Biomedical subjects

R Dubuc

Publications and source records attributed to R Dubuc.

At least 19 recordsLinked to original sources

PubMed1997

Anatomical organization of efferent neurons innervating various regions of the rabbit masseter muscle.

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.

Animals↗

PubMed1997

Effects of stimulating the reticular formation during fictive locomotion in lampreys.

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 microM). During different phases of the locomotor cycle, short trains of stimuli (10 pulses at 80-100 Hz; 10 microA) 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.

Animals↗

PubMed1996

A mesencephalic relay for visual inputs to reticulospinal neurones in lampreys.

Visual stimuli elicit motor responses in lampreys. These responses rely, in part, on the activation of reticulospinal (RS) neurones which constitute the main descending pathway in these early vertebrates. This study sought to identify and characterize possible mesencephalic relays for visual inputs to RS neurones of the rhombencephalon. The anatomical substrate subserving this function was investigated by iontophoretically ejecting cobalt-lysine, a retrograde tracer, in the middle rhombencephalic reticular nucleus in the in vitro isolated brainstem preparation of young adult Petromyzon marinus. Several populations of cells were retrogradely labeled in the brainstem. Of particular interest were the cell populations found on each side of rostral mesencephalon, located in the tectum and pretectum. There were, on average, 113 cells labeled contralateral to the injection site and 80 cells labeled ipsilateral to the injection site. The cells were morphologically similar on both sides, except that the contralateral group had larger cell bodies as compared to the group on the ipsilateral side. To determine whether the axons of the cells contacted reticulospinal neurones, electrophysiological experiments were carried out in which the region containing these cells was microstimulated. Large post-synaptic potentials were recorded intracellularly in RS neurones. Furthermore, microstimulation of the optic nerve on the same side as the recorded cell (i ON) evoked responses with a pattern similar to those resulting from stimulation of the optic tectum contralateral to the cell recorded (co OT), except for the longer response latencies. Local ejection of xylocaine (1% lidocaine hydrochloride) or CNQX (1 mM) onto the co OT reversibly abolished the responses evoked from stimulation of the i ON. There were no significant effects observed when the drug was ejected onto optic tectum ipsilateral to the cell. Taken together, the results from this study indicate that the crossed tectoreticular pathway is involved in relaying optic nerve inputs to RS neurones of the middle rhombencephalic reticular nucleus. Moreover, cells of origin of this pathway appear, in all respect, homologous to cells giving rise to the crossed tectobulbar pathway in other vertebrates.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1995

Trigeminal inputs to reticulospinal neurones in lampreys are mediated by excitatory and inhibitory amino acids.

Reticulospinal (RS) neurones integrate sensory inputs from several modalities to generate appropriate motor commands for maintaining body orientation and initiation of locomotion in lampreys. As in other vertebrates, trigeminal afferents convey sensory inputs from the head region. The in vitro brainstem/spinal cord preparation of the lamprey was used for characterizing trigeminal inputs to RS neurones as well as the transmitter systems involved. The trigeminal nerve on each side was electrically stimulated and synaptic responses, which consisted of mixed excitation and inhibition, were recorded intracellularly in the middle and posterior rhombencephalic reticular nuclei. The EPSPs were mediated by activation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate receptors. An increase in the late phase of the excitatory response occurred when Mg2+ ions were removed from the Ringer's solution. This effect was antagonized by 2-amino-5-phosphonopentanoate (2-AP5) or reversed by restoring Mg2+ ions to the perfusate suggesting the activation of N-methyl-D-aspartate (NMDA) receptors. IPSPs were mediated by glycine. These findings are similar to those reported for other types of sensory inputs conveyed to RS neurones, where excitatory and inhibitory amino acid transmission is also involved.

Animals↗

PubMed1995

Long-term potentiation of glutamatergic pathways in the lamprey brainstem.

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.

Animals↗

PubMed1994

5-HT innervation of reticulospinal neurons and other brainstem structures in lamprey.

In order to determine if reticulospinal neurons involved in the control of locomotion and responsive to exogenously applied 5-hydroxytryptamine (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 aborizations. 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.

Animals↗

PubMed1994

Spinal pattern generation.

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.

Afferent Pathways↗

PubMed1994

Importance of primer selection in the application of PCR technology to the diagnosis of bovine leukemia virus.

