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L Brodin

Publications and source records attributed to L Brodin.

At least 55 records · Page 3Linked to original sources
PubMed1992

A neuropeptide Y (NPY)-related peptide is present in the river lamprey CNS.

Pancreatic polypeptide (PP)-like material from brain+spinal cord, and retina extracts of Lampetra fluviatilis was studied by HPLC and RIA. The brain+spinal cord extract showed a complex elution profile with multiple peaks of immunoreactivity. The retina extract showed a much simpler pattern with a single significant peak along with a trace of a second peak corresponding to the latest and penultimate peaks in the brain extract. Twenty-one out of 36 residues could be sequenced from the latest eluting peak in the brain extract. This sequence showed 81% identity with porcine neuropeptide (NPY) suggesting that both the brain/spinal cord and retina of the river lamprey contain a peptide homologous to NPY.

Amino Acid Sequence↗
PubMed1992

A computer-based model for realistic simulations of neural networks. II. The segmental network generating locomotor rhythmicity in the lamprey.

1. To analyze the function of the spinal interneuronal network generating locomotion in the lamprey CNS, a vertebrate model system, we performed computer simulations with realistic model neurons possessing the essential properties of their biological counterparts. 2. The segmental network has been simulated by modeling experimentally established types of neurons with their specific membrane properties and synaptic interconnections. Fictive locomotor activity, which can be experimentally induced by elevating the background excitability by bath application of excitatory amino acids, was simulated by opening membrane conductances for kainate/alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) or N-methyl-D-aspartate (NMDA) receptors. Kainate/AMPA receptor activation induced a rhythm in the middle and upper part of the physiological burst frequency range, whereas NMDA receptor activation evoked bursting in the lower part of the range, which corresponds well to earlier experimental findings. 3. Several factors contributing to the termination of the burst were studied and their interaction was assessed in simulations of the network. 1) The summation of spike afterhyperpolarizations (late AHPs), leading to adaptation of the discharge, acts as a primary burst-terminating factor at lower rates of kainate/AMPA-induced bursting, and it also interacts with the NMDA-induced oscillatory membrane properties during slow rhythmicity. 2) The termination of the depolarized NMDA plateau is another important factor during NMDA-evoked rhythmicity. 3) The synaptic inhibition from lateral interneurons to the interneurons mediating reciprocal inhibition is important at higher rates of kainate/AMPA-induced bursting. 4. The mechanism of action of 5-hydroxytryptamine (5-HT) on the lamprey segmental network was further investigated by simulation. 5-HT is known to lower the burst frequency during fictive locomotion and also to decrease the conductance through the Ca(2+)-dependent K+ channels, and thereby the size of the late AHP that follows the action potential. Decreasing this conductance in the network simulations resulted in a lesser amount of AHP summation and thereby less frequency adaptation during the burst, longer bursts, and a lower locomotor frequency. Thus the selective action of 5-HT on the Ca(2+)-dependent K+ channels, and hence on the AHP, can account for the modulatory effect on the fictive locomotor rhythm seen experimentally. 5. The results demonstrate that the present simulation of the segmental network can account for essential features of the motor pattern seen experimentally during lamprey locomotion.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1992

Immunogold quantification of glutamate in two types of excitatory synapse with different firing patterns.

