James T. Buchanan

Contributions of Identifiable Neurons and Neuron Classes to Lamprey Vertebrate Neurobiology

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.

Neural network simulations of coupled locomotor oscillators in the lamprey spinal cord

The segmental locomotor network in the lamprey spinal cord was simulated on a computer using a connectionist-type neural network. The cells of the network were identical except for their excitatory levels and their synaptic connections. The synaptic connections used were based on previous experimental work. It was demonstrated that the connectivity of the circuit is capable of generating oscillatory activity with the appropriate phase relations among the cells. Intersegmental coordination was explored by coupling two identical segmental networks using only the cells of the network. Each of the possible couplings of a bilateral pair of cells in one oscillator with a bilateral pair of cells in the other oscillator produced stable phase locking of the two oscillators. The degree of phase difference was dependent upon synaptic weight, and the operating range of synaptic weights varied among the pairs of connections. The coupling was tested using several criteria from experimental work on the lamprey spinal cord. Coupling schemes involving several pairs of connecting cells were found which 1) achieved steadystate phase locking within a single cycle, 2) exhibited constant phase differences over a wide range of cycle periods, and 3) maintained stable phase locking in spite of large differences in the intrinsic frequencies of the two oscillators. It is concluded that the synaptic connectivity plays a large role in producing oscillations in this network and that it is not necessary to postulate a separate set of coordinating neurons between oscillators in order to achieve appropriate phase coupling.

5-Hydroxytryptamine depresses reticulospinal excitatory postsynaptic potentials in motoneurons of the lamprey

Application of 5-hydroxytryptamine (5-HT) to the lamprey spinal cord in vitro reversibly depressed the chemical component of excitatory post-synaptic potentials recorded intracellularly in motoneurons and evoked by stimulation of single reticulospinal Müller cells. The depression could be produced either by local application of small volumes of 10 mM 5-HT to the surface of the spinal cord or by bath-application of 1 or 10 microM 5-HT. No effect on the input resistance of the postsynaptic cells or their sensitivity to glutamate, the suspected transmitter at this synapse, could be detected, suggesting the possibility of a presynaptic action of 5-HT at this synapse in the lamprey.

Neuronal control of swimming behavior: comparison of vertebrate and invertebrate model systems.

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 40 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, neural-circuitry, 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.

Apamin reduces the late afterhyperpolarization of lamprey spinal neurons, with little effect on fictive swimming

The role of the late afterhyperpolarization (late AHP) in the firing properties of lamprey spinal neurons was tested by bath application of apamin, a selective blocker of the sk calcium-dependent potassium current. Intracellular recordings of identified motoneurons and interneurons were made with micropipette electrodes in the isolated lamprey spinal cord. Apamin reversibly reduced the amplitude of the late afterhyperpolarization without affecting other aspects of the action potential or the resting potential. The firing frequencies of the neurons were enhanced by apamin over a range of depolarizing current pulse injections. The effect of apamin was also tested on fictive swimming, which was induced in the isolated spinal cord by bath application of an excitatory amino acid (D-glutamate or N-methyl-D,L-aspartate). A concentration of apamin (10 microM) sufficient to substantially reduce the late AHP had no significant effect on the ventral root burst rate, intensity, or phase lag during fictive swimming.

THE NEURONAL NETWORK FOR LOCOMOTION IN THE LAMPREY SPINAL CORD : EVIDENCE FOR THE INVOLVEMENT OF COMMISSURAL INTERNEURONS

The spinal cord of the lamprey, a primitive vertebrate, has been used as a model system for investigating the cellular basis of rhythmic locomotor activity. Three classes of interneurons have been characterized that are active during locomotor activity in the isolated spinal cord (ie fictive swimming). The identified synaptic interactions of these neurons form a network which has been proposed to underlie locomotor rhythmogenesis. Modeling studies confirmed that the network can produce oscillatory activity with phase relations among the neurons similar to those found in the spinal cord. Within the network, inhibitory commissural interneurons form reciprocal inhibitory connections and play a key role in rhythmogenesis. Several experiments have been done to test whether these cells participate in the generation of rhythmic activity in the spinal cord. First, midline lesions that sever the axons of commissural interneurons eliminate rhythmic ventral root bursting. Second, photo-ablation of commissural interneurons on one side of the spinal cord alters the symmetry of ventral root bursts, alters the cycle period, and can eliminate rhythmic bursting. Taken together, these experiments support the model that commissural interneurons are involved in rhythmogenesis in the lamprey spinal cord.

