The role of endocannabinoid signaling in motor control.

Cannabinoid receptors and endocannabinoid signaling are distributed throughout the rostrocaudal neuraxis. Retrograde signaling via endocannabinoid mediates synaptic plasticity in many regions in the central nervous system. Here, we review the role of endocannabinoid signaling in different parts of the vertebrate motor system from networks responsible for the execution of movement to planning centers in the basal ganglia and cortex. The ubiquity of endocannabinoid-mediated plasticity suggests that it plays an important role in producing motion from defined circuitries and also for reconfiguring networks to learn new motor skills. The long-term plasticity induced by endocannabinoids may provide a long-term buffer that stabilizes the organization of motor circuits and their activity.

Transmitter phenotypes of commissural interneurons in the lamprey spinal cord.

The fundamental network for locomotion in all vertebrates contains a central pattern generator or CPG that produces the required motor output in the spinal cord. In the lamprey spinal cord different classes of interneuron's forming the core CPG circuitry have been characterized based on their morphological and electrophysiological features. The commissural interneuron's (C-INs) represent one essential component of CPG that have been implicated in controlling left-right alternation of the motor activity during swimming. However, it is still unclear if the C-INs displays a homogenous neurotransmitter phenotype and how they are distributed. In this paper we investigated the segmental distribution of glycine, glutamate and GABA-immunoreactive (ir) C-INs by combining retrograde Neurobiotin tracing with specific antibodies for these transmitters. The C-INs were more abundant in caudal and rostral segments adjacent to the injection site and their number gradually decreased in more distal segments, suggesting that these interneurons project over a short distance. The glycine-ir neurons represented around 50% of the total C-INs, while glutamate-ir neurons represented only 29%. Both types of C-INs were homogenously distributed over different segments along the spinal cord. Finally, no Neurobiotin labeled C-INs displayed GABA-ir, although many interneurons were ir to GABA, suggesting that GABAergic interneurons are not directly responsible for controlling left-right alternation of activity during locomotion in lamprey. Overall, these results show that the C-INs display a gradual rostrocaudal distribution and consist of both glycine- and glutamate-ir neurons. The difference in the proportion of inhibitory and excitatory C-INs represents an anatomical substrate that can ensure the predominance of alternating activity during locomotion.

Control of the temporal fidelity of synaptic transmission by a presynaptic high voltage-activated transient K current.

The type of K(+) channels controlling the waveform of the presynaptic spike and synaptic transmission were examined in the lamprey spinal cord. Reticulospinal neuron somata displayed a transient K(+) current with a high voltage-activation and inactivation. This current was selectively blocked by catechol at 100 microM. Reticulospinal axons also displayed a high voltage-activated fast K(+) current sensitive to catechol. The function of this presynaptic high voltage-activated fast K(+) current in controlling synaptic transmission was investigated by using paired intracellular recordings from reticulospinal axons and their targets. Blockade of this current by catechol (100 microM) prolonged the presynaptic spike elicited by a single stimulus leading to a potentiation of the postsynaptic EPSP. Calcium imaging of reticulospinal axons showed an increase in presynaptic calcium transients after blockade of the presynaptic K(+) current by catechol. During high frequency firing, catechol revealed an activity-dependent decrease in the spike duration, which resulted in a depression of synaptic transmission. These results suggest that the presynaptic high voltage-activated transient K(+) current acts to optimize the temporal fidelity of synaptic transmission by minimizing activity-dependent changes in the presynaptic spike waveform and calcium dynamics.

Marrow stromal cells form guiding strands in the injured spinal cord and promote recovery.

Marrow stromal cells (MSC) can be expanded rapidly in vitro and differentiated into multiple mesodermal cell types. In addition, differentiation into neuron-like cells expressing markers typical for mature neurons has been reported. To analyze whether such cells, exposed to differentiation media, could develop electrophysiological properties characteristic of neurons, we performed whole-cell recordings. Neuron-like MSC, however, lacked voltage-gated ion channels necessary for generation of action potentials. We then delivered MSC into the injured spinal cord to study the fate of transplanted MSC and possible effects on functional outcome in animals rendered paraplegic. MSC given 1 week after injury led to significantly larger numbers of surviving cells than immediate treatment and significant improvements of gait. Histology 5 weeks after spinal cord injury revealed that MSC were tightly associated with longitudinally arranged immature astrocytes and formed bundles bridging the epicenter of the injury. Robust bundles of neurofilament-positive fibers and some 5-hydroxytryptamine-positive fibers were found mainly at the interface between graft and scar tissue. MSC constitute an easily accessible, easily expandable source of cells that may prove useful in the establishment of spinal cord repair protocols.

