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Chemoreception plays a central role in the survival and reproduction of many animals in aquatic environments. Fishes use three chemosensory systems, the olfactory system, the diffuse chemosensory system, and the taste (gustatory) system. While these are present in the sea lamprey, a majority of chemosensory research has focused on olfaction. This review summarizes current knowledge of the lamprey chemosensory systems, including stimuli, receptors, neural pathways and behavioural roles of the chemosensory stimuli. A description of the olfactory sensory neurons located deep within the nostril, of synaptic pathways within the brain and of modulatory mechanisms that underly the translation of olfaction to movement are included. The lamprey diffuse chemosensory system may enable solitary chemosensory cells on the skin of the mouth, nostril, gill, and tailfin to function as sentinels for rapid responses to chemical stimuli. While little is known abouth the lamprey taste system, taste buds are located within the pharynx, thus enabling chemosensory responses to material that has entered the body cavity. Knowledge and understanding of function of these three chemosensory systems supports the management of sea lamprey populations.

The olfactory system allows animals to navigate in their environment to feed, mate, and escape predators. It is well established that odorant exposure or electrical stimulation of the olfactory system induces stereotyped motor responses in fishes. However, the neural circuitry responsible for the olfactomotor transformations is only beginning to be unraveled. A neural substrate eliciting motor responses to olfactory inputs was identified in the lamprey, a basal vertebrate used extensively to examine the neural mechanisms underlying sensorimotor transformations. Two pathways were discovered from the olfactory organ in the periphery to the brainstem motor nuclei responsible for controlling swimming. The first pathway originates from sensory neurons located in the accessory olfactory organ and reaches a single population of projection neurons in the medial olfactory bulb, which, in turn, transmit the olfactory signals to the posterior tuberculum and then to downstream brainstem locomotor centers. A second pathway originates from the main olfactory epithelium and reaches the main olfactory bulb, the neurons of which project to the pallium/cortex. The olfactory signals are then conveyed to the posterior tuberculum and then to brainstem locomotor centers. Olfactomotor behavior can adapt, and studies were aimed at defining the underlying neural mechanisms. Modulation of bulbar neural activity by GABAergic, dopaminergic, and serotoninergic inputs is likely to provide strong control over the hardwired circuits to produce appropriate motor behavior in response to olfactory cues. This review summarizes current knowledge relative to the neural circuitry producing olfactomotor behavior in lampreys and their modulatory mechanisms.

Locomotion is a basic motor act essential for survival. Amongst other things, it allows animals to move in their environment to seek food, escape predators, or seek mates for reproduction. The neural mechanisms involved in the control of locomotion have been examined in many vertebrate species and a clearer picture is progressively emerging. The basic muscle synergies responsible for propulsion are generated by neural networks located in the spinal cord. In turn, descending supraspinal inputs are responsible for starting, maintaining, and stopping locomotion as well as for steering and controlling speed. Several neurotransmitter systems play a crucial role in modulating the neural activity during locomotion. For instance, cholinergic inputs act both at the spinal and supraspinal levels and the underlying mechanisms are the focus of the present review. Much information gained on supraspinal cholinergic modulation of locomotion was obtained from the lamprey model. Nicotinic cholinergic inputs increase the level of excitation of brainstem descending command neurons, the reticulospinal neurons (RSNs), whereas muscarinic inputs activate a select group of hindbrain neurons that project to the RSNs to boost their level of excitation. Muscarinic inputs also reduce the transmission of sensory inputs in the brainstem, a phenomenon that could help in sustaining goal directed locomotion. In the spinal cord, intrinsic cholinergic inputs strongly modulate the activity of interneurons and motoneurons to control the locomotor output. Altogether, the present review underlines the importance of the cholinergic inputs in the modulation of locomotor activity in vertebrates.

Locomotion in vertebrates relies on high brain centers converging onto the mesencephalic locomotor region (MLR). How the MLR recruits brainstem reticulospinal neurons (RSNs) to initiate locomotion is incompletely understood due to the challenge of recording these cells in vivo. To tackle this question, we leveraged the transparency and genetic accessibility of larval zebrafish. In this model organism, we uncovered the locus of the MLR as a small region dorsal to the locus coeruleus containing glutamatergic and cholinergic neurons. MLR stimulations reliably elicited forward bouts of controlled duration and speed. We find that the MLR elicits forward locomotion by recruiting V2a RSNs in the pontine and retropontine regions, and gradually in the medulla. Remarkably, recruited V2a RSNs in the medulla act as maintain cells encoding speed of forward locomotion. Altogether, our study reveals that the MLR recruits genetically-identified reticulospinal neurons in the medulla to control the kinematics of exploration.

Molecules present in an animal's environment can indicate the presence of predators, food, or sexual partners and consequently, induce migratory, reproductive, foraging, or escape behaviors. Three sensory systems, the olfactory, gustatory, and solitary chemosensory cell (SCC) systems detect chemical stimuli in vertebrates. While a great deal of research has focused on the olfactory and gustatory system over the years, it is only recently that significant attention has been devoted to the SCC system. The SCCs are microvillous cells that were first discovered on the skin of fish, and later in amphibians, reptiles, and mammals. Lampreys also possess SCCs that are particularly numerous on cutaneous papillae. However, little is known regarding their precise distribution, innervation, and function. Here, we show that sea lampreys (Petromyzon marinus L.) have cutaneous papillae located around the oral disk, nostril, gill pores, and on the dorsal fins and that SCCs are particularly numerous on these papillae. Tract-tracing experiments demonstrated that the oral and nasal papillae are innervated by the trigeminal nerve, the gill pore papillae are innervated by branchial nerves, and the dorsal fin papillae are innervated by spinal nerves. We also characterized the response profile of gill pore papillae to some chemicals and showed that trout-derived chemicals, amino acids, and a bile acid produced potent responses. Together with a companion study (Suntres et al., Journal of Comparative Neurology, this issue), our results provide new insights on the function and evolution of the SCC system in vertebrates.

Animals have developed different locomotor strategies to explore and survive in their environment by walking, flying, or swimming. Even though the biomechanics are different, the neural control of these movements is very similar in all vertebrate species. In this review, we provide an overview of the descending central control of locomotion in vertebrates with an emphasis on recent findings in the field. We discuss how different cell populations in the spinal cord control the intensity of locomotion. We then outline exciting findings on the heterogeneity of reticulospinal cells and their role in controlling different locomotor aspects. Furthermore, we review specific roles of different cell groups in the mesencephalic locomotor region (MLR) and describe newly found descending inputs to the MLR.