Effects of strychnine on fictive swimming in the lamprey: evidence for glycinergic inhibition, discrepancies with model predictions, and novel modulatory rhythms

Abstract1.Inhibitory postsynaptic potentials (ipsps) produced by two classes of interneurons, CC (contralateral and caudal projecting) and lateral interneurons, were tested for strychnine sensitivity using paired intracellular recordings in the lamprey spinal cord. The ipsps were partially blocked by 0.2–0.5 μM strychnine and were completely blocked by 5 μM strychnine. Thus, the ipsps may be glycinergic.2.These interneurons are key participants in a proposed circuit model for fictive swimming. A connectionisttype computer simulation of the model demonstrated that the cycle period of the network increased with decreasing ipsp strength.3.Application of strychnine (0.1–0.5 μM) to the spinal cord during fictive swimming induced by an excitatory amino acid increased cycle period, consistent with previous reports, but at odds with stimulation predictions.4.Strychnine also produced slow rhythmic modulation of fictive swimming (period = 12 s) which maintained left-right alternation and rostral-caudal coordination. Auto- and cross-correlation analyses revealed that the slow modulation was present in a weaker form in most control preparations during fictive swimming.5.Since the proposed model for the swimming pattern generator in the lamprey spinal cord does not predict the observed speeding with strychnine, nor the slow modulatory rhythm, it appears to be deficient in its present formulation.

Chapter 27 The Roles of Spinal Interneurons and Motoneurons in the Lamprey Locomotor Network

The isolated lamprey spinal cord offers a relatively simple and convenient adult preparation in which to investigate how nerve cells generate behavior and in particular the rhythmic motor patterns of locomotion. Nerve cell classes can be identified and their cellular and synaptic properties characterized, and a simple model based on demonstrated synaptic connectivity can account for major aspects of fictive swimming. Clearly, however, much remains to be learned. In particular, the properties of the spinal neurons have been shown to change during swimming activity but relatively little is known about how these changes occur or the effects that these changes have upon the activities of the network. In addition, much remains to be learned about the cell types and their synaptic interactions as demonstrated here with the newly discovered feedback connections from motoneurons, which have not been previously taken into account in modeling of the lamprey locomotor network.

Modulation of swimming in the lamprey, Petromyzon marinus, by serotonergic and dopaminergic drugs

The effects of serotonergic and dopaminergic drugs on free swimming behavior in adult sea lampreys (Petromyzon marinus) were investigated using video image analysis. Injections of the serotonin precursor 5-hydroxy-L-tryptophan along with the serotonin reuptake blocker clomipramine into the visceral cavity of lampreys resulted in significant increases in the cycle period of swimming, but had no significant effects on the propagation time of the swim waves down the body (normalized to cycle period), or on the degree of body curvature. Injections of the dopamine agonist apomorphine resulted in significant decreases of cycle period and body curvature with no significant effects on the normalized wave propagation time. The effects on cycle period are consistent with previous findings using serotonin and apomorphine on swimming activity in the isolated spinal cord.

Quantitative analysis of electrotonic structure and membrane properties of nmda-activated lamprey spinal neurons

Parameter optimization methods were used to quantitatively analyze frequency-domain-voltage-clamp data of NMDA-activated lamprey spinal neurons simultaneously over a wide range of membrane potentials. A neuronal cable model was used to explicitly take into account receptors located on the dendritic trees. The driving point membrane admittance was measured from the cell soma in response to a Fourier synthesized point voltage clamp stimulus. The data were fitted to an equivalent cable model consisting of a single lumped soma compartment coupled resistively to a series of equal dendritic compartments. The model contains voltage-dependent NMDA sensitive (INMDA), slow potassium (IK), and leakage (IL) currents. Both the passive cable properties and the voltage dependence of ion channel kinetics were estimated, including the electrotonic structure of the cell, the steady-state gating characteristics, and the time constants for particular voltage- and time-dependent ionic conductances. An alternate kinetic formulation was developed that consisted of steady-state values for the gating parameters and their time constants at half-activation values as well as slopes of these parameters at half-activation. This procedure allowed independent restrictions on the magnitude and slope of both the steady-state gating variable and its associated time constant. Quantitative estimates of the voltage-dependent membrane ion conductances and their kinetic parameters were used to solve the nonlinear equations describing dynamic responses. The model accurately predicts current clamp responses and is consistent with experimentally measured TTX-resistant NMDA-induced patterned activity. In summary, an analysis method is developed that provides a pragmatic approach to quantitatively describe a nonlinear neuronal system.