The polymerase chain reaction (PCR) was used to detect bovine leukemia virus in bovine blood samples. When applied to leucocytes extracted from the blood samples, the standard method of DNA extraction gave good correlation with agar gel immunodiffusion, but a method in which 5 microliters of blood was the starting material was unreliable. Selection of the primers was important, and differences in results were observed when the PCR method was applied to blood samples from different geographic areas. The sensitivity varied from 50% to 90%, depending on the primer set applied to the gag gene of proviral nucleic acid. This variation was based on geographic origin of the cattle, suggesting an influence of viral strain. In some areas, more than 1 primer may needed to optimize results.

Animals↗

PubMed1993

Glutamate metabotropic receptor mediated depression of synaptic inputs to lamprey reticulospinal neurones.

The transmission of vestibular inputs to reticulospinal (RS) neurones of the posterior rhombencephalic nucleus (PRRN) has been shown to be depressed by the bath application of N-methyl-D-aspartate (NMDA). The aim of this study was to investigate the pharmacological mechanism involved using patch clamp recordings of reticulospinal neurones. It is demonstrated that the chemical component of vestibular inputs to the PRRN is mediated by glutamatergic synapses utilising alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-D-aspartate (NMDA) receptors on the PRRN neurones. Monosynaptic excitatory postsynaptic currents (EPSCs) from octavomotorius relay cells to RS neurones are markedly depressed by the application of NMDA, a depression which was insensitive to competitive and non-competitive NMDA receptor antagonists. The effect of NMDA was eliminated by inactivation of G proteins. A similar depressive effect was observed following application of (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD) to the superfusate. It is concluded that NMDA acts at a metabotropic receptor located most likely presynaptically to reticulospinal neurones on terminals of octavomotorius relay cells.

2-Amino-5-phosphonovalerate↗

PubMed1993

Dorsal root and dorsal column mediated synaptic inputs to reticulospinal neurons in lampreys: involvement of glutamatergic, glycinergic, and GABAergic transmission.

This study was aimed at characterizing the inputs from dorsal roots and dorsal columns to reticulospinal neurons within the posterior rhombencephalic reticular nucleus in the lamprey. The in vitro isolated brainstem and spinal cord preparation was used. Microstimulation of dorsal roots and columns on both sides induced, in identified reticulospinal neurons, synaptic responses which consisted of large IPSPs mixed with excitation, particularly from stimulation on the ipsilateral side. When the spinal cord was selectively exposed to kynurenic acid or to Ca2+ free Ringer's containing 2mM Mn2+, synaptic responses to stimulation of dorsal roots and columns were not modified, whereas the same responses were abolished when the brainstem was exposed selectively to kynurenic acid, thus suggesting that the responses were carried by long fibres ascending directly to the brainstem. The excitatory and inhibitory synaptic responses are relayed by interneurons located in the brainstem. The ascending excitatory inputs to inhibitory interneurons and, most likely, also to excitatory interneurons, use excitatory amino acid transmission. Inhibitory responses were abolished by adding the glycinergic antagonist strychnine (5 microM) to the physiological solution, thus suggesting that inhibitory interneurons use glycine transmission. The synaptic transmission was depressed by (-)-baclofen, a GABAB agonist, probably acting at a presynaptic site. Taken together, the present results suggest that dorsal root and dorsal column stimulations give rise to disynaptic inhibition and excitation of reticulospinal neurons mediated by excitatory and inhibitory amino acid transmission via brainstem interneurons.

Animals↗

PubMed1993

Anatomical and physiological study of brainstem nuclei relaying dorsal column inputs in lampreys.

The course and sites of termination of dorsal column fibres in the lamprey brainstem are described along with their brainstem relays projecting to reticulospinal neurons. Dorsal column fibres ascend to the brainstem level where they intermingle with cells located in the alar plate close to the obex, a location that is analogous to that of the dorsal column nucleus in other vertebrates. Some dorsal column fibres continue further rostrally where they reach the octavolateralis and octavomotorii nuclei. Finally, a small contingent of fibres reach the cerebellum. Injections of cobalt-lysine into the posterior rhombencephalic reticular nucleus retrogradely label neurons within the dorsal column nucleus and within the octavolateralis and octavomotorii nuclei. Microstimulation of the dorsal column nucleus on either side elicits monosynaptic inhibitory responses in reticulospinal neurons while stimulation of octavolateralis and octavomotorius nuclei elicits excitation. By using intracellular recordings, it was shown that neurons within these alar plate nuclei receive monosynaptic inputs from the dorsal columns. It is thus proposed that disynaptic inputs from dorsal columns to reticulospinal neurons are relayed by these alar plate neurons: inhibition is relayed mainly by neurons in dorsal column nuclei and excitation by neurons in the octavolateralis and octavomotorii nuclei.