Locomotion occurs sporadically and needs to be started, maintained, and stopped. The neural substrate underlying the activation of locomotion is partly known, but little is known about mechanisms involved in termination of locomotion. Recently, reticulospinal neurons (stop cells) were found to play a crucial role in stopping locomotion in the lamprey: their activation halts ongoing locomotion and their inactivation slows down the termination process. Intracellular recordings of these cells revealed a distinct activity pattern, with a burst of action potentials at the beginning of a locomotor bout and one at the end (termination burst). The termination burst was shown to be time linked to the end of locomotion, but the mechanisms by which it is triggered have remained unknown. We studied this in larval sea lampreys (Petromyzon marinus; the sex of the animals was not taken into account). We found that the mesencephalic locomotor region (MLR), which is known to initiate and control locomotion, stops ongoing locomotion by providing synaptic inputs that trigger the termination burst in stop cells. When locomotion is elicited by MLR stimulation, a second MLR stimulation stops the locomotor bout if it is of lower intensity than the initial stimulation. This occurs for MLR-induced, sensory-evoked, and spontaneous locomotion. Furthermore, we show that glutamatergic and, most likely, monosynaptic projections from the MLR activate stop cells during locomotion. Therefore, activation of the MLR not only initiates locomotion, but can also control the end of a locomotor bout. These results provide new insights onto the neural mechanisms responsible for stopping locomotion.SIGNIFICANCE STATEMENTThe mesencephalic locomotor region (MLR) is a brainstem region well known to initiate and control locomotion. Since its discovery in cats in the 1960s, the MLR has been identified in all vertebrate species tested from lampreys to humans. We now demonstrate that stimulation of the MLR not only activates locomotion, but can also stop it. This is achieved through a descending glutamatergic signal, most likely monosynaptic, from the MLR to the reticular formation that activates reticulospinal stop cells. Together, our findings have uncovered a neural mechanism for stopping locomotion and bring new insights into the function of the MLR.

Lampreys are basal vertebrates with a nervous system very similar to that of mammals but with a much smaller size and far fewer neurons. The lamprey nervous system can be entirely maintained in vitro for extended periods of time (easily up to 40 h). For these reasons, the lamprey has been considered as a model of choice to characterize the neural mechanisms underlying locomotion at the system, cellular, and subcellular levels since its introduction in the late 1970s by Sten Grillner and his collaborators in Sweden. This chapter summarizes some of the salient discoveries made in lampreys and examines how this animal model will likely be the source of future exciting discoveries on the neural control of locomotion.

Meso-diencephalic dopaminergic neurons are known to modulate locomotor behaviors through their ascending projections to the basal ganglia, which in turn project to the mesencephalic locomotor region, known to control locomotion in vertebrates. In addition to their ascending projections, dopaminergic neurons were found to increase locomotor movements through direct descending projections to the mesencephalic locomotor region and spinal cord. Intriguingly, fibers expressing tyrosine hydroxylase (TH), the rate-limiting enzyme of dopamine synthesis, were also observed around reticulospinal neurons of lampreys. We now examined the origin and the role of this innervation. Using immunofluorescence and tracing experiments, we found that fibers positive for dopamine innervate reticulospinal neurons in the four reticular nuclei of lampreys. We identified the dopaminergic source using tracer injections in reticular nuclei, which retrogradely labeled dopaminergic neurons in a caudal diencephalic nucleus (posterior tuberculum [PT]). Using voltammetry in brain preparations isolatedin vitro, we found that PT stimulation evoked dopamine release in all four reticular nuclei, but not in the spinal cord. In semi-intact preparations where the brain is accessible and the body moves, PT stimulation evoked swimming, and injection of a D1receptor antagonist within the middle rhombencephalic reticular nucleus was sufficient to decrease reticulospinal activity and PT-evoked swimming. Our study reveals that dopaminergic neurons have access to command neurons that integrate sensory and descending inputs to activate spinal locomotor neurons. As such, our findings strengthen the idea that dopamine can modulate locomotor behavior both via ascending projections to the basal ganglia and through descending projections to brainstem motor circuits.SIGNIFICANCE STATEMENTMeso-diencephalic dopaminergic neurons play a key role in modulating locomotion by releasing dopamine in the basal ganglia, spinal networks, and the mesencephalic locomotor region, a brainstem region that controls locomotion in a graded fashion. Here, we report in lampreys that dopaminergic neurons release dopamine in the four reticular nuclei where reticulospinal neurons are located. Reticulospinal neurons integrate sensory and descending suprareticular inputs to control spinal interneurons and motoneurons. By directly modulating the activity of reticulospinal neurons, meso-diencephalic dopaminergic neurons control the very last instructions sent by the brain to spinal locomotor circuits. Our study reports on a new direct descending dopaminergic projection to reticulospinal neurons that modulates locomotor behavior.

Detection of chemical cues is important to guide locomotion in association with feeding and sexual behavior. Two neural pathways responsible for odor‐evoked locomotion have been characterized in the sea lamprey (Petromyzon marinus L.), a basal vertebrate. There is a medial pathway originating in the medial olfactory bulb (OB) and a lateral pathway originating from the rest of the OB. These olfactomotor pathways are present throughout the life cycle of lampreys, but olfactory‐driven behaviors differ according to the developmental stage. Among possible mechanisms, dopaminergic (DA) modulation in the OB might explain the behavioral changes. Here, we examined DA modulation of olfactory transmission in lampreys. Immunofluorescence against DA revealed immunoreactivity in the OB that was denser in the medial part (medOB), where processes were observed close to primary olfactory afferents and projection neurons. Dopaminergic neurons labeled by tracer injections in the medOB were located in the OB, the posterior tuberculum, and the dorsal hypothalamic nucleus, suggesting the presence of both intrinsic and extrinsic DA innervation. Electrical stimulation of the olfactory nerve in an in vitro whole‐brain preparation elicited synaptic responses in reticulospinal cells that were modulated by DA. Local injection of DA agonists in the medOB decreased the reticulospinal cell responses whereas the D2 receptor antagonist raclopride increased the response amplitude. These observations suggest that DA in the medOB could modulate odor‐evoked locomotion. Altogether, these results show the presence of a DA innervation within the medOB that may play a role in modulating olfactory inputs to the motor command system of lampreys., Dopaminergic innervation in the medial olfactory bulb was shown to modulate olfactory inputs to the motor command system. Dopaminergic processes were anatomically traced back to local and diencephalic DA neurons. Furthermore, microinjection of DA agonists in the medOB decreased reticulospinal cell responses to olfactory stimulation.

Solitary chemosensory cells (SCCs) and their innervating fibers are located in the respiratory system of many vertebrates, including papillae on lamprey gill pores. In order to gain stronger insight for the role of these chemosensory cells, we examined immunocytochemical and innervation characteristics, as well as abundance at the different stages of the lamprey life cycle. The SCCs were distinguished from the surrounding epithelial cells by calretinin and phospholipase C140 immunoreactivity. Nerve fibers extended into the gill pore papillae, as far as the SCCs and serotonergic fibers extended from the underlying dermis into the papillar base. Gill pore papillae were absent and SCCs were sparse during the larval stage and in newly transformed lamprey. Few SCCs were located on small nub-like papillae during the parasitic juvenile stage, but SCCs were abundant on prominent papillae in migrating and in spawning adults. These findings show similarities between the SCCs in lampreys and other vertebrates and suggest that gill SCC function may be important during the feeding juvenile and the adult stages of the lamprey life cycle.

SummaryLocomotion requires the proper sequencing of neural activity to start, maintain, and stop it. Recently, brainstem neurons were shown to specifically stop locomotion in mammals. However, the cellular properties of these neurons and their activity during locomotion are still unknown. Here, we took advantage of the lamprey model to characterize the activity of a cell population that we now show to be involved in stopping locomotion. We find that these neurons display a burst of spikes that coincides with the end of swimming activity. Their pharmacological activation ends ongoing swimming, whereas the inactivation of these neurons dramatically impairs the rapid termination of swimming. These neurons are henceforth referred to as stop cells, because they play a crucial role in the termination of locomotion. Our findings contribute to the fundamental understanding of motor control and provide important details about the cellular mechanisms involved in locomotor termination.

