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S Grillner

Publications and source records attributed to S Grillner.

At least 145 records · Page 8Linked to original sources

Organization of the six motor nuclei innervating the ocular muscles in lamprey.

The topography of motoneurons supplying each of the six ocular muscles of the lamprey, Lampetra fluviatilis, was studied by selective application of HRP to the cut nerves of identified muscles. In addition, the distributions of motoneuron populations to both eyes were studied simultaneously with fluorescein and rhodamine coupled dextran-amines (FDA and RDA) applied to cut ocular muscle nerves of either side. The motoneuron pool of the caudal oblique muscle is represented bilaterally in the trochlear (N IV) motor nucleus. The dorsal rectus muscle is innervated from a contralateral group of oculomotor (N III) motoneurons and the remaining four muscles exclusively from the ipsilateral side (N III and N VI). The inferior and posterior rectus muscles are both innervated by the abducens nerve. In contrast to all jawed vertebrates, only three eye muscles (the dorsal rectus, rostral rectus, and rostral oblique) are innervated by the oculomotor nerve in lampreys (N III). Lampreys have a motor nucleus similar to the accessory abducens nucleus previously described only in tetrapods. They lack the muscle homologous to the nasal rectus muscle of elasmobranchs and the medial rectus muscle of osteognathostomes. The distribution of the dendrites of different groups of motoneurons was studied and is considered in relation to inputs from tectum and the different cranial nerves.

Animals↗

5-hydroxytryptamine immunoreactive varicosities in the lamprey spinal cord have no synaptic specializations--an ultrastructural study.

The distribution and fine structure of 5-hydroxytryptamine (5-HT) immunoreactive cell bodies and axonal varicosities have been studied in the lamprey spinal cord, using the peroxidase-antiperoxidase (PAP) immunohistochemical technique and subsequent analysis of ultrathin serial sections. Immunostained cell bodies were found in the ventral spinal cord close to the central canal. Immunostained varicosities were found throughout the spinal cord with the highest density in the ventromedial plexus and the dorsal horn. Only large granular vesicles could be clearly distinguished in immunostained cell bodies and varicosities, but it was concluded based on a comparison with unstained normal tissue that these boutons also contained small, pleomorphic agranular vesicles. Immunoreactive varicosities were studied in the ventromedial plexus, the dorsal horn, the dorsal column, the dorsolateral and ventrolateral funiculi and the grey matter. No morphological differences could be observed between varicosities in the different loci. The varicosities were in no case seen to make synaptic contact with surrounding neuronal elements, even when followed through serial sections. Consequently, 5-HT released from boutons in all parts of the spinal cord could be expected to act on 5-HT receptors located on nearby as well as distant receptors.

Animals↗

Distribution of histaminergic neurons in the brain of the lamprey Lampetra fluviatilis as revealed by histamine-immunohistochemistry.

An antiserum against conjugated histamine was used to study the distribution of histaminergic neurons in the CNS of the lamprey Lampetra fluviatilis. Numerous histamine-immunoreactive cell bodies were detected in the dorsal and ventral hypothalamic nuclei and in the adjacent postinfundibular commissural nucleus. Histamine-immunoreactive fibers of high density were present in the ventral hypothalamus, and fibers could also be traced dorsally from the hypothalamus to the corpus striatum and septal nucleus where they appeared to terminate in dense plexuses. Another, smaller group of histamine-immunoreactive perikarya was observed in the border area between mesencephalon and rhombencephalon, near the caudal pole of the mesencephalic reticular nucleus. Sparsely distributed histamine-immunoreactive fibers were present in the ventral mesencephalon. The distribution of histaminergic neurons in cyclostomes, which diverged very early from the main vertebrate line, shows similarities with the corresponding systems in the CNS of amphibians and mammals, which suggests that histaminergic neuronal systems are phylogenetically old and have been conserved during evolution.

Animals↗

Three types of GABA-immunoreactive cells in the lamprey spinal cord.