Afferent Pathways↗

PubMed1993

Prolidase deficiency: a multisystemic hereditary disorder.

Prolidase deficiency is a rare hereditary disorder with a wide spectrum of clinical manifestations including skin ulcers, eczematous eruptions, characteristic facies, mental retardation, splenomegaly, and susceptibility to infections. We report two new cases of prolidase deficiency. Our patients had the typical manifestations of prolidase deficiency. One also had lupus erythematosus. Prolidase activity was either normal or half-normal in all family members. The skin disease in our patients did not respond to topical glycine/proline ointment or to oral vitamin C.

Adolescent↗

PubMed1993

An immunocytochemical and autoradiographic investigation of the serotoninergic innervation of trigeminal mesencephalic and motor nuclei in the rabbit.

The results of a previous experiment suggest that the cell bodies of many jaw closing muscle spindle afferents in the trigeminal mesencephalic nucleus of the rabbit are phasically inhibited during fictive mastication. The aim of this study was to investigate one possible neurotransmitter system that could be involved in this modulation, serotonin, by use of receptor autoradiography techniques and immunofluorescence combined with retrograde labelling of masseteric spindle afferents and motoneurons. A second objective was to compare the serotonin innervation of neurons in the trigeminal mesencephalic nucleus with that of masseteric motoneurons. Serotoninergic fibres were seen surrounding labelled masseteric spindle afferents, as well as unlabelled neurons, in the trigeminal mesencephalic nucleus. These fibres were close to the cell bodies and sometimes to the axon hillocks of the neurons. Although it has been reported that many neurons of the trigeminal nucleus are multipolar in some species, none of the labelled spindle afferent in this study had more than one process. Throughout the motor trigeminal nucleus, serotonin fibres were found in close proximity with cell bodies and with the proximal portions of axons and dendrites of labelled and unlabelled motoneurons. Serotonin fibres were also seen adjacent to cell bodies and processes of efferent neurons in cell group k. Autoradiography with several tritiated ligands was used to reveal the presence of receptors for serotonin as well as its uptake sites. Only serotonin2 receptors were found to be abundant in the trigeminal mesencephalic nucleus. The motor nucleus and cell group k contained serotonin2 and serotonin3 receptors, as well as serotonin uptake sites. Serotonin1A receptors appear to be absent from both nuclei. The findings suggest that release of serotonin from fibres in close proximity to trigeminal primary afferent somata could modify the transmission of action potentials from muscle spindle receptors during mastication through an action on serotonin2 receptors. In the motor nucleus and cell group k, serotonin may alter neuronal properties through actions on at least two receptor subtypes (serotonin2 and serotonin3).

Action Potentials↗

PubMed1992

Phasic modulation of transmission from vestibular inputs to reticulospinal neurons during fictive locomotion in lampreys.

The aim of this study was to determine whether the transmission from sensory inputs to reticulospinal neurons is modulated during fictive locomotion in lampreys. Reticulospinal neurons play a key role in the control of locomotion; modulation of sensory transmission to these neurons might be of importance for the adaptation of the control they exert during locomotion. In this series of experiments, intracellular synaptic responses of reticulospinal neurons of the posterior rhombencephalic reticular nucleus elicited by electrical stimulation of vestibular nerves on each side were studied during fictive locomotion induced by 50 microM N-methyl-D-aspartate (NMDA). Interestingly, shortly after NMDA had reached the bath and much before locomotor discharges were apparent in the recorded ventral roots, there was a significant depression of the synaptic transmission from vestibular nerves. The effect was reversed by washing out the NMDA and persisted in the isolated brainstem after spinal transection at the first segmental level. As locomotor discharges appeared in the ventral roots, synaptic responses elicited by vestibular nerve stimulation showed a clear phasic modulation of their amplitude during the locomotor cycle. Responses to stimulation of the ipsilateral vestibular nerve were smaller during the ipsilateral burst discharge than during the contralateral activity, whilst responses to stimulation of the contralateral vestibular nerve were minimal during contralateral activity and maximal during ipsilateral activity. This opposite pattern of modulation observed in the same reticulospinal neuron suggests that the phasic modulation of vestibular transmission is not due to changes in the membrane properties of the reticulospinal cell but is produced at a pre-reticular level.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Phasic modulation of vestibulospinal neuron activity during fictive locomotion in lampreys.