In vertebrates, stimulation of the mesencephalic locomotor region (MLR) on one side evokes symmetrical locomotor movements on both sides. How this occurs was previously examined in detail in a swimmer using body undulations (lamprey), but in tetrapods the downstream projections from the MLR to brainstem neurons are not fully understood. Here we examined the brainstem circuits from the MLR to identified reticulospinal neurons in the salamander Notophthalmus viridescens. Using neural tracing, we show that the MLR sends bilateral projections to the middle reticular nucleus (mRN, rostral hindbrain) and the inferior reticular nucleus (iRN, caudal hindbrain). Ca(2+) imaging coupled to electrophysiology in in vitro isolated brains revealed very similar responses in reticulospinal neurons on both sides to a unilateral MLR stimulation. As the strength of MLR stimulation was increased, the responses increased in size in reticulospinal neurons of the mRN and iRN, but the responses in the iRN were smaller. Bath-application or local microinjections of glutamatergic antagonists markedly reduced reticulospinal neuron responses, indicating that the MLR sends glutamatergic inputs to reticulospinal neurons. In addition, reticulospinal cells responded to glutamate microinjections and the size of the responses paralleled the amount of glutamate microinjected. Immunofluorescence coupled with anatomical tracing confirmed the presence of glutamatergic projections from the MLR to reticulospinal neurons. Overall, we show that the brainstem circuits activated by the MLR in the salamander are organized similarly to those previously described in lampreys, indicating that the anatomo-physiological features of the locomotor drive are well conserved in vertebrates. J. Comp. Neurol. 524:1361-1383, 2016. © 2015 The Authors The Journal of Comparative Neurology Published by Wiley Periodicals, Inc.

In 1966, Shik, Severin and Orlovskii discovered that electrical stimulation of a region at the junction between the midbrain and hindbrain elicited controlled walking and running in the cat. The region was named Mesencephalic Locomotor Region (MLR). Since then, this locomotor center was shown to control locomotion in various vertebrate species, including the lamprey, salamander, stingray, rat, guinea-pig, rabbit or monkey. In human subjects asked to imagine they are walking, there is an increased activity in brainstem nuclei corresponding to the MLR (i.e. pedunculopontine, cuneiform and subcuneiform nuclei). Clinicians are now stimulating (deep brain stimulation) structures considered to be part of the MLR to alleviate locomotor symptoms of patients with Parkinson's disease. However, the anatomical constituents of the MLR still remain a matter of debate, especially relative to the pedunculopontine, cuneiform and subcu- neiform nuclei. Furthermore, recent studies in lampreys have revealed that the MLR is more complex than a simple relay in a serial de- scending pathway activating the spinal locomotor circuits. It has multiple functions. Our goal is to review the current knowledge relative to the anatomical constituents of the MLR, and its physiological role, from lamprey to man. We will discuss these results in the context of the recent clinical studies involving stimulation of the MLR in patients with Parkinson's disease.

Skip to Next Section Olfactory sensory neurons innervate the olfactory bulb, where responses to different odorants generate a chemotopic map of increased neural activity within different bulbar regions. In this study, insight into the basal pattern of neural organization of the vertebrate olfactory bulb was gained by investigating the lamprey. Retrograde labelling established that lateral and dorsal bulbar territories receive the axons of sensory neurons broadly distributed in the main olfactory epithelium and that the medial region receives sensory neuron input only from neurons projecting from the accessory olfactory organ. The response duration for local field potential recordings was similar in the lateral and dorsal regions, and both were longer than medial responses. All three regions responded to amino acid odorants. The dorsal and medial regions, but not the lateral region, responded to steroids. These findings show evidence for olfactory streams in the sea lamprey olfactory bulb: the lateral region responds to amino acids from sensory input in the main olfactory epithelium, the dorsal region responds to steroids (taurocholic acid and pheromones) and to amino acids from sensory input in the main olfactory epithelium, and the medial bulbar region responds to amino acids and steroids stimulating the accessory olfactory organ. These findings indicate that olfactory subsystems are present at the base of vertebrate evolution and that regionality in the lamprey olfactory bulb has some aspects previously seen in other vertebrate species.

This review focuses on past and recent findings that have contributed to characterize the neural networks controlling respiration in the lamprey, a basal vertebrate. As in other vertebrates, respiration in lampreys is generated centrally in the brainstem. It is characterized by the presence of a fast and a slow respiratory rhythm. The anatomical and the basic physiological properties of the neural networks underlying the generation of the fast rhythm have been more thoroughly investigated; less is known about the generation of the slow respiratory rhythm. Comparative aspects with respiratory generators in other vertebrates as well as the mechanisms of modulation of respiration in association with locomotion are discussed.

Sensorimotor transformation is one of the most fundamental and ubiquitous functions of the central nervous system (CNS). Although the general organization of the locomotor neural circuitry is relatively well understood, less is known about its activation by sensory inputs and its modulation. Utilizing the lamprey model, a detailed understanding of sensorimotor integration in vertebrates is emerging. In this article, we explore how the vertebrate CNS integrates sensory signals to generate motor behavior by examining the pathways and neural mechanisms involved in the transformation of cutaneous and olfactory inputs into motor output in the lamprey. We then review how 5-hydroxytryptamine (5-HT) acts on these systems by modulating both sensory inputs and motor output. A comprehensive review of this fundamental topic should provide a useful framework in the fields of motor control, sensorimotor integration and neuromodulation.

Central networks modulate sensory transmission during motor behavior. Sensory inputs may thus have distinct impacts according to the state of activity of the central networks. Using an in-vitro isolated lamprey brainstem preparation, we investigated whether a brainstem locomotor center, the mesencephalic locomotor region (MLR), modulates sensory transmission. The synaptic responses of brainstem reticulospinal (RS) cells to electrical stimulation of the sensory trigeminal nerve were recorded before and after electrical stimulation of the MLR. The RS cell synaptic responses were significantly reduced by MLR stimulation and the reduction of the response increased with the stimulation intensity of the MLR. Bath perfusion of atropine prevented the depression of sensory transmission, indicating that muscarinic receptor activation is involved. Previous studies have shown that, upon stimulation of the MLR, behavioral activity switches from a resting state to an active-locomotor state. Therefore, our results suggest that a state-dependent modulation of sensory transmission to RS cells occurs in the behavioral context of locomotion and that muscarinic inputs from the MLR are involved.

The brainstem locomotor system is believed to be organized serially from the mesencephalic locomotor region (MLR) to reticulospinal neurons, which in turn project to locomotor neurons in the spinal cord. We identified brainstem muscarinoceptive neurons in lampreys (Petromyzon marinus) that received parallel inputs from the MLR and projected back to reticulospinal cells to amplify and extend the duration of locomotor output. These cells responded to muscarine with extended periods of excitation, received direct muscarinic excitation from the MLR and projected glutamatergic excitation to reticulospinal neurons. Targeted blockade of muscarine receptors over these neurons profoundly reduced MLR-induced excitation of reticulospinal neurons and markedly slowed MLR-evoked locomotion. The presence of these neurons forces us to rethink the organization of supraspinal locomotor control, to include a sustained feedforward loop that boosts locomotor output.

