Activity-dependent development of the vertebrate nervous system.
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Animals↗
Biomedical subjects
P G Nelson
Publications and source records attributed to P G Nelson.
Revised trauma score: a triage tool in the accident and emergency department.
We evaluated the Revised Trauma Score (RTS) for the rapid identification of severely injured patients on their arrival at the accident and emergency department. A total of 1407 consecutively injured patients admitted to the Accident and Emergency Department of the Royal Victoria Hospital, Belfast, had their RTSs calculated on arrival. A trauma team, consisting of experienced senior doctors, was summoned for all patients with an abnormal RTS of 11 or less. Each patient also had their Injury Severity Score (ISS) calculated later, and this was compared with their RTS on admission. It was found that 53 patients had an ISS of 16 or above, indicating severe injury. The RTS identified 42 of these. Of the remaining 11, five had incurred penetrating trauma and three spinal cord injuries. Clinical examination revealed actual or potentially serious injuries in all eight patients. However, serious injuries were not recognized initially in three patients due to physiological compensation and/or a short time lapse between injury and arrival at hospital. On admission 49 patients had an abnormal RTS, but their final ISS was less than 16. Forty had injuries warranting urgent resuscitation or a period of observation. Of the remaining nine patients, five had fractures with an elevated respiratory rate, presumably due to pain, and the other four had a depressed level of consciousness which could be accounted for by alcohol intoxication alone. Our results reinforce some well-known points. First, alcohol alone should not be assumed to be the only cause for a depressed level of consciousness in patients with head injuries. Second, systolic hypotension following trauma is an important sign of serious injury. We recommend the use of the RTS as an aid to junior doctors in the recognition of seriously injured patients in the accident and emergency department. Furthermore, the score should be recalculated at frequent intervals while the patient remains in the department.
Blood Pressure↗
General surgical manpower, Victoria, Australia.
To provide information for the planning of general surgical manpower in Victoria for the next 2 decades, an attempt was made to establish a statewide profile of surgical practices and an analysis of surgical activity to determine whether surgeons' and the community's needs were being met. Of 343 practising general surgeons, 280 furnished returns to a questionnaire--a return rate of 81.6%. Responses to the questionnaire reveal that the current surgeon-to-population ratio is optimal, although certain imbalances and concerns exist. Surgeons are fully occupied regardless of age, location of practice or type of practice, and they work long hours. Most surgeons regard themselves as not overworked and most are satisfied, although not all do the type of work they would prefer. Current surgical training programmes, however, appear inadequate for future needs.
Adult↗
Calcium, network activity, and the role of NMDA channels in synaptic plasticity in vitro.
Functionally effective neuronal circuits are constructed through a competitive process that requires patterned neuronal activity elicited by structured input from the environment. To explore the mechanisms of this activity-dependent synaptic restructuring, we have developed an in vitro preparation of mouse spinal cord neurons maintained in a 3-chambered cell-culture system. Sensory afferents that received chronic electrical stimulation for 3-5 d developed stronger synaptic connections than unstimulated afferents converging onto the same postsynaptic spinal cord neuron. Exposure to 100 microM DL-2-amino-5-phosphonovaleric acid (APV), an antagonist of the NMDA channel, during the stimulation period prevented the competitive advantage associated with electric stimulation. However, when APV was applied with a higher concentration of calcium (3 mM), activity-dependent synaptic plasticity was no longer inhibited by the NMDA receptor antagonist. This reversal of APV block of the plasticity was not impaired by reducing transmitter release with 3 mM magnesium (in addition to 3 mM calcium and APV). A suppressant effect of APV on spontaneous activity was observed, which was attributed to loss of the NMDA component of the EPSP. Activity-dependent plasticity was also blocked if spontaneous activity was suppressed with dilute tetrodotoxin (TTX; 5-10 nM), a dosage that reduces excitability of neurons but is insufficient to block sodium-dependent action potentials. These experiments bring into question how NMDA channel activation is involved in the processes of synaptic remodeling during development. The data suggest that postsynaptic activity is required for synaptic remodeling, but this activity need not involve NMDA receptor activation specifically for activity-evoked synaptic plasticity. Instead, the mechanism for plasticity appears to operate through calcium-dependent processes in general.
2-Amino-5-phosphonovalerate↗
N-methyl-D-aspartate receptors influence neuronal survival in developing spinal cord cultures.