This study was aimed at characterizing the activity of vestibulospinal neurons recorded intracellularly during fictive locomotion in lampreys. The majority (78%) of identified vestibulospinal neurons showed rhythmic fluctuations of their membrane potential correlated with locomotor discharges recorded in pairs of rostral ventral roots. Of the rhythmically modulated vestibulospinal cells, most (72%) were maximally depolarized during ipsilateral ventral root discharges and showed a minimum during contralateral activity. Other cells (20%) showed an opposite pattern, that is their peak of depolarization occurred during contralateral activity. Finally, a third category of cells (8%) showed a more complex pattern of activity. Two waves of depolarization could occur per locomotor cycle, one during each burst discharge. The pattern of fluctuation recorded in vestibulospinal neurons appears to be related to the side of the spinal cord onto which the cells are projecting.

Animals↗

PubMed1992

5-Hydroxytryptamine modulates spike frequency regulation in reticulospinal neurons involved in the control of locomotion in lamprey.

To investigate the effects of 5-hydroxytryptamine (5-HT) on reticulospinal neurons involved in the initiation and control of locomotion in lamprey, 5-HT (10 mM) was locally pressure ejected on the dorsal surface of the brainstem during intracellular recordings from identified reticulospinal neurons in the in vitro brainstem-spinal cord preparation. 5-HT induced a reduction of the late afterhyperpolarization (AHP) following the spike due to a reduction of a Ca(2+)-activated K+ current. In addition, 5-HT caused a resting membrane hyperpolarization in a proportion of these cells. Due to the 5-HT induced reduction of the AHP, reticulospinal cells, including those that became hyperpolarized by an application of 5-HT, discharged at a higher rate after 5-HT as a response to the same excitatory drive.

Animals↗

PubMed1992

Evidence that the masticatory muscles receive a direct innervation from cell group k in the rabbit.

These experiments have shown that a group of neurons lateral to the trigeminal motor nucleus innervates the muscles of mastication. The work began to describe the location of digastric last-order interneurons, using the technique of transneuronal labeling with wheatgerm agglutinin-conjugated horseradish peroxide injected into the left digastric muscle of rabbits under general anaesthesia. Four to eight days later, the animals were killed with an overdose of anaesthetic and perfused. Coronal sections of the frozen brainstem were cut at 20 microns thickness and processed for peroxidase activity. Motoneurons in the ventral and caudal divisions of the trigeminal motor nucleus were labeled in all animals as expected. An additional population of neurons located ventrolaterally to the motor nucleus in cell group k were also found to be labeled if the survival time was five days or more. In an attempt to determine whether cell group k neurons were labeled transynaptically, two series of control experiments were carried out. In the first, crystals of fluorescein- and rhodamine-conjugated dextran amines and horseradish peroxidase were applied directly to central ends of cut digastric nerves. In the second, central ends of cut digastric nerves were enclosed in cuffs containing 40-60% horseradish peroxidase solutions. Again, neurons in both the trigeminal motor nucleus and cell group k were labeled suggesting that neurons within cell group k project to the digastric muscle. Similar experiments using dextran amines and wheatgerm peroxidase were carried out on the masseter muscle. Motoneurons in the dorsomedial and rostral half of the trigeminal motor nucleus, as well as primary afferent cell bodies in the mesencephalic nucleus of the trigeminal nerve, were labeled in all experiments. In addition, a population of neurons in cell group k, dorsal to those associated with the digastric muscle, were found to contain each one of the reaction products. Since it is thought that only the wheatgerm agglutinin-conjugated horseradish peroxidase transferred from one neuron to another, we conclude that cell group k neurons provide an additional innervation to the digastric and masseter muscles.

Animals↗

PubMed1991

A new population of neurons with crossed axons in the lamprey spinal cord.

Neurons with contralateral, rostrally and caudally projecting axons were studied in whole mounts of lamprey spinal cord using retrograde labelling techniques with fluorescent dextran-amines, cobalt-lysine or horseradish peroxidase. A previously unknown large population (180-300 cells per hemisegment) of small (less than 25 microns) cells with contralateral projecting axons is described. Their axons extend over less than 5 segments rostrally or caudally. The number of these cells per segment was relatively constant in the rostral half of the spinal cord, but increased significantly in the caudal half. In comparison, medium-sized cells with contralateral axons corresponding to previously identified premotor interneurons were far less numerous (14-21 per hemisegment) and their axons extended more than 5 segments. Contralaterally projecting edge cells (intraspinal stretch receptor neurons) with principal rostral or caudal axons plus short collaterals in the other direction were distributed throughout the length of the spinal cord, whereas large and giant cells with a varied morphology were found in the caudal half.

Animals↗