A unilateral activation of the mesencephalic locomotor region (MLR) produces symmetrical bilateral locomotion in all vertebrate species tested to date. How this occurs remains unresolved. This study examined the possibility that the symmetry occurred at the level of the inputs from the MLR to reticulospinal (RS) cells. In lamprey semi-intact preparations, we recorded intracellular responses of pairs of large, homologous RS cells on both sides to stimulation of the MLR on one side. The synaptic responses on both sides were very similar in shape, amplitude, and threshold intensity. Increasing MLR stimulation intensity produced a symmetrical increase in the magnitude of the responses on both sides. Ca2+imaging confirmed the bilateral activation of smaller-sized RS cells as well. In a high-divalent cation solution, the synaptic responses of homologous RS cells persisted and exhibited a constant latency during high-frequency stimulation. Moreover, during gradual replacement of normal Ringer's solution with a Ca2+-free solution, the magnitude of responses showed a gradual reduction with a similar time course in the homologous RS cells. These results support the idea that the MLR projects monosynaptically to RS cells on both sides with symmetrical inputs. During locomotion of the semi-intact preparation, the discharge pattern was also very similar in homologous bilateral RS cells. Anatomical experiments confirmed the presence of MLR neurons projecting ipsilaterally to the reticular formation intermingled with neurons projecting contralaterally. We conclude that the bilaterally symmetrical MLR inputs to RS cells are likely contributors to generating symmetrical locomotor activity.

Sensory inputs are subjected to modulation by central neural networks involved in controlling movements. It has been shown that serotonin (5-HT) modulates sensory transmission. This study examines in lampreys the effects of 5-HT on sensory transmission to brainstem reticulospinal (RS) neurons and the distribution of 5-HT cells that innervate RS cells. Cells were recorded intracellularly in the in vitro isolated brainstem of larval lampreys. Trigeminal nerve stimulation elicited disynaptic excitatory responses in RS neurons, and bath application of 5-HT reduced the response amplitude with maximum effect at 10 mum. Local ejection of 5-HT either onto the RS cells or onto the relay cells decreased sensory-evoked excitatory postsynaptic potentials (EPSPs) in RS cells. The monosynaptic EPSPs elicited from stimulation of the relay cells were also reduced by 5-HT. The reduction was maintained after blocking either N-methyl-d-aspartate (NMDA) or alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptors. The local ejection of glutamate over RS cells elicited excitatory responses that were only slightly depressed by 5-HT. In addition, 5-HT increased the threshold for eliciting sustained depolarizations in response to trigeminal nerve stimulation but did not prevent them. Combined 5-HT immunofluorescence with axonal tracing revealed that the 5-HT innervation of RS neurons of the middle rhombencephalic reticular nucleus comes mainly from neurons in the isthmic region, but also from neurons located in the pretectum and caudal rhombencephalon. Our results indicate that 5-HT modulates sensory transmission to lamprey brainstem RS cells.

The role of opioid receptors in modulating respiratory activity was investigated in in vitro brainstem preparations of adult lampreys by bath application of agonists and antagonists. The vagal motor output was used to monitor respiratory activity. Neuronal recordings were also performed to characterize the rostrolateral trigeminal region that has been suggested to be critical for respiratory rhythmogenesis. Microinjections of the micro-opioid receptor agonist [d-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin (DAMGO) were also made into this region and at different locations within the brainstem. Bath application of DAMGO (0.5-2 microM) caused marked decreases in respiratory frequency up to complete apnea. Bath application of the delta-opioid receptor agonist [d-Pen(2,5)]-enkephalin (DPDPE) at 10-40 microM induced less pronounced depressant respiratory effects, while no changes in respiratory activity were induced by the kappa-opioid receptor agonist trans-(1S,2S)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)cyclohexyl] benzeneacetamide (U50488) at 10-40 microM. Bath application of the opioid receptor antagonists naloxone and naltrindole did not affect baseline respiratory activity, but prevented agonist-induced effects. DAMGO microinjections (1 mM; 0.5-1 nl) at sites rostrolateral to the trigeminal motor nucleus, where respiration-related neuronal activity was recorded, abolished the respiratory rhythm. The results show that opioids may have an important role in the lamprey respiratory network and that micro-opioid receptor activation is the most effective in causing respiratory depression. They also indicate that endogenous opioids are not required for the generation of baseline respiratory activity. Apneic responses induced by DAMGO microinjections support the hypothesis that a specific opioid-sensitive region rostrolateral to the trigeminal motor nucleus, that we have termed the paratrigeminal respiratory group (pTRG), likely has a pivotal role in respiratory rhythmogenesis. Since the lamprey diverged from the main vertebrate line around 450 million years ago, our results also imply that the inhibitory role of opioids on respiration is present at an early stage of vertebrate evolution.

Brainstem networks generating the respiratory rhythm in lampreys are still not fully characterized. In this study, we described the patterns of respiratory activities and we identified the general location of underlying neural networks. In a semi-intact preparation including the brain and gills, rhythmic discharges were recorded bilaterally with surface electrodes placed over the vagal motoneurons. The main respiratory output driving rhythmic gill movements consisted of short bursts (40.9+/-15.6 ms) of discharge occurring at a frequency of 1.0+/-0.3 Hz. This fast pattern was interrupted by long bursts (506.3+/-174.6 ms) recurring with an average period of 37.4+/-24.9 s. After isolating the brainstem by cutting all cranial nerves, the frequency of the short respiratory bursts did not change significantly, but the slow pattern was less frequent. Local injections of a glutamate agonist (AMPA) and antagonists (6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) or D,L-amino-5-phosphonopentanoic acid (AP5)) were made over different brainstem regions to influence respiratory output. The results were similar in the semi-intact and isolated-brainstem preparations. Unilateral injection of AP5 or CNQX over a rostral rhombencephalic region, lateral to the rostral pole of the trigeminal motor nucleus, decreased the frequency of the fast respiratory rhythm bilaterally or stopped it altogether. Injection of AMPA at the same site increased the rate of the fast respiratory rhythm and decreased the frequency of the slow pattern. The activity recorded in this area was synchronous with that recorded over the vagal motoneurons. After a complete transverse lesion of the brainstem caudal to the trigeminal motor nucleus, the fast rhythm was confined to the rostral area, while only the slow activity persisted in the vagal motoneurons. Our results support the hypothesis that normal breathing depends on the activity of neurons located in the rostral rhombencephalon in lampreys, whereas the caudal rhombencephalon generates the slow pattern.

In vertebrates, locomotion is associated with changes in respiratory activity, but the neural mechanisms by which this occurs remain unknown. We began examining this in lampreys using a semi-intact preparation of young adult Petromyzon marinus, in which respiratory and locomotor behaviors can be recorded simultaneously with the activity of the underlying neural control systems. Spontaneous fictive respiration was recorded with suction electrodes positioned over the glossopharyngeal or the rostral vagal motor nucleus. In this preparation, locomotor activity, characterized by symmetrical tail movements (electromyogram recordings), was evoked by mechanical stimulation of the skin. During locomotion, the mean respiratory frequency and the mean area of the motor bursts were significantly increased (81.6+/-28.6% and 62.8+/-25.4%, respectively; P0.05). The frequency returned to normal 92+/-51 s after the end of locomotion. There were fluctuations in the instantaneous respiratory and locomotor frequencies that were rhythmical but antiphasic for the two rhythmic activities. The changes in respiratory activity were also examined during bouts of locomotion occurring spontaneously, and it was found that a modification in respiratory activity preceded the onset of spontaneous locomotion by 3.5+/-2.6 s. This suggests that the early respiratory changes are anticipatory and are not caused by feedback generated by locomotion. The increase in respiratory frequency during locomotion induced by sensory stimulation persisted after removal of the mesencephalon. When both the mesencephalon and spinal cord were removed, resulting in the isolation of the rhombencephalon, changes in the respiratory activity were also present following skin stimulations that would have normally induced locomotion. Altogether, the results suggest that respiratory changes are programmed to adjust ventilation prior to motor activity, and that a central rhombencephalic mechanism is involved.