A quantitative immunocytochemical method was used to study the regional levels of glutamate in two types of lamprey (Ichtyomyzon unicuspis) axon, which both activate excitatory amino acid receptors, but which when active exhibit different firing patterns. Giant reticulospinal axons fire in brief bursts, while dorsal column axons, mainly belonging to cutaneous afferents, show a sustained firing at high frequency. In both types of axon, clusters of synaptic vesicles showed a strong accumulation of glutamate immunogold labeling, and the density of gold particles correlated strictly with the packing density of synaptic vesicles. The most densely packed vesicle areas had a particle density corresponding to a concentration of fixed glutamate of about 30 mM in coprocessed glutamate conjugates, suggesting an intravesicular glutamate concentration of at least 60 mM. The level of labeling in axoplasmic matrix was considerably lower, but differed significantly between the two types of axon. Dorsal column axons showed a particle density in axoplasmic matrix that was approximately four times higher than that in giant reticulospinal axons. The mitochondrial glutamate labeling was also significantly stronger in the dorsal column axons. In addition, the number of mitochondrial profiles surrounding vesicle clusters was about four times higher in dorsal column synapses than in reticulospinal synapses. Antisera to aspartate, GABA, glutamine, and homocysteate failed to produce a specific labeling of synaptic vesicle clusters in reticulospinal or dorsal column axons. In conjunction with previous demonstrations of a stimulus-induced vesicle depletion in giant reticulospinal synapses (Wickelgren et al., 1985), these results imply that glutamate is released from synaptic vesicles. The different extravesicular glutamate levels in reticulospinal axons and dorsal column axons may relate to different requirements for the refilling of synaptic vesicles in these functionally divergent neurons.

Animals↗
PubMed1992

Differential expression of mRNAs for neuropeptide Y-related peptides in rat nervous tissues: possible evolutionary conservation.

Neuropeptide Y (NPY) is the only member of its peptide family that has been isolated from the mammalian CNS. We have recently found that two different NPY-related molecules are present in the CNS of a cyclostome, the river lamprey (Lampetra fluviatilis) (Söderberg et al., 1991). Here we show that this is also true for the rat CNS, by demonstrating expression of peptide YY (PYY) mRNA in brainstem neurons distinct from those neurons that express NPY mRNA. Dissimilar oligonucleotide DNA probes complementary to 3' untranslated regions of the rat PYY, NPY, and pancreatic polypeptide (PP) mRNA were used in in situ hybridization experiments on sections of rat brain and spinal cord, visceral organs, and peripheral nerve ganglia. The PYY probe hybridized with two populations of neurons in the brainstem: one dispersed along the midline in the rostral medulla and another in the lateral caudal medulla (A1 region). No additional labeling was detected in the remainder of the neuraxis. In the periphery, PYY hybridization was seen only in endocrine cells of the colon, and not in sympathetic ganglia or the adrenal gland, suggesting that previous observations of PYY immunoreactivity in these latter structures were due to antibody cross-reactivity with NPY. The NPY probe did not hybridize with cells on the midline region that contains PYY neurons, but it did label large numbers of neurons throughout the neuraxis. No expression of PP mRNA was detected in the CNS. Northern blot analysis failed to detect PYY mRNA in the CNS, further supporting the observation that PYY is only expressed by a discrete collection of CNS neurons. The anatomy of PYY- and NPY-expressing cells in the CNS and gut shows a striking similarity between rat and lamprey (Brodin et al., 1989), vertebrates that diverged evolutionarily about 450 million years ago, suggesting that both peptide systems have been conserved throughout vertebrate evolution.

Animals↗
PubMed1991

A computer based model for realistic simulations of neural networks. I. The single neuron and synaptic interaction.

The use of computer simulations as a neurophysiological tool creates new possibilities to understand complex systems and to test whether a given model can explain experimental findings. Simulations, however, require a detailed specification of the model, including the nerve cell action potential and synaptic transmission. We describe a neuron model of intermediate complexity, with a small number of compartments representing the soma and the dendritic tree, and equipped with Na+, K+, Ca2+, and Ca2+ dependent K+ channels. Conductance changes in the different compartments are used to model conventional excitatory and inhibitory synaptic interactions. Voltage dependent NMDA-receptor channels are also included, and influence both the electrical conductance and the inflow of Ca2+ ions. This neuron model has been designed for the analysis of neural networks and specifically for the simulation of the network generating locomotion in a simple vertebrate, the lamprey. By assigning experimentally established properties to the simulated cells and their synapses, it has been possible to verify the sufficiency of these properties to account for a number of experimental findings of the network in operation. The model is, however, sufficiently general to be useful for realistic simulation also of other neural systems.