Ion channels of importance for the locomotor pattern generation in the lamprey brainstem-spinal cord.

The intrinsic function of the spinal network that generates locomotion can be studied in the isolated brainstem-spinal cord of the lamprey, a lower vertebrate. The motor pattern underlying locomotion can be elicited in the isolated spinal cord. The network consists of excitatory glutamatergic and inhibitory glycinergic interneurones with known connectivity. The current review addresses the different subtypes of ion channels that are present in the cell types that constitute the network. In particular the roles of the different subtypes of Ca2+ channels and potassium channels that regulate integrated neuronal functions, like frequency regulation, spike frequency adaptation and properties that are important for generating features of the motor pattern (e.g. burst termination), are reviewed. By knowing the role of an ion channel at the cellular level, we also, based on previous knowledge of network connectivity, can understand which effect a given ion channel may exert at the different levels from molecule and cell to network and behaviour.

Characterization of a high-voltage-activated IA current with a role in spike timing and locomotor pattern generation.

Transient A-type K+ channels (I(A)) in neurons have been implicated in the delay of the spike onset and the decrease in the firing frequency. Here we have characterized biophysically and pharmacologically an I(A) current in lamprey locomotor network neurons that is activated by suprathreshold depolarization and is specifically blocked by catechol at 100 microM. The biophysical properties of this current are similar to the mammalian Kv3.4 channel. The role of the I(A) current both in single neuron firing and in locomotor pattern generation was analyzed. The I(A) current facilitates Na+ channel recovery from inactivation and thus sustains repetitive firing. The role of the I(A) current in motor pattern generation was examined by applying catechol during fictive locomotion induced by N-methyl-d-aspartate. Blockade of this current increased the locomotor burst frequency and decreased the firing of motoneurons. Although an alternating motor pattern could still be generated, the cycle duration was less regular, with ventral roots bursts failing on some cycles. Our results thus provide insights into the contribution of a high-voltage-activated I(A) current to the regulation of firing properties and motor coordination in the lamprey spinal cord.

Presynaptic inhibition and antidromic spikes in primary afferents of the crayfish: a computational and experimental analysis.

Primary afferent depolarizations (PADs) are associated with presynaptic inhibition and antidromic discharges in both vertebrates and invertebrates. In the present study, we have elaborated a realistic compartment model of a primary afferent from the coxobasipodite chordotonal organ of the crayfish based on anatomical and electrophysiological data. The model was used to test the validity of shunting and sodium channel inactivation hypotheses to account for presynaptic inhibition. Previous studies had demonstrated that GABA activates chloride channels located on the main branch close to the first branching point. We therefore focused the analysis on the effect of GABA synapses on the propagation of action potentials in the first axonal branch. Given the large diameters of the sensory axons in the region in which PADs were likely to be produced and recorded, the model indicates that a relatively large increase in chloride conductance (up to 300 nS) is needed to significantly reduce the amplitude of sensory spikes. The role of the spatial organization of GABA synapses in the sensory arborization was analyzed, demonstrating that the most effective location for GABA synapses is in the area of transition from active to passive conduction. This transition is likely to occur on the main branch a few hundred micrometers distal to the first branching point. As a result of this spatial organization, antidromic spikes generated by large-amplitude PADs are prevented from propagating distally.

Roles of high-voltage-activated calcium channel subtypes in a vertebrate spinal locomotor network.

Lamprey spinal cord neurons possess N-, L-, and P/Q-type high-voltage-activated (HVA) calcium channels. We have analyzed the role of the different HVA calcium channels subtypes in the overall functioning of the spinal locomotor network by monitoring the influence of their specific agonists and antagonists on synaptic transmission and on N-methyl-D-aspartate (NMDA)-elicited fictive locomotion. The N-type calcium channel blocker omega-conotoxin GVIA (omega-CgTx) depressed synaptic transmission from excitatory and inhibitory interneurons. Blocking L-type and P/Q-type calcium channels with nimodipine and omega-agatoxin, respectively, did not affect synaptic transmission. Application of omega-CgTx initially decreased the frequency of the locomotor rhythm, increased the burst duration, and subsequently increased the coefficient of variation and disrupted the motor pattern. These effects were accompanied by a depression of the synaptic drive between neurons in the locomotor network. Blockade of L-type channels by nimodipine also decreased the frequency and increased the duration of the locomotor bursts. Conversely, potentiation of L-type channels increased the frequency of the locomotor activity and decreased the duration of the ventral root bursts. In contrast to blockade of N-type channels, blockade or potentiation of L-type calcium channels had no effect on the stability of the locomotor pattern. The P/Q-type calcium channel blocker omega-agatoxin IVA had little effect on the locomotor frequency or burst duration. The results indicate that rhythm generation in the spinal locomotor network of the lamprey relies on calcium influx through L-type and N-type calcium channels.

