Russell H. Hill

Neural networks that co-ordinate locomotion and body orientation in lamprey

The networks of the brainstem and spinal cord that co-ordinate locomotion and body orientation in lamprey are described. The cycle-to-cycle pattern generation of these networks is produced by interacting glutamatergic and glycinergic neurones, with NMDA receptor-channels playing an important role at lower rates of locomotion. The fine tuning of the networks produced by 5-HT, dopamine and GABA systems involves a modulation of Ca2+-dependent K+ channels, high- and low-threshold voltage-activated Ca2+ channels and presynaptic inhibitory mechanisms. Mathematical modelling has been used to explore the capacity of these biological networks. The vestibular control of the body orientation during swimming is exerted via reticulospinal neurones located in different reticular nuclei. These neurones become activated maximally at different angles of tilt.

The intrinsic function of a motor system — from ion channels to networks and behavior

The forebrain, brainstem and spinal cord contribution to the control of locomotion is reviewed in this article. The lamprey is used as an experimental model since it allows a detailed cellular analysis of the neuronal network underlying locomotion. The focus is on cellular mechanisms that are important for the pattern generation, as well as different types of pre- and postsynaptic modulation. This experimental model is bridging the gap between the molecular and cellular level to the network and behavioral level.

Transmitters, membrane properties and network circuitry in the control of locomotion in lamprey

Abstract The lamprey brainstem and spinal cord can be maintained in vitro . It is a simple vertebrate preparation with comparatively few neurones. The neural correlates of different patterns of behaviour can be elicited in this in-vitro preparation. The subject of this review is the neuronal organization underlying locomotion, and, in particular, the role of different types of interneurones and their transmitters and mode of synaptic interaction. Excitatory amino acids, glycine, GABA, 5-HT, tachykinins and CCK have been implied as putative transmitters. The activation of one type of excitatory amino acid receptor, the NMDA receptor, can elicit TTX-resistant pacemaker-like membrane, potential oscillations. 5-HT can exert indirectly a potentiating effect via a depression of the postspike after-hyperpolarization (Ca 2+ -dependent potassium channels).

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

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

Interferon-γ induces characteristics of central sensitization in spinal dorsal horn neurons in vitro

&NA; Hyperexcitability of spinal dorsal horn neurons, also known as ‘central sensitization’, is a component of pain associated with pathological conditions in the nervous system. The aim of the present study was to analyze if the pro‐inflammatory cytokine, interferon‐&ggr; (IFN‐&ggr;), which can be released for extended periods of time in the nervous system during inflammatory and infectious events, can alter synaptic activity in dorsal horn neurons and thereby contribute to such hyperexcitability. Treatment of cultured dorsal horn neurons with IFN‐&ggr; for 2 weeks resulted in a significantly reduced clustering of &agr;‐amino‐3‐hydroxy‐5‐methylisoxazole (AMPA) receptor subunit 1 (GluR1) that was dependent on nitric oxide. The neurons displayed an increased frequency and amplitude of excitatory postsynaptic currents (EPSCs) upon IFN‐&ggr; treatment. Treated dorsal horn neurons also exhibited increased responsiveness to stimulation of dorsal root ganglia (DRG) axons in a two‐compartment model. Furthermore, disinhibition by the GABAA receptor antagonist picrotoxin (PTX) significantly increased EPSC frequency and induced bursting in untreated cultures but did not significantly increase the frequency in treated neurons, which displayed bursting even without PTX. GABAA agonists reduced activity more strongly in treated cultures and immunochemical staining for GABAA receptors showed no difference from controls. Since GluR1‐containing AMPA receptors (AMPARs) occur predominantly on inhibitory neurons in the dorsal horn, we suggest that the IFN‐&ggr;‐mediated increase in spontaneous activity and responsiveness to DRG axon stimulation, decrease in sensitivity to PTX and tendency for EPSC bursting result from a reduced expression of GluR1 on these neurons and not from a reduction in active GABAA receptors in the network. IFN‐&ggr; thereby likely causes disinhibition of synaptic activity and primary afferent input in the dorsal horn, which consequently results in central sensitization.

The action of 5-HT on calcium-dependent potassium channels and on the spinal locomotor network in lamprey is mediated by 5-HT1A-like receptors.

5-HT has a powerful modulatory action on the firing properties of single neurons as well as on locomotor activity. In lamprey, 5-HT increases the neuronal firing frequency in spinal neurons by reducing the conductance in Ca(2+)-dependent K+ channels (KCa) underlying the slow afterhyperpolarization (sAHP), and it also lowers the burst frequency of the spinal locomotor network. To elucidate which type of 5-HT receptor mediates these effects, different specific receptor agonists and antagonists were applied during intracellular current clamp recordings and during NMDA-induced fictive locomotion in the lamprey spinal cord in vitro preparation. The 5-HT1A receptor agonist 8-OH-DPAT ((+/-)-8-hydroxy-dipropylaminotetralin hydrobromide), the 5-HT1 receptor agonist 5-CT (5-carboxyamidotryptamine maleate) and the 5-HT2 receptor agonist alpha-CH3-5-HT (alpha-methylserotonin maleate) all reproduced the actions of 5-HT at both the cellular and the network levels. The effects of all agonists were completely or partially blocked by the 5-HT1A and 5-HT2 receptor antagonist spiperone (spiroperidol hydrochloride) while selective 5-HT2 receptor antagonists were ineffective. The selective 5-HT1A receptor antagonist S(-)-UH301 (S(-)-5-fluoro-8-hydroxy-dipropylaminotetralin hydrochloride) also counteracted the effect of 5-HT on the sAHP. 5-HT3 and 5-HT4 receptor agonists and antagonists were without effects. The intracellular coupling mechanism was not sensitive to pertussis toxin nor to the cAMP dependent protein kinase blocker (Rp)-cAMPS.(ABSTRACT TRUNCATED AT 250 WORDS)