Computer Simulation↗
PubMed1991

Computer simulations of N-methyl-D-aspartate receptor-induced membrane properties in a neuron model.

1. To evaluate the role of N-methyl-D-aspartate (NMDA) receptors in simulations of the lamprey spinal locomotor network, we developed a computer-simulated electrical model of a neuron that contains NMDA channels in addition to voltage-gated Na+, K+, and Ca2+ channels and Ca(2+)-activated K+ channels [K(Ca) channels]. 2. The voltage dependence of the Mg2+ block of the Na(+)-K+ current flow through the NMDA channel was modeled according to a scheme of open-channel block. To account for the regulation of K(Ca) channels by NMDA and membrane voltage, we modeled two separate Ca2+ pools that had different voltage dependencies and dynamics. 3. Pacemaker-like membrane potential oscillations could be elicited in the model neuron, which resembled those observed experimentally in the presence of bath-applied NMDA and tetrodotoxin. The effect of changing different channel parameters were tested to determine under which conditions such membrane potential oscillations could occur. 4. The oscillation amplitude was determined by the potential levels at which the NMDA channels and voltage-dependent K+ channels, respectively, were activated. The oscillation frequency and the relative durations of the de- and hyperpolarized phases of the oscillations were determined by the balance between the depolarizing (NMDA channels) and hyperpolarizing [K(Ca) channels] currents. 5. Simulated alterations of the Mg2+ concentration and the K+ conductance as well as injection of constant current caused changes of the oscillations corresponding to those observed experimentally. The de- and hyperpolarizing phases could be reset by brief current pulses. 6. We conclude that the present model can account for the effects of bath-applied NMDA on spinal neurons. This permits an incorporation of NMDA-receptor-mediated properties in simulation models of the lamprey locomotor network.

Action Potentials↗
PubMed1990

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↗
PubMed1990

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↗
PubMed1989

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.

Animals↗
PubMed1989

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↗
PubMed1989

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↗
PubMed1989

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↗
PubMed1989

Three-dimensional imaging of neurons by confocal fluorescence microscopy.

The study of neuronal architecture by means of confocal laser microscopy is described. Optical serial sectioning has been performed on whole-mount specimens, and the resulting stacks of digitally recorded images have been processed with the help of a computer. Specimen preparation is described, as well as the instrument and its performance. It is shown that the limits in photometric quality are set by photon quantum noise. As both light absorption and scattering was low in the studied specimens, the maximum scanning depth was limited mainly by the working distance of the objectives. Compared with traditional methods, confocal microscopy in combination with digital image processing has the following advantages: (1) a truly three-dimensional (3-D) reconstruction is obtained, (2) the specimen remains intact, (3) a higher resolution can be obtained, (4) the process is automated and less time-consuming and (5) various kinds of data processing are possible.

Animals↗
PubMed1989

Multiple forms of pancreatic polypeptide-related compounds in the lamprey CNS: partial characterization and immunohistochemical localization in the brain stem and spinal cord.

Although neuropeptide Y (NPY) is established as a transmitter in many regions of the nervous system, the role of other peptides of the pancreatic polypeptide (PP) family in the CNS is obscure. This study provides evidence that PP-like peptides in the "primitive" CNS of a cyclostome are composed of different molecular forms, which are stored in separate neuronal populations with apparently different functions. PP-like material was detected in extracts of brain and spinal cord from Lampetra fluviatilis by radioimmunoassay (RIA) using an antiserum to the C-terminal hexapeptide of mammalian PP. The PP-immunoreactive material consisted of several molecular forms, as shown by its complex elution profile on high-performance liquid chromatography (HPLC). The cellular distribution of PP-like immunoreactivity was studied with indirect immunofluorescence histochemistry using antisera toward porcine peptide YY (PYY), porcine neuropeptide Y (NPY), and bovine (BPP), rat (RPP), and avian (APP) pancreatic polypeptide. Adjacent sections from the brain stem and spinal cord of L. fluviatilis and Ichthyomyzon unicuspis, incubated with the different antisera, displayed 2 main patterns of PP immunoreactivity. The PYY and RPP antisera labeled groups of neurons and fibers in the rhombencephalic and mesencephalic reticular formation. One of the PYY/RPP-ir cell groups, located in the anterior rhombencephalic reticular nucleus, had a projection to the dorsolateral spinal cord. Fibers of this reticulospinal system were in close apposition to dendrites of intracellularly stained spinal motoneurons and sensory relay interneurons, indicating that they may receive PPergic input. In contrast, antisera to NPY and APP labeled local neurons systems in the spinal dorsal horn, in the lateral parts of the brain stem, including the rhombencephalic alar plate, and in the retina. The BPP antiserum recognized the NPY/APP as well as the PYY/RPP immunoreactive neuron systems, further supporting that they both contain PP-like peptides.