Interaction between metabotropic and ionotropic glutamate receptors regulates neuronal network activity.

Experimental and computational techniques have been used to investigate the group I metabotropic glutamate receptor (mGluR)-mediated increase in the frequency of spinal cord network activity underlying locomotion in the lamprey. Group I mGluR activation potentiated the amplitude of NMDA-induced currents in identified motoneurons and crossed caudally projecting network interneurons. Group I mGluRs also potentiated NMDA-induced calcium responses. This effect was blocked by a group I mGluR-specific antagonist, but not by blockers of protein kinase A, C, or G. The effect of group I mGluRs activation was also tested on NMDA-induced oscillations known to occur during fictive locomotion. Activation of these receptors increased the duration of the plateau phase and decreased the duration of the hyperpolarizing phase. These effects were blocked by a group I mGluR antagonist. To determine its role in the modulation of NMDA-induced oscillations and the locomotor burst frequency, the potentiation of NMDA receptors by mGluRs was simulated using computational techniques. Simulating the interaction between these receptors reproduced the modulation of the plateau and hyperpolarized phases of NMDA-induced oscillations, and the increase in the frequency of the locomotor rhythm. Our results thus show a postsynaptic interaction between group I mGluRs and NMDA receptors in lamprey spinal cord neurons, which can account for the regulation of the locomotor network output by mGluRs.

Presynaptic inhibition and antidromic discharges in crayfish primary afferents.

The mechanisms of presynaptic inhibition have been studied in sensory afferents of a stretch receptor in an in vitro preparation of the crayfish. Axon terminals of these sensory afferents display primary afferent depolarisations (PADs) mediated by the activation of GABA receptors that open chloride channels. Intracellular labeling of sensory axons by Lucifer yellow combined with GABA immunohistochemistry revealed the presence of close appositions between GABA-immunoreactive boutons and sensory axons close to their first branching point within the ganglion. Electrophysiological studies showed that GABA inputs mediating PADs appear to occur around the first axonal branching point, which corresponds to the area of transition between active and passive propagation of spikes. Moreover, this study demonstrated that whilst shunting appeared to be the sole mechanism involved during small amplitude PADs, sodium channel inactivation occurred with larger amplitude PADs. However, when the largest PADs (>25 mV) are produced, the threshold for spike generation is reached and antidromic action potentials are elicited. The mechanisms involved in the initiation of antidromic discharges were analyzed by combining electrophysiological and simulation studies. Three mechanisms act together to ensure that PAD-mediated spikes are not conveyed distally: 1) the lack of active propagation in distal regions of the sensory axons; 2) the inactivation of the sodium channels around the site where PADs are produced; and 3) a massive shunting through the opening of chloride channels associated with the activation of GABA receptors. The centrally generated spikes are, however, conveyed antidromically in the sensory nerve up to the proprioceptive organ, where they inhibit the activity of the sensory neurons for several hundreds of milliseconds.

GABA(B) receptor activation inhibits N- and P/Q-type calcium channels in cultured lamprey sensory neurons.