Age-related changes in electrophysiological properties of the mouse suprachiasmatic nucleus in vitro

Endogenous biological rhythms are altered at several functional levels during aging. The major pacemaker driving biological rhythms in mammals is the suprachiasmatic nucleus of the hypothalamus. In the present study we used tissue slices from young and old mice to analyze the electrophysiological properties of the retinorecipient ventrolateral part of the suprachiasmatic nucleus. Loose patch and whole-cell recordings were performed during day and night. Both young and old mice displayed a significant variation between day and night in the mean firing rate of suprachiasmatic nucleus neurons. The proportion of cells not firing spontaneous action potentials showed a clear day/night rhythm in young but not in old animals, that had an elevated number of such silent cells during the day compared to young animals. Analysis of firing patterns revealed a more regular spontaneous firing during the day than during the night in the old mice, while there was no difference between day and night in young animals. The frequency of spontaneous inhibitory postsynaptic currents was reduced in ventrolateral suprachiasmatic nucleus neurons in the old animals. Since the inhibitory input to these neurons is mainly derived from within the suprachiasmatic nucleus, this reduction most likely reflects the greater proportion of silent cells found in old animals. The results show that the suprachiasmatic nucleus of old mice is subject to marked electrophysiological changes, which may contribute to physiological and behavioral changes associated with aging.

Galanin increases membrane excitability and enhances Ca2+ currents in adult, acutely dissociated dorsal root ganglion neurons

We examined the effect of galanin (10−15–10−7 m) on dispersed, mainly small‐sized dorsal root ganglion (DRG) neurons in adult rats using whole‐cell patch‐clamp. Galanin and AR‐M1896, a selective galanin type 2 receptor (GalR2) agonist, both significantly increased the number of action potentials in response to current pulses in 77% of the neurons, indicating an increase in excitability. Galanin also caused a rise in input resistance, decreased the holding current for −60 mV and depolarized the resting potential. In addition, Ca2+ currents elicited by voltage steps were significantly increased by both galanin and AR‐M1896 in nearly 70% of the cells. This enhancement was observed in 30% of the neurons in the presence of nimodipine or ω‐conotoxin, but in each case ≈ 60% less than without blocking either N‐ or L‐type Ca2+ channels, indicating modulation of both types of Ca2+ channels. The percentage of small‐ and medium‐sized neurons expressing GalR2 mRNA in DRGs in situ was similar to that showing increased excitability and Ca2+ current after galanin application, i.e. ≈ 70–80% of the neurons. The findings suggest that GalR2 has a role in controlling both the excitability, probably by inhibition of GIRK or leak K+ channels, and Ca2+ entry in a large population of presumably nociceptive neurons. The combination of the two effects, which possibly arise from separate biochemical pathways, would increase excitability and enhance intracellular Ca2+ signalling which would enhance sensory transmission. These mechanisms involving GalR2 receptors may underlie the pronociceptive effects of galanin described in the literature.

Disruption of Circadian Rhythms in Synaptic Activity of the Suprachiasmatic Nuclei by African Trypanosomes and Cytokines

Disturbances in biological rhythms pose a major disease problem, not the least in the aging population. Experimental sleeping sickness, caused by Trypanosoma brucei brucei, in rats constitutes a unique and robust chronic model for studying mechanisms of such disturbances. The spontaneous postsynaptic activity was recorded in slice preparations of the suprachiasmatic nuclei (SCN), which contain the master pacemaker for circadian rhythms in mammals, from trypanosome-infected rats. The excitatory synaptic events, which in normal rats show a daily variation, were reduced in frequency, while the inhibitory synaptic events did not significantly differ. This indicates selective disturbances in glutamate receptor-mediated neurotransmission in the SCN. Treatment with interferon-gamma in combination with lipopolysaccharide, which has synergistic actions with cytokines, and tumor necrosis factor-alpha similarly caused a reduction in excitatory synaptic SCN activity. We suggest that changes in the synaptic machinery of SCN neurons play an important pathogenetic role in sleeping sickness, and that proinflammatory cytokines can mimic these changes.

Interferon-γ-induced changes in synaptic activity and AMPA receptor clustering in hippocampal cultures

Extended release of interferon-gamma (IFN-gamma) in the nervous system during immunological and infectious conditions may trigger demyelinating disorders and cause disturbances in brain function. The aim of this study was to examine the effects of IFN-gamma on neuronal function in rat hippocampal cell cultures by using whole cell patch clamp analysis together with quantitative immunocytochemistry. Acute application of IFN-gamma to differentiated neurons in culture caused no immediate neurophysiological responses, but recordings after 48 h of incubation displayed an increase in frequency of AMPA receptor (AMPAR)-mediated spontaneous excitatory postsynaptic currents (EPSCs). Quantitative immunocytochemistry for the AMPAR subunit GluR1 showed no alteration in receptor clustering at this time point. However, prolonged treatment with IFN-gamma for 2 weeks resulted in a significant reduction in AMPAR clustering on dendrites but no marked differences in EPSC frequency between treated neurons and controls could be observed. On the other hand, treatment of hippocampal neurons for 4 weeks, instituted at an immature stage (1 day in culture), caused a significant reduction in spontaneous EPSC frequency. These neurons developed with no overt alterations in dendritic arborization or in the appearance of dendritic spines as visualized by alpha-actinin immunocytochemistry. Nonetheless, there was a marked reduction in AMPAR clustering on dendrites. These observations show that a key immunomodulatory molecule, IFN-gamma, can cause long-term modifications of synaptic activity and perturb glutamate receptor clustering.