The localization of gamma-aminobutyric acid (GABA) has been well described in most classes of vertebrates but not in adult lampreys. The question if the GABA distribution is similar throughout the vertebrate subphylum is therefore still to be addressed. We here investigate two lamprey species, the sea lamprey, Petromyzon marinus, and the river lamprey, Lampetra fluviatilis, and compare the GABA pattern with that of other vertebrates. The present immunohistochemical study provides an anatomical basis for the general distribution and precise localization of GABAergic neurons in the adult lamprey forebrain and brainstem. GABA-immunoreactive cells were organized in a virtually identical manner in the two species. They were found throughout the brain, with the following regions being of particular interest: the granular cell layer of the olfactory bulb, the nucleus of the anterior commissure, the septum, the lateral and medial pallia, the striatum, the nucleus of the postoptic commissure, the thalamus, the hypothalamus, and pretectal areas, the optic tectum, the torus semicircularis, the mesencephalic tegmentum, restricted regions of the rhombencephalic tegmentum, the octavolateral area, and the dorsal column nucleus. The GABA distribution found in cyclostomes is very similar to that of other classes of vertebrates, including mammals. Since the lamprey diverged from the main vertebrate line around 450 million years ago, this implies that already at that time the basic vertebrate plan for the GABA innervation in different parts of the brain had been developed.

This study examined the spatial and temporal distribution of serotonin-immunoreactive (5-HT-ir) neurons in the brainstem of Petromyzon marinus at three developmental stages, larval, postmetamorphic, and reproductive. Computer-assisted 3-D reconstructions were made of the three main 5-HT-ir neuron groups. The rostralmost brainstem group was located near the posterior commissure, the second group at the isthmus, and the third group in the bulbar area. For each of those groups, the distribution of the 5-HT-ir neurons was very similar in the three developmental stages examined, suggesting that the 5-HT system is relatively mature early in larval animals. The soma of 5-HT-ir neurons increased in size and their dendritic fields increased in complexity with development. Furthermore, the number of 5-HT-ir neurons in each group increased significantly from the larval to the reproductive stage. To determine whether this was due to the genesis of 5-HT neurons, bromodeoxyuridine (BrdU) was injected into larval, metamorphosing, and postmetamorphic lampreys. These experiments revealed a few neurons colocalizing BrdU and 5-HT in metamorphosing animals. Taken together, the present results suggest that 5-HT neurons increase in number during maturation and that neurogenesis could, at least partially, contribute to the appearance of new 5-HT cells at different developmental stages. J. Comp. Neurol. 495:788–800, 2006. © 2006 Wiley-Liss, Inc.

Substance P initiates locomotion when injected in the brain stem of mammals. This study examined the possible role of this peptide on the supraspinal locomotor command system in lampreys. Substance P was bath applied or locally injected into an in vitro isolated brain stem, and the effects of the drug were examined on reticulospinal cells and on the occurrence of swimming in a semi-intact preparation. Bath applications of substance P induced sustained depolarizations occurring rhythmically in intracellularly recorded reticulospinal cells. Spiking activity was superimposed on the depolarizations and swimming was induced. The sustained depolarizations were abolished by tetrodotoxin, and substance P did not affect the membrane resistance of reticulospinal cells nor their firing properties, suggesting that it did not directly effect reticulospinal cells. To establish where the effects were exerted, successive lesions of the brain stem were made as well as local applications of the drug in the brain stem. Removing the mesencephalon abolished the sustained depolarizations, whereas large ejections of the drug in the mesencephalon excited reticulospinal cells and elicited bouts of swimming. More local injections into the mesencephalic locomotor region (MLR) also elicited swimming. After an injection of substance P, the current threshold needed to induce locomotion by MLR stimulation was decreased, and the size of the postsynaptic responses of reticulospinal cells to MLR stimulation was increased. Substance P also reduced the frequency of miniature spontaneous postsynaptic currents in reticulospinal cells. Taken together, these results suggest that substance P plays a neuromodulatory role on the brain stem locomotor networks of lampreys.

This study was carried out to identify lamprey neurones relaying trigeminal sensory inputs to reticulospinal cells. Double labeling with fluorescent tracers was used in vitro. Fluorescein-conjugated dextran amines were applied to the proximal stump of the cut trigeminal nerve on both sides, and Texas Red-conjugated dextran amines were injected unilaterally in the middle (MRRN) or the posterior (PRRN) rhombencephalic reticular nuclei. Texas Red retrogradely labeled cells were found ipsi- and contralateral to each injection. Any of these cells with the soma or at least a major dendrite among the fluorescein-labeled trigeminal afferent axons was considered a candidate relay cell. Of these two possibilities, only cells with their soma among the fluorescein-labeled trigeminal afferents were found. The candidate relay cells projecting to the MRRN were mostly clustered at the caudal vestibular nerve level within the trigeminal descending tract, whereas the majority of those projecting to the PRRN were located more caudally. The diameter of candidate relay cells ranged from 9.2 to 24.6 mum and 9.2 to 46.1 mum, after MRRN and PRRN injections, respectively. A possible relay function for these cells was tested with electrophysiological experiments. The intracellular responses to trigeminal nerve stimulation were recorded in reticulospinal cells under control conditions and after ejections of a combination of glutamate ionotropic receptor antagonists over the candidate relay cells in small areas along the sulcus limitans. The synaptic responses elicited in MRRN reticulospinal cells were maximally depressed when ejections were made at the level of the vestibular nerve, in accord with the anatomical data. The synaptic responses in PRRN reticulospinal cells showed maximal depression when ejections were made slightly more caudally. Altogether, these results suggest that cells located within the trigeminal descending tract and projecting to reticular nuclei are likely to be the sensory trigeminal relays to reticulospinal neurones in lampreys.

In lampreys as in other vertebrates, brainstem centres play a key role in the initiation and control of locomotion. One such centre, the mesencephalic locomotor region (MLR), was identified physiologically at the mesopontine border. Descending inputs from the MLR are relayed by reticulospinal neurons in the pons and medulla, but the mechanisms by which this is carried out remain unknown. Because previous studies in higher vertebrates and lampreys described cholinergic cells within the MLR region, we investigated the putative role of cholinergic agonists in the MLR-controlled locomotion. The local application of either acetylcholine or nicotine exerted a direct dose-dependent excitation on reticulospinal neurons as well as induced active or fictive locomotion. It also accelerated ongoing fictive locomotion. Choline acetyltransferase-immunoreactive cells were found in the region identified as the MLR of lampreys and nicotinic antagonists depressed, whereas physostigmine enhanced the compound EPSP evoked in reticulospinal neurons by electrical stimulation of this region. In addition, cholinergic inputs from the MLR to reticulospinal neurons were found to be monosynaptic. When the brainstem was perfused with d-tubocurarine, the induction of swimming by MLR stimulation was depressed, but not prevented, in a semi-intact preparation. Altogether, the results support the hypothesis that cholinergic inputs from the MLR to reticulospinal cells play a substantial role in the initiation and the control of locomotion.