In lamprey, sensory transmission from mechanosensory receptors (dorsal cells) to central neurons is presynaptically inhibited by GABA(B) receptor activation. The mechanisms underlying this effect were investigated using isolated dorsal cells, where voltage-dependent calcium currents were recorded in the whole-cell configuration. Activation of GABA(B) receptors by baclofen decreased the peak amplitude of high voltage-activated (HVA) calcium currents and slowed the activation phase. The role of G-proteins in mediating the effects of baclofen was examined. Intracellular dialysis of GTPgammaS occluded the effects of baclofen. Intracellular dialysis of GDPbetaS and preincubation in pertussis toxin both attenuated the effect of baclofen. Specific calcium channel blockers were used to study the types of HVA calcium channels involved in the GABA(B)-mediated modulation. The baclofen-induced inhibition was not affected by the L-type calcium channel antagonist nimodipine, but was partially blocked by the N-type blocker omega-conotoxin GVIA, and completely occluded by omega-conotoxin MVIIC, a blocker of both N- and P/Q-type channels. The pharmacology of dorsal cell GABA(B) receptors was studied using two agonists, baclofen and CGP 27492, and four antagonists, CGP 35348, CGP 55845, phaclofen and saclofen. The inhibition induced by either of the two agonists was blocked by CGP 55845, phaclofen and saclofen. The antagonist CGP 35348 completely blocked the inhibition of HVA calcium current induced by the agonist CGP 27492, but had no effect on baclofen-induced GABA(B) receptor activation. This study thus demonstrates that GABA(B) receptor activation in lamprey mechanosensory neurons inhibits N- and P/Q-type calcium channels in a voltage- and G-protein-dependent manner.

Inhibition of N- and L-type Ca2+ currents by dopamine in lamprey spinal motoneurons.

Dopamine is co-localized with 5-hydroxytryptamine in a ventromedially located plexus in the lamprey spinal cord and reduces Ca2+ currents in motoneurons that express high-voltage activated Ca2+ currents of the N-, L- and P/Q-types. Blockade of L- and P/Q- type channels leaving N-type channels intact reduced but did not prevent the inhibition of the Ca2+ current by dopamine. Dopamine also reduced the L-type current potentiated by BAY K 8644. During simultaneous blockade of N-type and L-type currents, dopamine was unable to affect the remaining Ca2+ current. In addition, blockade of G-proteins abolished the dopaminergic modulation. The inhibition was unaffected by depolarizing prepulses. Thus, dopamine mediates inhibition of N- and L-type currents through a G-protein-dependent, voltage-independent pathway in lamprey spinal motoneurons.

Shunting versus inactivation: analysis of presynaptic inhibitory mechanisms in primary afferents of the crayfish.

Primary afferent depolarizations (PADs) are associated with presynaptic inhibition in both vertebrates and invertebrates. In the present study, we have used both anatomical and electrophysiological techniques to analyze the relative importance of shunting mechanisms versus sodium channel inactivation in mediating the decrease of action potential amplitude, and thereby presynaptic inhibition. Experiments were performed in sensory afferents of a stretch receptor in an in vitro preparation of the crayfish. Lucifer yellow intracellular labeling of sensory axons combined with GABA immunohistochemistry revealed close appositions between GABA-immunoreactive (ir) fibers and sensory axons. Most contacts were located on the main axon at the entry zone of the ganglion, close to the first branching point within the ganglion. By comparison, the output synapses of sensory afferents to target neurons were located on distal branches. The location of synaptic inputs mediating spontaneous PADs was also determined electrophysiologically by making dual intracellular recordings from single sensory axons. Inputs generating PADs appear to occur around the first axonal branching point, in agreement with the anatomical data. In this region, small PADs (3-15 mV) produced a marked reduction of action potential amplitude, whereas depolarization of the membrane potential by current injection up to 15 mV had no effect. These results suggest that the decrease of the amplitude of action potentials by single PADs results from a shunting mechanism but does not seem to involve inactivation of sodium channels. Our results also suggest that GABAergic presynaptic inhibition may act as a global control mechanism to block transmission through certain reflex pathways.

Calcium channels involved in synaptic transmission from reticulospinal axons in lamprey.

The pharmacology of calcium channels involved in glutamatergic synaptic transmission from reticulospinal axons in the lamprey spinal cord was analyzed with specific agonists and antagonists of different high-voltage activated calcium channels. The N-type calcium channel blocker omega-conotoxin GVIA (omega-CgTx) induced a large decrease of the amplitude of reticulospinal-evoked excitatory postsynaptic potentials (EPSPs). The P/Q-type calcium channel blocker omega-agatoxin IVA (omega-Aga) also reduced the amplitude of the reticulospinal EPSPs, but to a lesser extent than omega-CgTx. The dihydropyridine agonist Bay K and antagonist nimodipine had no effect on the amplitude of the reticulospinal EPSP. Combined application of omega-CgTx and omega-Aga strongly decreased the amplitude the EPSPs but was never able to completely block them, indicating that calcium channels insensitive to these toxins (R-type) are also involved in synaptic transmission from reticulospinal axons. We have previously shown that the group III metabotropic glutamate receptor agonist L(+)-2-amino-4-phosphonobutyric acid (L-AP4) mediates presynaptic inhibition at the reticulospinal synapse. To test if this presynaptic effect is mediated through inhibition of calcium influx, the effect of L-AP4 on reticulospinal transmission was tested before and after blockade of N-type channels, which contribute predominantly to transmitter release at this synapse. Blocking the N-type channels with omega-CgTx did not prevent inhibition of reticulospinal synaptic transmission by L-AP4. In addition, L-AP4 had no affect on the calcium current recorded in the somata of reticulospinal neurons or on the calcium component of action potentials in reticulospinal axons. These results show that synaptic transmission from reticulospinal axons in the lamprey is mediated by calcium influx through N-, P/Q- and R-type channels, with N-type channels playing the major role. Furthermore, presynaptic inhibition of reticulospinal transmission by L-AP4 appears not to be mediated through inhibition of presynaptic calcium channels.