Polyclonal antisera raised against conjugated GABA were used to study the distribution of GABAergic neurons in the spinal cords of lampreys (Lampetra fluviatilis and Ichtyomyzon unicuspis) using immunofluorescence and peroxidase-antiperoxidase techniques. Three morphologically distinct types of GABA-immunoreactive (GABA-ir) cell bodies were observed, multipolar neurons in the lateral grey cell column, apparently bipolar cells in the ventral aspect of the dorsal horn, and small liquor-contacting cells surrounding the central canal. A high density of immunoreactive fibers of spinal origin were present in the lateral and ventral funiculi, whereas the dorsal column had a relatively low density. Dense GABA-ir plexuses were situated in the lateral spinal margin, and in the dorsal part of the dorsal horn. A chronic lesion of the rostral spinal cord did not result in any observable loss of GABA-ir fibers below or above the lesion, suggesting that the 3 types of segmental GABA-ir neurons are the main sources of the GABAergic innervation of the lamprey spinal cord.

Animals↗

CNQX and DNQX block non-NMDA synaptic transmission but not NMDA-evoked locomotion in lamprey spinal cord.

The motor pattern underlying locomotion in the lamprey is activated and maintained by excitatory amino acid neurotransmission. The quinoxalinediones 6,7-dinitroquinoxaline-2,3-dione (DNQX) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) are potent and selective antagonists of non-N-methyl-D-aspartate (NMDA) receptors in the mammalian central nervous system. In the lamprey, these compounds are now shown to block fast excitatory synaptic potentials elicited in neurones of the spinal ventral horn. They selectively antagonise responses to the application of selective kainate and quisqualate receptor agonists (kainate and alpha-amino-3-hydroxy-5-methyl-4-isoxalone (AMPA)) but do not influence NMDA receptor-mediated responses. Additionally, it is shown that the activation of NMDA receptors is sufficient to elicit and maintain fictive locomotion after blockade of non-NMDA receptors with either DNQX or CNQX. Conversely, activation of quisqualate receptors with AMPA, but not quisqualate leads to fictive locomotion with properties much like that activated by kainate.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

Excitatory amino acids and synaptic transmission: the evidence for a physiological function.

For 30 years physiological techniques have been used to investigate excitatory amino acids as neurotransmitters. In the last ten years progress on the definition of receptor subtypes and the availability of more selective agonists and antagonists has fuelled physiological, neurochemical and histochemical approaches to elucidating the involvement of excitatory amino acids at synaptic sites throughout the vertebrate CNS. Here Max Headley and Sten Grillner assess the advances made in defining the roles of excitatory amino acids as functional transmitters, taking examples mainly from studies on the spinal cord, and comment on the limitations of the types of approach that are used in such studies.

Amino Acids↗

Intersegmental co-ordination of undulatory movements--a "trailing oscillator" hypothesis.

Intersegmental co-ordination in undulatory locomotion was investigated in the lamprey spinal cord in vitro. It is characterized by a constant phase lag between the activation of consecutive segments. By increasing the excitability (NMDA level) locally, a "travelling wave" could be induced to start in any segment and to be conducted in both rostral and caudal directions. This leading segment, the one with the highest excitability, determined the frequency of all other segments, and the ascending and descending couplings were functionally symmetric. The intersegmental phase lag can be explained by a simple neural mechanism in which a leading oscillator will entrain the adjacent "trailing" segments, which in turn entrain their neighbours and so forth.

Animals↗

Activation of N-methyl-D-aspartate (NMDA) receptors augments repolarizing responses in lamprey spinal neurons.

Current- and voltage-clamp techniques were used to analyze the mechanisms underlying the repolarization during N-methyl-D-aspartate (NMDA)-induced, tetrodotoxin-resistant pacemaker-like oscillations in lamprey spinal neurons. Long-lasting depolarizing current pulses (15-40 mV, 50-400 ms, tetrodotoxin and tetraethylammonium present) were followed by hyperpolarizing afterpotentials even when NMDA receptors were blocked, but they were markedly enhanced by application of N-methyl-D,L-aspartate (NM(DL)A). The afterpotentials were depressed by replacing Ca2+ with Ba2+. During voltage-clamp NM(DL)A enhanced a Ba2+-sensitive outward tail current following voltage steps of 15-40 mV. The outward current remained after injection of Cl-, as did the NMDA-induced membrane potential oscillations observed under current-clamp. These results suggest that the repolarization during NMDA-induced oscillations is due to Ca2+ entry both via NMDA-gated channels and conventional voltage-gated Ca2+ channels, leading to an activation of Ca2+-dependent K+ channels. The afterhyperpolarization following single action potentials, which is also due to Ca2+-dependent K+ channels, was not significantly altered by NMDA receptor activation, suggesting a different location of the Ca2+ entry during the two conditions in relation to the location of the activated Ca2+-dependent K+ channels.