The distribution of vestibular primary afferents as well as their relationship with vestibulospinal and other brainstem neurons were studied in lampreys using anatomical tracers. Afferents from the anterior (aVIIIn) and the posterior (pVIIIn) branches of the vestibular nerve were located mainly in the ventral nucleus of the octavolateral area. The relationship between afferents and vestibulospinal neurons was studied by applying one fluorescent tracer to the whole vestibular nerve or one of its branches and applying another tracer to the spinal cord. Some afferents showed large, bulb-like enlargements (bulbs) and about 20 of these were found in the anterior and the intermediate octavomotor nucleus, whereas about 40 were found in the posterior octavomotor nucleus. Some of the bulbs made apparent contact with vestibulospinal neurons in the intermediate octavomotor nucleus and originated mostly from the aVIIIn, whereas bulbs in the posterior octavomotor nucleus originated from the pVIIIn. Applications of biocytin to hemisegments of rostral spinal cord labeled vestibulospinal neurons located in the ipsilateral intermediate octavomotor nucleus and the contralateral posterior octavomotor nucleus. In addition, vestibular primary afferents with bulbs in apparent contact with vestibulospinal neurons were transneuronally labeled by biocytin. They were observed in the ipsilateral aVIIIn and the contralateral pVIIIn and could be followed in the labyrinths, where they innervated the vertical and horizontal arms of the semicircular canal crests. Taken together, these results indicate that vestibular primary afferents from the aVIIIn innervate predominantly vestibulospinal neurons of the intermediate octavomotor nucleus, whereas afferents from the pVIIIn innervate vestibulospinal neurons in the posterior octavomotor nucleus. This anatomical organization suggests that afferents carrying bulbs convey dynamic information to vestibulospinal neurons, which, in turn, project to the spinal cord networks.

The present study was carried out to characterize anatomically the vestibulospinal (VS) system of lampreys. Cobalt-lysine or Texas Red dextran amines were applied in vitro to the rostral spinal cord. Two distinct populations of VS neurons were labeled in the ventral nucleus of the area octavolateralis. The rostral group, comprising the intermediate octavomotor nucleus (ION), contained between 100 and 150 neurons, having somata of variable size and morphology. Intracellular injections of Lucifer Yellow in single neurons revealed ION VS neurons with dendrites extending in the ventrolateral alar plate as well as medially in the basal plate. The caudal group, comprising the posterior octavomotor nucleus (PON), contained approximately 65 neurons, most of which were unipolar with round or oval somata. To study the projections of VS axons, cobalt-lysine was injected into the ION or PON regions in the brainstem. Axons from the ION projected to the ipsilateral spinal cord, whereas PON axons decussated within the basal plate giving out descending and ascending branches. The descending branch projected to the contralateral spinal cord. Injections of two fluorescent dextran-amines, each restricted to one side of the spinal cord, did not double-label VS cells in either octavomotor nuclei, indicating that the projections of each nucleus are restricted to one side. Injections of horseradish peroxidase further caudally in the spinal cord revealed that VS axons from the ION reached past the gill region. Our results indicate that the organization of the VS system of lampreys is similar to that observed in other vertebrates. J. Comp. Neurol. 407:512‐526, 1999. r 1999 Wiley-Liss, Inc.

The cell bodies of efferent neurons supplying the masseter and digastric muscles of the rabbit are located in two brainstem nuclei: the trigeminal motor nucleus and cell group k. The latter also contains neurons innervating muscles of the middle ear and Eustachian tube, as well as neurons that project to the cerebellum and the oculomotor complex. As part of an attempt to identify the functional subpopulations within the three cell divisions (k1–k3) that make up cell group k, we have investigated the distribution of neurons containing choline acetyltransferase, because these are likely to be motoneurons. Five rabbits anaesthetized with sodium pentobarbital (90 mg/kg, i.v.) were used in this study. They were perfused with 4% paraformaldehyde and 0.1% glutaraldehyde in phosphate buffer (0.1 M, pH 7.4). Two animals were used for preliminary studies. In the other three cases, serial Vibratome coronal sections of the brainstem were cut at 50 μ m and two series of alternating sections were collected. The first was stained with a monoclonal antibody (code AB8, Incstar) directed against choline acetyltransferase, using the avidin–biotin–peroxidase method. The other was stained with Cresyl Violet. Cell counts and three-dimensional reconstructions were made for both series to determine positions and ratios of cholinergic and non-cholinergic neurons within the trigeminal motor nucleus and the subdivisions of cell group k. The results showed that the numbers of choline acetyltransferase- and Nissl-stained neurons within the trigeminal motor nucleus were almost identical. In cell group k, significantly fewer choline acetyltransferase-stained cells were counted in all three animals (ratios of choline acetyltransferase/Nissl=0.53–0.71). In addition, the distribution of cholinergic neurons was not uniform throughout cell group k. Subdivisions k1 and k3 contained proportionately less choline acetyltransferase-positive cells (ratios of choline acetyltransferase/Nissl=0.23–0.64) than did k2 (ratios choline acetyltransferase/Nissl=0.75–0.88). Within each subdivision, there were significant differences in the spatial coordinates of Nissl- and choline acetyltransferase-positive neurons. We conclude that cell group k contains at least two populations of neurons which are unevenly distributed between and within the three subdivisions. While the majority of neurons in subgroup k2 contain choline acetyltransferase and thus are likely to be motoneurons, more than half of the neurons in subgroups k1 and k3 are not cholinergic. It remains to be determined whether these are the neurons that project to the cerebellum and to other CNS regions.

Behavioral studies in lampreys of the northern genera, Ichthyomyzon, reveal that sensory inputs initiate and modulate locomotion by activation of reticulospinal (RS) neurones, which constitute the primary descending system involved in motor activity. The interneurones relaying afferent vestibular, trigeminal, lateral line, cutaneous and proprioceptive inputs are localized in the rhombencephalic region of the lamprey brainstem, unlike the visual inputs that are relayed in the mesencephalic region. The knowledge of diencephalic-mesencephalic cell distributions that project to the RS neurones is limited. They were isolated by iontophoretically injecting cobalt-lysine in vitro into the middle (MRRN) and posterior (PRRN) rhombencephalic reticular nuclei of Petromyzon marinus and Ichthyomyzon unicuspis, Fourteen of 31 injections were successful (MRRN, 7; PRRN, 7). Cell groups were labeled ipsilateral to the injection site in the thalamus (corpi geniculati; pars dorsalis thalami lateralis and medialis; nucleus (n.) subhabenularis lateralis), in the epithalamus (n. commissura posteriori) and in the pretectum. Cell groups were labeled bilaterally within the dorsal region along the diencephalic-mesencephalic border (caudal pretectum and rostral tectum opticum), in tectum opticum, torus semicircularis, and tegmentum mesencephali. There were more backfilled cells from MRRN injections (538-6466 cells) than from PRRN injections (53-553 cells) (MW Rank Sum, p < 0.001). The cell bodies were less than 40 microns long ipsilateral to the injection site, and longer contralaterally. Those greater than 50 microns were backfilled from PRRN injections. The location and organization of the cell groups identified is comparable to that of other vertebrates.