Neuronal cell death, which exhibits precise spatial and temporal regulation, serves to remodel and optimize function in the developing nervous system. The mechanisms underlying neuronal cell death are poorly understood, but electrical activity and trophic substances appear to be among the important determinants of survival. We find that N-methyl-D-aspartate (NMDA) receptor antagonists induce neuronal cell death in developing spinal cord cultures. The magnitude of cell death is similar in amount to that produced by blocking action potentials with tetrodotoxin (TTX). The NMDA antagonists and TTX accelerate neuronal death in 2-week-old cultures but not in those that are 1 month old. Low concentrations of NMDA increased neuronal survival under conditions of electrical blockade with TTX. In addition, treatment with low levels of a calcium ionophore also decreased cell death associated with TTX. These results suggest that the NMDA receptor is an important determinant of neuronal survival and that this influence is stage-dependent and likely to be calcium-mediated.
2-Amino-5-phosphonovalerate↗
Cholinergic function in cultures of mouse spinal cord neurons.
Cholinergic synapses formed in cultures of fetal mouse spinal cord (SC) and superior cervical ganglion (SCG) were studied using intracellular and extracellular stimulation and recording as well as immunohistochemical staining for choline acetyltransferase (ChAT). Dissociated SC neurons and SC explants exhibited cholinergic terminals on SCG and SC neurons as demonstrated by ChAT immunoreactivity. Intracellular recordings showed that cholinergic inputs to SCG neurons were relatively common and that these synaptic inputs were blocked by the nicotinic acetylcholine (ACh) receptor blocker, tubocurarine. A comparison of three preparations indicated that the incidence of cholinergic activity recorded in SCG neurons was significantly higher in co-cultures of SCG with spinal cord ventral horn (VH) neurons grown on a substrate of non-neuronal cells from cerebral cortex, than in co-cultures with VH alone or with SC and dorsal root ganglion cells. Consistency between cholinergic physiology and staining for ChAT-positive terminals on SCG neuronal somata was obtained in cultures of SC explants grown with dissociated SCG. Application of acetylcholine, muscarine, and/or vasoactive intestinal polypeptide (VIP) produced slow excitation of SC neurons. Fast excitatory cholinergic interactions between SC neurons were not observed. Excitatory synaptic interactions between SC neurons were augmented by ACh or muscarine, while inhibitory synaptic interactions were diminished. Both types of synaptic modulation probably were produced by a presynaptic mechanism. Acetylcholine or muscarine affected synaptic interactions between SC neurons in only one-third of the synaptic connections tested, suggesting that the incidence of presynaptically cholinoceptive SC neurons is low in dissociated cell cultures. The experimental results show that a culture system incorporating dissociated fetal mouse SC neurons or explants of SC with sympathetic ganglion neurons expresses both nicotinic and muscarinic cholinergic function.
Acetylcholine↗
Mechanisms involved in activity-dependent synapse formation in mammalian central nervous system cell cultures.
Differences in neuronal activity produced by electrical stimulation lead to competition between synapses from sensory afferents converging on a common spinal cord neuron. Studies were performed on neurons dissociated from the mouse spinal cord and grown in culture dishes with three compartments. Synaptic efficacy from stimulated afferents was increased compared with unstimulated convergents, and the number of functional connections was increased by stimulation compared with control cultures. Blocking NMDA channel activation with 100 microM APV in medium containing 1.8 mM calcium inhibited this synaptic plasticity, but plasticity was not blocked by APV in medium in which the calcium concentration was elevated to 3 mM. These experiments support the view that electrical activity differentially influences processes that cause a persistent decrease in synaptic efficacy or lead to synapse elimination and those that increase synaptic strength or lead to synapse augmentation. We interpret our results in terms of a model in which these antagonistic mechanisms are both regulated via changes in calcium levels and second messengers that are modulated by electrical activity. A significant portion of the activity-related calcium influx relevant to synaptic plasticity passes through the NMDA channel, but other sources of calcium are involved. In particular, competitive elimination of synapses appears to occur during blockade of NMDA channels if the extracellular concentration of calcium is elevated moderately. The outcome of competition between the two calcium-dependent but antagonistic processes may depend either on their differential sensitivity to intracellular calcium concentration or separate specificities to NMDA and non-NMDA receptor-linked mechanisms.
