1. As part of a continuing investigation of the organization of the spinal cord of the lamprey, propriospinal interneurons with axons projecting contralaterally and caudally (CC interneurons) were surveyed with intracellular recordings. 2. CC interneurons were identified by recording their axon spikes extracellularly in the spinal cord during intracellular stimulation of the cell body. The axon projections of Cc interneurons were confirmed after intracellular injection and development of horseradish peroxidase. 3. Intracellular stimulation of CC interneurons produced synaptic potentials in myotomal motoneurons, lateral interneurons and other CC interneurons that lay caudally on the opposite side of the spinal cord. Most CC interneurons were inhibitory, but some were excitatory. 4. CC interneurons were divided into three classes on the basis of reticulospinal Müller cell inputs. CC1 interneurons were excited by the ipsilateral Müller cell B1 and the contralateral Mauthner cell. CC1 interneurons were inhibitory. They were excited polysynaptically by ipsilateral sensory dorsal cells and were inhibited by contralateral dorsal cells. They were distinguished morphologically by having no rostral axon branch and no contralateral dendrites. CC1 interneurons were phasically active during fictive swimming with their peak depolarizations preceding those of myotomal motoneurons by about 0.15 cycle. 5. CC2 interneurons were also inhibitory, but they were distinguished from CC1 interneurons by their excitation from the ipsilateral Müller cells B2-4 nd by their thin rostral and thicker caudal axonal branches on the contralateral side of the spinal cord. 6. CC3 interneurons were excitatory, and they were inhibited by the ipsilateral Müller cell I1. CC3 interneurons could have contralateral dendrites and bifurcating axons, and they had lower average axonal conduction velocities than CC1 and CC2 interneurons. 7. Inhibitory CC interneurons may be important for motor coordination in the lamprey. Movements of the lamprey body during reflexes and swimming consist of contraction and relaxation of myotomal muscles on opposite sides of the body. By being coactive with ipsilateral myotomal motoneurons, inhibitory CC interneurons could contribute to the inhibition of contralateral motoneurons during these movements.

A new class of excitatory premotor interneurons that are important in the generation of locomotion in the lamprey has now been described. In the isolated spinal cord, these neurons act simultaneously with their postsynaptic motoneurons during fictive swimming. They are small and numerous, and they monosynaptically excite both motoneurons and inhibitory premotor interneurons. The excitatory postsynaptic potentials are depressed by an antagonist of excitatory amino acids. These interneurons receive reticulospinal input from the brain stem and polysynaptic input form skin afferents. A model of the network underlying locomotion based on the synaptic interactions of these neurons can now be proposed for the lamprey.

Among the advantages offered by the lamprey brainstem and spinal cord for studies of the structure and function of the nervous system is the unique identifiability of several pairs of reticulospinal neurons in the brainstem. These neurons have been exploited in investigations of the patterns of sensory input to these cells and the patterns of their outputs to spinal neurons, but no doubt these cells could be used much more effectively in exploring their roles in descending control of the spinal cord. The variability of cell positions of neurons in the spinal cord has precluded the recognition of unique spinal neurons. However, classes of nerve cells can be readily defined and characterized within the lamprey spinal cord and this has led to progress in understanding the cellular and synaptic mechanisms of locomotor activity. In addition, both the identifiable reticulospinal cells and the various spinal nerve cell classes and their known synaptic interactions have been used to demonstrate the degree and specificity of regeneration within the lamprey nervous system. The lack of uniquely identifiable cells within the lamprey spinal cord has hampered progress in these areas, especially in gaining a full understanding of the locomotor network and how neuromodulation of the network is accomplished.

