. The paracerebral neurons (PCNs) in the brain of the mollusk Pleurobranchaea are a population of 12-16 interneurons that send axons to the buccal ganglion and control cyclic feeding behavior (9). In the present study we show that the PCNs differ in size and that a number of functional and structural properties of the PCNs are closely correlated with cell size. 2. PCN soma diameter varies from about 30 to 120 micrometers. The diameters segregate into two distinct but overlapping populations, which correspond to independently assigned functional classifications of "tonic" and "phasic" PCNs. The mean soma diameters of two populations were 63 and 84 micrometers, respectively. 3. Two morphological features vary systematically with PCN soma size. First, soma diameter, axonal conduction velocity, and extracellular spike amplitude were positively correlated; therefore, PCN axon diameter presumably increases with soma diameter. Second, intrasomatic injection of lucifer yellow revealed that the small, tonic PCNs are multipolar, while the large, phasic PCNs are generally monopolar neurons. 4. Small PCNs discharge tonically in response to sustained current injection and have a weak effect on cyclic motor output recorded from nerves that innervate feeding muscles. In contrast, the large PCNs discharge phasically in bursts of action potentials that are coordinated with the cyclic motor output and have a comparatively strong effect on the rhythm. The motor effects of simultaneous tonic and phasic PCN stimulation are additive. 5. Tonic and phasic PCNs innervate different but partially overlapping populations of feeding motor neurons. Phasic PCNs typically inhibit motor neurons exiting buccal root 3, while tonic PCNs either have no effect or are weakly excitatory. 6. Tonic and phasic PCNs exhibit different intrinsic properties. In comparison with phasic PCNs, tonic PCNs have higher input resistances, higher spontaneous discharge rates at rest potential, lower firing thresholds to intrasomatically injected current, lower absolute voltage thresholds, greater pacemaker sensitivity, and greater total capacitance. 7. Tonic and phasic PCNs exhibit different input properties. Tonic PCNs are recruited before phasic ones during cycylic buccal motor output induced by stomatogastric nerve stimulation. Phasic PCNs receive powerful, cycylic inhibition that is not shared by tonic PCNs. In addition, extracellular stimulation of the large oral veil nerve of the brain excites tonic PCNs but causes a biphasic postsynaptic potential (PSP) in phasic PCNs that has a net inhibitory effect. Some excitatory synaptic input to phasic and tonic PCNs is unshared, while some is shared. 8. It is concluded that these command interneurons obey the size principle discovered earlier in motor neurons (4, 13-16). Cell size per se is not the causal variable, however; instead the underlying causes of the differences between small and large PCNs include different input and output organizations as well as different intrinsic functional and morphological properties.