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Visuomotor coordination in reaching and locomotion.

Locomotion and reaching have traditionally been regarded as separate motor activities. In fact, they may be closely connected both from an evolutionary and a neurophysiological viewpoint. Reaching seems to have evolved from the neural systems responsible for the active and precise positioning of the limb during locomotion; moreover, it seems to be organized in the spinal cord. The motor cortex and its corticospinal outflow are preferentially engaged when precise positioning of the limb is needed during locomotion and are also involved during reaching and active positioning of the hand near objects of interest. All of these motor activities require visuomotor coordination, and it is this coordination that could be achieved by the motor cortex and interconnected parietal and cerebellar areas.

Animals↗

Increase in endogenous 5-hydroxytryptamine levels modulates the central network underlying locomotion in the lamprey spinal cord.

To investigate the effects of an endogenous release of serotonin (5-HT) in the lamprey spinal cord, in vitro, the 5-HT uptake-blocker citalopram (1-10 microM) was added to the bathing solution. Samples taken from the physiological solution through high-performance liquid chromatography (HPLC) showed that 5-HT was released from the spinal cord. To study the effect of this endogenous release of 5-HT on the spinal network generating locomotion, 'fictive locomotion' was induced by bath application of N-methyl-D-aspartate (NMDA, 100 microM). It elicited a steady locomotor rhythm between 0.2 and 2.5 Hz. The effects of citalopram were the following: (1) the locomotor frequency slowed down, (2) the intensity of the ventral root bursts was increased and (3) the intersegmental phase lag was prolonged. The effects of citalopram and thus presumably of an endogenous release of 5-HT were similar to what has previously been observed during bath application of 5-HT. In the latter case the conditions are different, since all 5-HT receptors regardless of their location in relation to the 5-HT-containing boutons will be affected in a similar way.

Action Potentials↗

Phasic modulation of reticulospinal neurones during fictive locomotion and other types of spinal motor activity in lamprey.

The intracellular activity of different types of reticulospinal neurones was studied during fictive locomotion and other types of spinal motor activity in an in vitro preparation of the lamprey brainstem-spinal cord. The examined neurones included large Müller cells of the rhombencephalic and mesencephalic reticular formation, the Mauthner cell, and neurones in the posterior rhombencephalic reticular nucleus with different sizes and conduction velocities. During bouts of fictive swimming initiated spontaneously or by stimulation of the trigeminal nerve or spinal cord, the Müller cells were depolarized and fired action potentials. Bulbar Müller cells in addition showed a phasic modulation of membrane potential with excitation in phase with ipsilateral motoneurones of the rostral spinal cord. The Mauthner cell was depolarized in phase with contralateral motoneurones. Many neurones in the posterior rhombencephalic reticular nucleus showed modulation in phase with ipsilateral motoneurones during fictive swimming. Such oscillations were observed in both fast-conducting neurones, located mainly in the medial part of the nucleus, and slower conducting cells with a more lateral distribution. All examined reticulospinal neurones showed a strong coupling also with other types of spinal motor activity, such as slow alternating bursting and synchronous bilateral ventral root bursts, but the reticulospinal activity had no correlation with respiratory activity recorded from the Xth nerve. The consequences of a phasic reticulospinal activity during locomotion are discussed.

Action Potentials↗

The role of spinal cord inputs in modulating the activity of reticulospinal neurons during fictive locomotion in the lamprey.

Lamprey reticulospinal neurons are rhythmically modulated during fictive swimming. The present study examines the possibility that this modulation may originate from the spinal cord locomotor networks rather than from the brainstem. To test this, the in vitro preparation of the lamprey brainstem-spinal cord was separated into two compartments which could be exposed to different chemical environments. Locomotor activity was induced pharmacologically in the caudal spinal cord compartment and reticulospinal (RS) neurons from the posterior rhombencephalic reticular nucleus (PRRN) were recorded intracellularly in the rostral compartment containing normal lamprey Ringer. Under these conditions, the membrane potential of RS neurons showed clear rhythmic oscillations which are correlated with the ongoing locomotor activity in the caudal spinal cord bath, although no locomotor discharges were present in the ventral roots of the rostral bath. Such oscillations were not present in the absence of locomotion. These results indicate that the spinal cord locomotor networks can contribute to the rhythmic oscillations which occur in RS neurons during fictive locomotion. Moreover, the latter oscillations of membrane potential are due to both phasic excitation and Cl- -dependent inhibition in the opposite phase.