1. Application of D-glutamate to the isolated spinal cord of the lamprey produces phasic activity in ventral roots, which is similar to that of the muscles of the intact swimming animal (5,18). Therefore, the isolated spinal cord may be used as a convenient model for the investigation of the generation of locomotor rhythms in a vertebrate. 2. Almost all slow muscle fibers exhibited excitatory junctional potentials (EJPs) during swimming activity. The number of EJPs per cycle increased with the intensity of ventral root (VR) bursting. Few twitch fibers were active, and these fired action potentials only during high intensities of VR bursts. 3. As was found by Russell and Wallén (25), myotomal motoneurons had oscillating membrane potentials during fictive swimming which, on the average, reached a peak depolarization in the middle of the VR burst (phi = 0.21 +/- 0.05; phi = 0 is defined as the onset of the VR burst, and the duration of the cycle is set equal to 1). Membrane potential oscillations in fin motoneurons were antiphasic to those of nearby myotomal motoneurons (peak depolarization phi = 0.68 +/- 0.05). 4. Lateral interneurons had oscillating membrane potentials in synchrony with those of myotomal motoneurons (peak depolarization phi = 0.21 +/- 0.10). Interneurons with axons projecting contralaterally and caudally (CC interneurons) had oscillating membrane potentials that peaked significantly earlier in the cycle (peak depolarization phi = 0.06 +/- 0.12). 5. Edge cells were only weakly modulated during fictive swimming. Their peak depolarizations occurred near the end of the VR burst (phi = 0.33 +/- 0.10). Most giant interneurons were not phasically modulated during fictive swimming. 6. Repetitive intracellular stimulation of Müller cells during fictive swimming generally evoked an increased burst intensity in ipsilateral VRs and a decreased burst intensity in contralateral VRs. The cells M3, B1, and B2 also produced increases or decreases in the frequency of VR bursts. Repetitive intracellular stimulation of sensory dorsal cells could also change the intensities and timing of VR bursts. 7. This study is an initial survey of lamprey spinal interneurons that participate in swimming activity. Lateral interneurons and CC interneurons are active during fictive swimming and probably help coordinate the undulations of the body, but their roles in pattern generation are not known. The central pattern generator is subject to modification by descending and sensory inputs.

1. Local application of 5-hydroxytryptamine (5-HT) in the area in which a dense 5-HT plexus is located in the lamprey spinal cord leads to a marked depression of the late phase of the afterhyperpolarization (AHP) following the action potential. This effect was observed in motoneurons, premotor interneurons, and giant interneurons, whereas no effect was observed in the sensory dorsal cells and edge cells. 2. The late 5-HT sensitive phase of the AHP was increased in amplitude when calcium entry was enhanced during the prolongation of action potentials caused by tetraethylammonium (TEA). Conversely, a blockade of Ca2+ entry by manganese reduced the AHP amplitude, suggesting that a calcium-dependent current, most likely carried by potassium, underlies the late phase of the AHP in these cells, as is the case in many other types of neurons. 3. The late phase of the AHP could be depressed by 5-HT although no effects were exerted on either the resting input resistance or on the shape of the action potential in 54% of the cells. The membrane conductance increase associated with the late phase of the AHP was markedly attenuated by 5-HT application. 4. In voltage-clamp experiments, Na+ currents and most K+ currents were blocked by tetrodotoxin (TTX) and TEA, respectively. Under these conditions, voltage steps elicited a slow outward current, most likely representing a Ca2+-activated K+ current, which was depressed by 5-HT application. 5. 5-HT does not appear to reduce AHP amplitude by blocking the calcium entry occurring during the action potential. No evidence was obtained for an involvement of second messengers such as adenosine-3':5'-cyclic monophosphate (cAMP), guanosine-3':5'-cyclic monophosphate (cGMP), diacyglycerol, or arachidonic acid. The effect of 5-HT on the late AHP may be due to a direct action on the calcium-dependent potassium channels or on the intracellular handling of Ca2+ ions. 6. The amplitude reduction of the AHP has a profound influence on the spike frequency regulation of any given cell; the frequency of spikes evoked by a given excitatory stimulus is therefore markedly increased by application of 5-HT. 5-HT thus increases the "gain" of the input-output relation of interneurons and motoneurons responsible for generating the locomotor rhythm. In addition, 5-HT causes a prolongation of the depolarized plateau of the N-methyl-D-aspartate (NMDA) receptor-induced membrane potential oscillations, as expected from the 5-HT-induced effects on the Ca2+-activated K+ channels that contribute to the repolarization.