Tectoreticular (TR) cells along the diencephalic-mesencephalic border are the origin of prominent crossed and uncrossed pathways that project to the middle (MRRN) and posterior (PRRN) rhombencephalic reticular nuclei in juvenile and adult lampreys [I.C. Zompa, R. Dubuc, Diencephalic and mesencephalic projections to rhombencephalic reticular nuclei in lampreys, Brain Res. (1998) in press.]. This study investigated the synaptic contacts between TR axons and the reticular cells. Intracellular recordings were carried out in reticular neurones (n=124) while microstimulating the TR regions. Tectoreticular inputs were recorded in all reticular cells studied (248 PSPs); although stronger responses were evoked in the MRRN neurones. The majority of responses were excitatory, but increasingly mixed and inhibitory when recorded in the middle and caudal part of the reticular nuclei. The excitation had the shortest onset latencies and sharpest slopes measured in both reticular nuclei, while the inhibition was longer and smoother. The characteristics of TR inputs to different reticular cell types is also presented. The transmission of evoked responses was isolated to the crossed and uncrossed TR pathways by studying the effects of 1% Xylocaine ejections and surgical lesions. The TR inputs were transmitted to reticular cells through monosynaptic and polysynaptic contacts. The synaptic transmission involved excitatory amino acids, acting through AMPA and NMDA receptors, while the inhibition was glycinergic. Comparisons with other sensory systems in lampreys are discussed.

The present study was aimed at identifying spinal neurons ascending to the brainstem outside the dorsal columns in the lamprey. Two retrograde tracers (cobalt-lysine and horseradish peroxidase [HRP]) were injected in the brainstem or rostral spinal cord in vivo or in vitro. Labeled cells were distributed bilaterally with a contralateral dominance, along the whole rostrocaudal extent of the spinal cord. The density of cells markedly decreased rostrocaudally. Several classes of brainstem-projecting neurons were identified. Most cells with a short axon were small and formed columns, in the dorsolateral and ventrolateral gray matter, at the transition between the rhombencephalon and the spinal cord. Dorsal elongated cells were spindle shaped, located medially, in the first two spinal segments. Lateral elongated cells were medium to large size neurons, located in the intermediate and lateral gray matter, mainly contralateral to the injection site. Their axon emerging from the lateral part of the soma crossed the midline, ventral to the central canal. These cells were present throughout the rostral spinal cord. Cells were also labeled in the lateral white matter. Some of them had the typical dendritic arborizations of edge cells (intraspinal stretch receptor neurons) and were located in the most rostral segments, bilaterally. Other medium to large size neurons were identified dorsal and medial to most of the edge cells. We suggest that at least the group of lateral elongated cells exhibits rhythmic membrane potential oscillations during fictive locomotion. These cells may, together with the rostral edge cells, be responsible for the locomotor-related modulation of activity in reticulospinal and vestibulospinal neurons.

Previous studies have shown that the masseter muscle is supplied by motoneurons located in the anterodorsal region of the trigeminal motor nucleus and by an additional group of efferent neurons located in cell group k. The present experiments were performed on nine rabbits and were designed to establish the locations of neurons innervating the different regions of this muscle. Retrograde labeling with two fluorescent tracers (FluoroGold and Fast Blue) was applied to the central ends of cut branches of the masseter nerve. Serial coronal sections of the brainstem were viewed with fluorescence microscopy. The labeled cells were counted in all animals, and three-dimensional reconstructions of their distribution were made in five cases. In each successful experiment, labeled neurons were seen in the anterodorsal region of the trigeminal motor nucleus and in the two dorsal cell columns of cell group k (k1 and k3). Within-animal comparisons of the median position of populations innervating two distinct muscle regions in five rabbits showed that there were no significant differences in either the dorsoventral or rostrocaudal axes. However, in each case, there was a small but significant difference (83-173 microm) in the mediolateral axis within the motor nucleus but not within cell group k. Even in this axis, there was a 94-99% overlap of the two populations. Comparisons of the neuronal cross-sectional area showed that the deep regions were innervated by a larger proportion of small neurons from both nuclei than were the superficial and intermediate regions. Our results suggest that there is no simple topographical arrangement of motoneurons that corresponds to the peripheral pattern of nerve supply to the different regions of the masseter muscle.

The effects of stimulating the reticular formation were studied during fictive locomotion in lampreys ( Ichthyomyzon unicuspis ). The in vitro isolated preparation of the brainstem and spinal cord was used and fictive locomotion was induced by bath application of N -methyl- d -aspartate (NMDA; 50–100 μM). During different phases of the locomotor cycle, short trains of stimuli (10 pulses at 80–100 Hz; 10 μA) were delivered through glass-coated tungsten microelectrodes positioned within the middle rhombencephalic reticular nucleus (MRRN) and their effects were studied on ipsi- and contralateral ventral root locomotor discharges. Irrespective of the locomotor phase during which the stimulation train was delivered, a resetting effect occurred. It was characterized by a re-synchronization of the locomotor discharges with a constant latency for each ventral root on the ipsilateral side. The latency increased as the recorded root was located further caudally. This increase in latency was in the range of the phase lag observed between roots during control bouts of locomotion. These results suggest that reticulospinal neurones exert strong resetting effects on spinal locomotor networks. These effects may play a significant role with respect to changes of direction during swimming.

Although there is abundant evidence for segregated processing in the olfactory system across vertebrate taxa, the spatial relationship between the second order projection neurons (PNs) of olfactory subsystems connecting sensory input to higher brain structures is less clear. In the sea lamprey, there is tight coupling between olfaction and locomotion via PNs extending to the posterior tuberculum from the medial region of the olfactory bulb. This medial region receives peripheral input predominantly from the accessory olfactory organ. However, the axons from olfactory sensory neurons residing in the main olfactory epithelium extend to non-medial regions of the olfactory bulb, and the non-medial bulbar PNs extend their axons to the lateral pallium. It is not known if the receptive fields of the PNs in the two output pathways overlap; nor has the morphology of these PNs been investigated. In this study, retrograde labelling was utilized to investigate the PNs belonging to medial and non-medial projections. The dendrites and somata of the medial PNs were confined to medial glomerular neuropil, and dendrites of non-medial PNs did not enter this territory. The cell bodies and dendrites of the non-medial PNs were predominantly located below the glomeruli (frequently deeper in the olfactory bulb). While PNs in both locations contained single or multiple primary dendrites, the somal size was greater for medial than for non-medial PNs. When considered with the evidence-to-date, this study shows different neuroanatomical organization for medial olfactory bulb PNs extending to locomotor control centers and non-medial PNs extending to the lateral pallium in this vertebrate.