Action Potentials↗
Multi-determinate regulation of neuronal survival: neuropeptides, excitatory amino acids and bioelectric activity.
Neuronal survival of dorsal root ganglion-spinal cord cultures was determined after treatment with vasoactive intestinal peptide (VIP) and an antagonist to the N-methyl-D-aspartate receptor (NMDA). Blockade of NMDA receptors with 2-amino 5-phosphonovaleric acid (AP5) produced a biphasic response on neuronal survival: low concentrations (0.1 microM) resulting in greater survival and higher concentrations (100 microM) causing cell death. VIP, a substance with demonstrated neurotrophic properties in vitro, prevented the neuronal cell death associated with high concentrations of AP5, while having no additive effect on the survival-promoting action of low levels of AP5. Electrophysiological studies indicated that AP5, although reducing high frequency bursting activity, did not significantly reduce the abundant on-going asynchronous activity present in these cultures of high density neuronal networks. These data indicate that excitatory amino acids have more than one action that can influence neuronal survival during development and that VIP can increase neuronal survival in bioelectrically active cultures when NMDA channels are blocked. Together with previous studies, these data suggest that multiple neurochemical inputs serve to determine the survival of spinal cord neurons during development, perhaps through one final common pathway: intracellular calcium regulation.
2-Amino-5-phosphonovalerate↗
Brains, behaviors, and magic bullets.
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Behavior↗
Effects of patterned electrical activity on neurite outgrowth from mouse sensory neurons.
A noninvasive method of electric stimulation was used in cell culture preparations to determine the effects of patterned electrical activity on the morphology and motility of mammalian central nervous system growth cones. Neurites from dorsal root ganglion (DRG) neurons of fetal mice were allowed to grow under the barrier of an insert placed in culture dishes. The insert confined the cell bodies within separate experimental and control compartments, and provided a means of exciting action potentials in the growing neurites by extracellular current pulses delivered across the barrier. A phasic pattern of stimulation caused immediate retraction of the filopodia and lamellipodium. Further outgrowth was halted and in many cases retraction of the neurite ensued. No changes in morphology or growth cone motility were evoked by electric stimulation when action potentials were blocked with 1 microM tetrodotoxin (TTX). These effects depended on the rate, pattern, and duration of stimulation. Phasic stimulation was more effective than stimulation with the same number of impulses delivered at a constant frequency. An important new observation was that cultures exposed to phasic stimulation for several hours contained actively growing neurites with normal growth cones which were insensitive to the stimulus. This apparent accommodation in neurites exposed to chronic stimulation may involve processes that regulate calcium conductance or buffering. Cessation of neurite outgrowth by action potentials could represent one mechanism linking morphological and functional characteristics in the developing CNS of mammals, by stabilizing the outgrowth of neurites forming appropriate synaptic contacts and leading to the retraction of growth cones from collaterals that have not formed appropriate contacts at the time the neuron enters into a functionally active circuit.
Action Potentials↗
Synaptic connections in vitro: modulation of number and efficacy by electrical activity.
The functional architecture of synaptic circuits is determined to a crucial degree by the patterns of electrical activity that occur during development. Studies with an in vitro preparation of mammalian sensory neurons projecting to ventral spinal cord neurons slow that electrical activity induces competitive processes that regulate synaptic efficacy so as to favor activated pathways over inactive convergent pathways. At the same time, electrical activity initiates noncompetitive processes that increase the number of axonal connections between these sensory and spinal cord neurons.
Action Potentials↗
The effect of intracellular tetraethylammonium ions on the reversal potential of monosynaptic EPSPs and excitatory amino acids in cultured spinal cord neurones.
Using the whole cell patch clamp recording technique, the reversal potential of currents generated by the application of excitatory amino acids and by stimulation of excitatory synapses has been examined in cultured spinal cord neurones, with the partial substitution of intracellular K+ by TEA+. A previous report (Clements et al. 1986) showed that intracellular injection of TEA into the soma of spinal motoneurones in the cat shifted the reversal of the 1a EPSC to more positive potentials, but the reason for this shift was not clear. The results reported here show that the observed change in the reversal potential for both excitatory amino acid agonists and the excitatory synaptic current, was not due to a change in the relative permeability of the channels to potassium or sodium, but could be explained by a change in driving force for the permeant intracellular ions (K+ and/or Cs+) on their substitution with the impermeant TEA+ ion.