Endogenous activation of metabotropic glutamate receptors contributes to burst frequency regulation in the lamprey locomotor network.

The effect of metabotropic glutamate receptor (mGluR) agonists and antagonists on the spinal cord network underlying locomotion in the lamprey has been analysed. The specific group I mGluR agonist (R,S)-3,5-dihydroxyphenylglycine (DHPG) and the broad-spectrum mGluR agonist (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD) both increased the burst frequency of N-methyl-D-aspartic acid (NMDA)-induced fictive locomotion and depolarized grey matter neurons. The burst frequency increase induced by the mGluR agonists was counteracted by the mGluR antagonists (+)-alpha-methyl-4-carboxyphenylglycine ((+)-MCPG), cyclopropan[b]chromen-1a-carboxylic acid ethylester (CPCCOEt) and (RS)-1-aminoindan-1,5-dicarboxylic acid (AIDA). Application of CPCCOEt alone reduced the locomotor burst frequency, indicating that mGluRs are endogenously activated during fictive locomotion. The mGluR antagonist CPCCOEt had no effect on NMDA-, or (S)-alpha-amino-3-hydroxy-5-methyl-4-isoazolepropionic acid (AMPA)-induced depolarizations. The mGluR agonists 1S,3R-ACPD and DHPG increased the amplitude of NMDA-induced depolarizations, a mechanism which could account for the increase in burst frequency. The group III mGluR agonist L-2-amino-4-phosphonobutyric acid reduced intraspinal synaptic transmission and burst frequency.

Calcium influx through N- and P/Q-type channels activate apamin-sensitive calcium-dependent potassium channels generating the late afterhyperpolarization in lamprey spinal neurons.

Lamprey spinal neurons exhibit a fast afterhyperpolarization and a late afterhyperpolarization (AHP) which is due to the activation of apamin-sensitive SK Ca2+-dependent K+ channels (KCa) activated by calcium influx through voltage-dependent channels during the action potential (Hill et al. 1992, Neuroreport, 3, 943-945). In this study we have investigated which calcium channel subtypes are responsible for the activation of the KCa channels underlying the AHP. The effects of applying specific calcium channel blockers and agonists were analysed with regard to their effects on the AHP. Blockade of N-type calcium channels by omega-conotoxin GVIA resulted in a significant decrease in the amplitude of the AHP by 76.2+/-14.9% (mean +/- SD). Application of the P/Q-type calcium channel blocker omega-agatoxin IVA reduced the amplitude of the AHP by 20.3+/-10.4%. The amplitude of the AHP was unchanged during application of the L-type calcium channel antagonist nimodipine or the agonist (+/-)-BAY K 8644, as was the compound afterhyperpolarization after a train of 10 spikes at 100 Hz. The effects of calcium channel blockers were also tested on the spike frequency adaptation during a train of action potentials induced by a 100-200 ms depolarizing pulse. The N- and P/Q-type calcium channel antagonists decreased the spike frequency adaptation, whereas blockade of L-type channels had no effect. Thus in lamprey spinal cord motor- and interneurons, apamin-sensitive KCa channels underlying the AHP are activated primarily by calcium entering through N-type channels, and to a lesser extent through P/Q-type channels.

Intrinsic function of a neuronal network - a vertebrate central pattern generator.