Animals↗
PubMed1988

The effect of an uptake inhibitor (dihydrokainate) on endogenous excitatory amino acids in the lamprey spinal cord as revealed by microdialysis.

Microdialysis was utilized to test the effects of the uptake inhibitor dihydrokainate (DHK) on endogenous amino acid levels in the lamprey spinal cord in vitro. The level of L-glutamate increased markedly (165%) in the presence of DHK, whereas the level of the glutamate precursor L-glutamine decreased (53%). Since DHK can potentiate or evoke fictive locomotion in the lamprey spinal cord, it is suggested that L-glutamate is released by neurons which take part in the activation of the spinal locomotor network.

Animals↗
PubMed1988

Immunohistochemical studies of cholecystokininlike peptides and their relation to 5-HT, CGRP, and bombesin immunoreactivities in the brainstem and spinal cord of lampreys.

The distribution of cholecystokinin (CCK)-like immunoreactivity in the brainstem and spinal cord of lampreys was studied by using CCK antisera with different properties. In the spinal cord, three separate systems reacted with CCK antisera: (1) A ventral and lateral fiber system descending from a group of neurons in the posterior reticular nucleus of the rhombencephalon was labeled by both a C-terminal-directed CCK antiserum and a monoclonal CCK antibody. (2) A dorsal root-dorsal column system of fibers originating from cell bodies in the dorsal root ganglia was labeled only by the C-terminal CCK antiserum. This CCK immunoreactivity could be abolished by preabsorption with calcitonin-gene-related peptide (CGRP), suggesting that it was due to cross-reactivity with a CGRP-like peptide. This system also contained 5-hydroxytryptamine (5-HT)-, bombesin-, and CGRP-like immunoreactivities. (3) An intraspinal system of 5-HT neurons was labeled with an antiserum to the midportion of CCK-33 but not by the other CCK antisera. The CCK labeling of this system was difficult to reduce by preabsorption with CCK peptide and thus appeared to be nonspecific. Groups of cell bodies in the middle reticular nucleus of the rhombencephalon, the reticular nucleus of the mesencephalon, and the hypothalamus were labeled by both the C-terminal and the monoclonal CCK antisera. The gut contained two types of CCK-like immunoreactivity, one of which appeared to be due to cross-reactivity with CGRP. A biochemical analysis showed that the content of CCK was low in the spinal cord compared to the brain, and these results agreed with the immunohistochemical findings.

Animals↗
PubMed1988

Possible target neurons of the reticulospinal cholecystokinin (CCK) projection to the lamprey spinal cord: immunohistochemistry combined with intracellular staining with lucifer yellow.

A subpopulation of reticulospinal neurons in the posterior rhombencephalic reticular nucleus in lamprey contains a cholecystokinin (CCK)-like peptide. Varicosities of these neurons, visualized by immunohistofluorescence, were found to be in close apposition to Lucifer yellow-filled spinal motoneurons and giant relay interneurons. Certain other types of interneurons and primary sensory neurons had no, or very few, close contacts with CCK-immunoreactive boutons.

Animals↗