Action Potentials↗

Further evidence for excitatory amino acid transmission in lamprey reticulospinal neurons: selective retrograde labeling with (3H)D-aspartate.

The distribution of radiolabeled neurons in the brain stem of Lampetra fluviatilis was studied following unilateral injections of (3H)D-aspartate in the rostral spinal cord. After survival periods of 1-3 days, labeled perikarya were present within and nearby the posterior, middle, and anterior rhombencephalic reticular nuclei and in the mesencephalic reticular nucleus. The highest number of (3H)D-aspartate labeled cell bodies were present in the posterior rhombencephalic reticular nucleus. The labeled reticulospinal neurons were distributed mainly ipsilateral to the injection site and included the giant Müller cells as well as medium-sized and small neurons. Contralateral labeling occurred in cell bodies scattered along the lateral margin of the rhombencephalic reticular formation, the most rostral of these contralaterally projecting neurons being the Mauthner cell. The (3H)D-aspartate labeling correlates with previous electrophysiological studies showing that lamprey reticulospinal neurons utilize excitatory amino acid transmission.

Animals↗

Efferents to the labyrinth of the river lamprey (Lampetra fluviatilis) as revealed with retrograde tracing techniques.

The cells of origin of centrifugal fibres to the labyrinth were examined in a jawless vertebrate, the lamprey, with several retrograde tracers (horseradish peroxidase (HRP), fluorescein isothiocyanate (FITC) and rhodamine coupled dextran-amines (FDA and RDA), and cobalt-lysine) that had been applied to the eighth nerve. With all 3 techniques, up to 20 cells were consistently labelled ipsilaterally, adjacent to the Mauthner cell; none were labelled on the contralateral side. The axons of the efferent cells emanate either from lateral dendrites or the perikaryon and course laterally towards the ipsilateral eighth nerve root. The dendrites ramify widely in the basal plate and may thus receive diverse inputs. The same tracers were applied to the lateral-line nerve and afferent fibres were labelled but no efferent cells, thus confirming previous reports. These data suggest that lampreys have an efferent innervation to the labyrinth but not to the lateral line.

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Mechanisms underlying the serotonergic modulation of the spinal circuitry for locomotion in lamprey.

The central nervous system of the lamprey contains serotonergic (5-hydroxytryptamine, 5-HT) neurones both in the spinal cord and in the brainstem. Endogenously released 5-HT from these systems modulates the pattern of fictive locomotion induced in the in vitro preparation; the burst rate is lowered and burst discharges become longer and of higher intensity. Local application of 5-HT, mimicking activation of the 5-HT systems, has a specific effect on the late phase of the afterhyperpolarization (AHP) in motoneurones and interneurones. 5-HT markedly reduces the amplitude of the late AHP without affecting passive membrane properties or the shape or threshold of the action potential. This 5-HT effect appears to result from a direct action on the calcium-dependent potassium channels underlying the late phase of the AHP. A reduction of the amplitude of the AHP will result in altered spike discharge characteristics, with potentiation of the response (discharge rate) to a given excitatory input in all neurones influenced by 5-HT. It is suggested that the modulatory effect of 5-HT on fictive locomotion can be attributed to its action on the late AHP and thereby to the potentiation of excitability in excitatory and inhibitory interneurones in the generator circuitry. This has been further corroborated in computer simulation studies of a network model, where the action of 5-HT was simulated by decreasing AHP amplitude, resulting in a slowing of the rhythm analogous to the effect demonstrated experimentally.

Animals↗

Effects of 5-hydroxytryptamine on the afterhyperpolarization, spike frequency regulation, and oscillatory membrane properties in lamprey spinal cord neurons.

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

Action Potentials↗

Identification of excitatory interneurons contributing to generation of locomotion in lamprey: structure, pharmacology, and function.

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

Action Potentials↗