The cellular bases of vertebrate locomotor behaviour is reviewed using the lamprey as a model system. Forebrain and brainstem cell populations initiate locomotor activity via reticulospinal fibers activating a spinal network comprised of glutamatergic and glycinergic interneurons. The role of different subtypes of Ca2+ channels, Ca2+ dependent K+ channels and voltage dependent NMDA channels at the neuronal and network level is in focus as well as the effects of different metabotropic, aminergic and peptidergic modulators that target these ion channels. This is one of the few vertebrate networks that is understood at a cellular level.

Vertebrate locomotion--a lamprey perspective.

The forebrain, brain stem, and spinal cord contribution to the control of locomotion is reviewed in this chapter. The lamprey is used as an experimental model because it allows a detailed cellular analysis of the neuronal network underlying locomotion. The focus is on cellular mechanisms that are important for the pattern generation, as well as different types of pre- and postsynaptic modulation. Neuropeptides target different cellular and synaptic mechanisms and cause long-lasting changes (> 24 h) in network function.

Calcium channel subtypes in lamprey sensory and motor neurons.

Pharmacologically distinct calcium channels have been characterized in dissociated cutaneous sensory neurons and motoneurons of the larval lamprey spinal cord. To enable cell identification, sensory dorsal cells and motoneurons were selectively labeled with fluorescein-coupled dextran amine in the intact spinal cord in vitro before dissociation. Calcium channels present in sensory dorsal cells, motoneurons, and other spinal cord neurons were characterized with the use of whole cell voltage-clamp recordings and specific calcium channel agonist and antagonists. The results show that a transient low-voltage-activated (LVA) calcium current was present in a proportion of sensory dorsal cells but not in motoneurons, whereas high-voltage-activated (HVA) calcium currents were seen in all neurons recorded. The different components of HVA current were dissected pharmacologically and similar results were obtained for both dorsal cells and motoneurons. The N-type calcium channel antagonist omega-conotoxin-GVIA (omega-CgTx) blocked >70% of the HVA current. A large part of the omega-CgTx block was reversed after washout of the toxin. The L-type calcium channel antagonist nimodipine blocked approximately 15% of the total HVA current. The dihydropyridine agonist (+/-)-BayK 8644 markedly increased the amplitude of the calcium channel current. The BayK-potentiated current was not affected by omega-CgTx, indicating that the reversibility of the omega-CgTx effect is not due to a blockade of L-type channels. Simultaneous application of omega-CgTx and nimodipine left approximately 15% of the HVA calcium channel current, a small part of which was blocked by the P/Q-type channel antagonist omega-agatoxin-IVA. In the presence of the three antagonists, the persistent residual current (approximately 10%) was completely blocked by cadmium. Our results provide evidence for the existence of HVA calcium channels of the N, L, and P/Q types and other HVA calcium channels in lamprey sensory neurons and motoneurons. In addition, certain types of neurons express LVA calcium channels.

Afferents of the lamprey striatum with special reference to the dopaminergic system: a combined tracing and immunohistochemical study.

The origin of afferents to the striatum in lamprey (Lampetra fluviatilis) was studied by using fluorescein-coupled dextran-amines (FDA). Injection of FDA into the striatum retrogradely labeled several cell populations in the forebrain and the rostral rhombencephalon. No retrograde labeled cells were seen in the mesencephalon. A dopamine-specific antiserum was used to determined the distribution of dopaminergic perikarya and fibers. Many dopamine-immunoreactive (DA-ir) fibers were present throughout the brain, but the highest density of labeled fibers was in the mediobasal prosencephalon, especially in the striatum, the lateral hypothalamic area, and the neurohypophysis. Most DA-ir cells were located in the mediobasal diencephalon (preoptic region, nucleus commissurae postopticae, hypothalamus, and nucleus tuberculi posterioris). In the mesencephalon, only a few immunopositive cells were observed in the tectum opticum. In the rhombencephalon, DA-ir cells were observed in the isthmic region, dorsally to the descending trigeminal tract, and caudally to the posterior rhombencephalic reticular nucleus. The rostralmost spinal cord received many descending DA-ir fibers from the brainstem. Along the spinal cord, DA-ir neurons were also found, some of which projected to the medioventral surface, forming a prominent plexus. On the basis of double-labeling experiments, it is shown that the dopaminergic input to the striatum originates from the nucleus tuberculi posterioris. Thus, the striatum receives inputs from different structures, including a strong dopaminergic innervation from the diencephalon. Much of the dopaminergic system in Lampetra fluviatilis is basically similar to that seen in some teleosts, but it presents differences with other anamniote (elasmobranch) as well as amniote groups.