Plasticity of synaptic transmission has been investigated in the lamprey brainstem. In this preparation, neurons of the vestibular nuclei make monosynaptic connections to reticulospinal neurons of the posterior (PRRN) and middle (MRRN) rhombencephalic reticular nuclei. This glutamatergic projection shows a long-lasting enhancement in efficacy following a brief high frequency (50 Hz) train of stimuli (tetanus). Enhancement was found on the inputs from the octavomotorius intermediate (nOMI) and octavomotorius posterior (nOMP) nuclei to the reticulospinal neurons of the PRRN and MRRN, and persisted for the entire duration of the recordings. The enhancement was limited to the pathway that received tetanic stimulation for the inputs to any given reticulospinal neuron. It depended upon the activation of postsynaptic processes at least in part; dialysis of the reticulospinal neuron by recording with a whole-cell patch pipette prevented its induction. This dialysis-dependent abolition of enhancement was shown to be dependent on a change in Ca2+ concentration in the postsynaptic neuron. The enhancement was not affected by blockade of NMDA receptors with D,L-2- amino-5-phosphonopentanoate (AP5) but was prevented by the metabotropic glutamate receptor (mGluR) antagonist (R,S)-alpha-methyl-4- carboxyphenylglycine (MCPG). In conclusion, this study demonstrates that vestibular inputs to reticulospinal neurons are capable of undergoing long-term potentiation (LTP) and that this LTP shows synapse specificity. Furthermore, this LTP is activated by synaptic stimulation of a subtype of the m-GluR family and its induction is mediated by changing Ca2+ concentrations in the postsynaptic neuron.

Chemical sensory signals play a crucial role in eliciting motor behaviors. We now review the different motor behaviors induced by chemosensory stimuli in fish as well as their neural substrate. A great deal of research has focused on migratory, reproductive, foraging, and escape behaviors but it is only recently that the molecules mediating these chemotactic responses have become well-characterized. Chemotactic responses are mediated by three sensory systems: olfactory, gustatory, and diffuse chemosensory. The olfactory sensory neuron responses to chemicals are now better understood. In addition, the olfactory projections to the central nervous system were recently shown to display an odotopic organization in the forebrain. Moreover, a specific downward projection underlying motor responses to olfactory inputs was recently described.

Recent research in the field of spinal pattern generation has concentrated on three main areas: the effects of various transmitters on spinal rhythmic patterns in reduced preparations (neonatal rats, chick embryos, tadpole embryos, lampreys); the changes in membrane properties of different elements of the generating circuits; and the interactions between central generating mechanisms and afferent inputs. The important message is that new properties of neural membranes, as well as new reflex responses, have been identified that could not have been predicted in the absence of such rhythmic activity.

In order to determine if reticulospinal neurons involved in the control of locomotion and responsive to exogenously applied 5-hydroxtryptamine (5-HT) are innervated by fibers that contain serotonin, the serotoninergic innervation of reticulospinal neurons, identified by retrograde labeling with fluorescein-conjugated dextran-amine (FDA), was investigated by immunohistochemistry in the lamprey brainstem. A widespread distribution of 5-HT immunoreactive (5-HT-ir) fibers was seen within the basal plate of the brainstem, an area containing reticulospinal somata and dendritic arborizations. Numerous 5-HT varicose fibers were found in close relation to large reticulospinal cell bodies, particularly in the middle and anterior rhombencephalic reticular nuclei (MRRN and ARRN). Some of these reticulospinal somata were surrounded by a very dense pericellular 5-HT innervation. 5-HT-ir fibers were also seen in other brain structures that are known to influence reticulospinal neurons such as the rhombencephalic alar plate containing sensory relay interneurons, cranial nerves (III–X), cerebellum, and tectum. These findings suggest that, as in the spinal cord, motor behavior controlled by reticulospinal neurons may be subject to a serotoninergic modulation. © 1994 Wiley-Liss, Inc.

This study examines the connectivity in the neural networks controlling respiration in the lampreys, a basal vertebrate. Previous studies have shown that the lamprey paratrigeminal respiratory group (pTRG) plays a crucial role in the generation of respiration. By using a combination of anatomical and physiological techniques, we characterized the bilateral connections between the pTRGs and descending projections to the motoneurons. Tracers were injected in the respiratory motoneuron pools to identify pre-motor respiratory interneurons. Retrogradely labeled cell bodies were found in the pTRG on both sides. Whole-cell recordings of the retrogradely labeled pTRG neurons showed rhythmical excitatory currents in tune with respiratory motoneuron activity. This confirmed that they were related to respiration. Intracellular labeling of individual pTRG neurons revealed axonal branches to the contralateral pTRG and bilateral projections to the respiratory motoneuronal columns. Stimulation of the pTRG induced excitatory postsynaptic potentials in ipsi- and contralateral respiratory motoneurons as well as in contralateral pTRG neurons. A lidocaine HCl (Xylocaine) injection on the midline at the rostrocaudal level of the pTRG diminished the contralateral motoneuronal EPSPs as well as a local injection of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and (2R)-amino-5-phosphonovaleric acid (AP-5) on the recorded respiratory motoneuron. Our data show that neurons in the pTRG send two sets of axonal projections: one to the contralateral pTRG and another to activate respiratory motoneurons on both sides through glutamatergic synapses. J. Comp. Neurol. 520:1442–1456, 2012. © 2011 Wiley Periodicals, Inc.

Locomotion is a basic motor function generated and controlled by genetically defined neuronal networks. The pattern of muscle synergies is generated in the spinal cord, whereas neural centers located above the spinal cord in the brainstem and the forebrain are essential for initiating and controlling locomotor movements. One such locomotor control center in the brainstem is the mesencephalic locomotor region (MLR), first discovered in cats and later found in all vertebrate species tested to date. Over the last years, we have investigated the cellular mechanisms by which this locomotor region operates in lampreys. The lamprey MLR is a well-circumscribed region located at the junction between the midbrain and hindbrain. Stimulation of the MLR induces locomotion with an intensity that increases with the stimulation strength. Glutamatergic and cholinergic monosynaptic inputs from the MLR are responsible for excitation of reticulospinal (RS) cells that in turn activate the spinal locomotor networks. The inputs are larger in the rostral than in the caudal hindbrain RS cells. MLR stimulation on one side elicits symmetrical excitatory inputs in RS cells on both sides, and this is linked to bilateral projections of the MLR to RS cells. In addition to its inputs to RS cells, the MLR activates a well-defined group of muscarinoceptive cells in the brainstem that feeds back strong excitation to RS cells in order to amplify the locomotor output. Finally, the MLR gates sensory inputs to the brainstem through a muscarinic mechanism. It appears therefore that the MLR not only controls locomotor activity but also filters sensory influx during locomotion.

Locomotion is a basic motor function generated and controlled by genetically defined neuronal networks. The pattern of muscle synergies is generated in the spinal cord, whereas neural centers located above the spinal cord in the brainstem and the forebrain are essential for initiating and controlling locomotor movements. One such locomotor control center in the brainstem is the mesencephalic locomotor region (MLR), first discovered in cats and later found in all vertebrate species tested to date. Over the last years, we have investigated the cellular mechanisms by which this locomotor region operates in lampreys. The lamprey MLR is a well-circumscribed region located at the junction between the midbrain and hindbrain. Stimulation of the MLR induces locomotion with an intensity that increases with the stimulation strength. Glutamatergic and cholinergic monosynaptic inputs from the MLR are responsible for excitation of reticulospinal (RS) cells that in turn activate the spinal locomotor networks. The inputs are larger in the rostral than in the caudal hindbrain RS cells. MLR stimulation on one side elicits symmetrical excitatory inputs in RS cells on both sides, and this is linked to bilateral projections of the MLR to RS cells. In addition to its inputs to RS cells, the MLR activates a well-defined group of muscarinoceptive cells in the brainstem that feeds back strong excitation to RS cells in order to amplify the locomotor output. Finally, the MLR gates sensory inputs to the brainstem through a muscarinic mechanism. It appears therefore that the MLR not only controls locomotor activity but also filters sensory influx during locomotion.