1. In the in vitro preparation of the lamprey spinal cord, paired intracellular recordings of membrane potential were used to identify interneurons producing excitatory postsynaptic potentials (EPSPs) on myotomal motoneurons. 2. Seventy-nine interneurons (8.4% of all neuron-motoneuron pairs tested) elicited unitary EPSPs that followed one-for-one at short, constant latencies and were therefore considered monosynaptic according to conventional criteria. Evidence was obtained for selectivity and divergence of excitatory interneuron (EIN) outputs and for convergence of EIN input to motoneurons. 3. The neurotransmitter released by EINs may be an excitatory amino acid such as glutamate, because the EPSPs were depressed by antagonists of excitatory amino acids. 4. Intracellular dye injection revealed that EINs have small cell bodies (average 11 x 27 microns), transversely oriented dendrites, and thin (less than 3 microns) slowly conducting axons (0.7 m/s) that project caudally and ipsilaterally. One EIN exhibited a system of thin multi-branching axon collaterals with periodic swellings. Ultrastructurally, these swellings contained clear spherical vesicles, and they apposed postsynaptic membrane specializations. 5. During fictive locomotion, the membrane-potential oscillations of EINs were greater in amplitude than, but similar in shape and timing to, those of their postsynaptic motoneurons. EINs fired action potentials during fictive locomotion and contributed to the depolarization of motoneurons. 6. These interneurons are proposed to be a source of excitation to motoneurons and interneurons in the lamprey spinal cord, participating in motor activity including locomotion.

Swimming movements in the leech and lamprey are highly analogous, and lack homology. Thus, similarities in mechanisms must arise from convergent evolution rather than from common ancestry. Despite over forty years of parallel investigations into this annelid and primitive vertebrate, a close comparison of the approaches and results of this research is lacking. The present review evaluates the neural mechanisms underlying swimming in these two animals and describes the many similarities that provide intriguing examples of convergent evolution. Specifically, we discuss swim initiation, maintenance and termination, isolated nervous system preparations, neuralcircuitry, central oscillators, intersegmental coupling, phase lags, cycle periods and sensory feedback. Comparative studies between species highlight mechanisms that optimize behavior and allow us a broader understanding of nervous system function.

1. As part of a continuing analysis of the mechanisms of the central pattern generator underlying fictive swimming in lamprey, a systematic survey of electrophysiological properties of lamprey neurons was made in the in vitro spinal cord preparation with the use of intracellular current-clamp recordings. A total of 70 neurons was included in the study, representing 6 classes of spinal neurons. The classes were myotomal motoneurons, three classes of interneurons involved in fictive swimming [lateral interneurons, nerve cells with contralateral and caudal projecting axons (CC interneurons), and excitatory interneurons], and two classes of interneurons involved in sensory processes (edge cells and giant interneurons). The recordings were done in quiescent preparations. 2. There was little or no significant difference among the cell classes with regard to resting potential, threshold potential, action-potential amplitude, or action-potential duration. 3. The voltage versus current relationships for the cells were fairly linear near resting potential, although most cells showed a slight tendency to rectify with depolarization above resting potential. This tendency was strongest among edge cells and lateral interneurons and weakest among motoneurons and CC interneurons. The input resistances, membrane time constants, and rheo-bases for the cell classes showed significant differences among some classes. For example, CC interneurons and excitatory interneurons had significantly higher input resistances than the other cell classes. 4. The late afterhyperpolarization following the action potential tended to be larger in amplitude with an earlier peak and a longer duration in edge cells and giant interneurons than in the other cell classes. 5. All cells responded to depolarizing current injections by firing action potentials, and almost all cells fired action potentials throughout the 400-ms current pulse. The cells exhibited adaptation resulting in increasing interspike intervals during the current pulse. The adaptation, however, was insufficient to terminate firing before the end of the current pulse. The relationship between frequency of firing and input current was generally monotonic with a tendency to saturate at higher current levels. 6. The general conclusion from this study is that the spinal neurons that partake in fictive swimming (motoneurons, lateral interneurons, CC interneurons, and excitatory interneurons) are similar in their resting and action-potential mechanisms. Their most prominent differences are in size-related properties. The sensory-related interneurons, especially the edge cells and to some extent the giant interneurons, exhibited more pronounced differences in their resting and action-potential properties when compared with the other cell classes.