Animals↗
Calcium agonist (BayK 8644) augments voltage-sensitive calcium currents but not synaptic transmission in cultured mouse spinal cord neurons.
The effects of the calcium agonist, BayK 8644, and other agents upon voltage-dependent calcium conductance (VSCC) and evoked synaptic activity were studied in cultured mouse spinal cord and dorsal root ganglion neurons. As expected, BayK 8644 increased the VSCC corresponding to L channels. It had relatively little effect on evoked synaptic activity; the small but statistically significant effect that was noted was a decrease. Nitrendipine had either no effect or an increase with no statistically significant effect being seen with regard to synaptic activity over the population sampled. An increased extracellular Ca++ concentration increased both VSCC and synaptic activity. We conclude that VSCC with L channel properties are probably not involved in transmitter release produced by action potentials in the central synapses occurring in the dissociated mouse spinal cord cell culture system.
3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
Differential effects of tetanus toxin on inhibitory and excitatory synaptic transmission in mammalian spinal cord neurons in culture: a presynaptic locus of action for tetanus toxin.
Tetanus toxin reduces monosynaptic inhibitory and excitatory synaptic transmission in mouse spinal cord neurons in culture. Inhibitory transmission is preferentially reduced by the toxin; however, excitatory transmission is also ultimately reduced and blocked by the concentrations of toxin used in these studies. Recordings from monosynaptically connected cell pairs revealed a marked diminution in amplitude of evoked monosynaptic inhibitory postsynaptic potentials coincident with the onset of convulsant action at a time when evoked monosynaptic EPSPs were relatively unaffected. Increased polysynaptic excitation occurred as a result of diminished inhibition. This supports the reduction of inhibition as an important mechanism in the convulsant action of tetanus toxin. Quantal analysis of the late effects of tetanus toxin on the monosynaptic excitatory postsynaptic potential revealed a reduction in quantal number with no reduction in quantal size, thus demonstrating a presynaptic locus of action for the toxin on spinal neurons.
Animals↗
Coordinate regulation of the expression of axonal proteins by the axonal microenvironment.
The axonal functions that act in the formation of the neuronal network have been shown to occur in close interdependence with the tissue that surrounds the growing axons. However, little is known about the molecular building blocks underlying axonal functions, although more than 400 axonal proteins have been identified. In view of the existence of such a large number of axonal proteins, we have initiated a project to determine the molecules involved in the implementation of particular axonal functions by a selective approach. On the assumption that plasticity in the expression of axonal functions in response to specific features of the local axonal environment may be based on changes in the expression of particular axonal proteins, the axonal proteins of dorsal root ganglion (DRG) neurons were screened for those whose expression responds to environmental influences. DRG neurons were grown in a compartmental cell system that offers separate access to neuronal somas and to their axons and the axons were locally exposed to different populations of cells from the peripheral or central nervous system. The axonal proteins were metabolically labeled and subjected to two-dimensional gel electrophoresis. Computerized quantitation of the individual axonal proteins revealed that the cocultured cells modulate the synthesis of a few axonal proteins of DRG neurons differentially. The data on the abundance of the newly expressed proteins under varying local environmental conditions were condensed as expression profiles. Comparison of expression profiles and cluster analysis of quantitative gel analysis data revealed that the environmentally modulated proteins subdivide into clusters with common distinct expression profiles under the influence of nonneuronal cells from the peripheral nervous system, nonneuronal cells of the central nervous system, and spinal cord cells, which are composed of neurons and nonneuronal cells. By means of this new, characteristic attribute assigned to environmentally modulated axonal proteins, working hypotheses were made as to their functional role.
Animals↗
Tetanus toxin in dissociated spinal cord cultures: long-term characterization of form and action.
The clinical course of tetanus is notable, in addition to its often dramatic clinical presentation, by the long duration of the neuromuscular symptoms. Survivors may have tetanic manifestations for several weeks after the onset of the disease. In this article we correlate the duration of specific electrophysiologic effects produced by tetanus toxin with the degradation of cell-associated toxin in primary cultures of mouse spinal cord neurons. From these studies we can conclude that the toxin has a half-life of 5-6 days. Both the heavy and the light chains of tetanus toxin degrade at similar rates. Labeled toxin, visualized by radioautography, is associated with neuronal cell bodies and neurites, and its distribution is not altered during a 1-week period following toxin exposure. Blockade of synaptic activity persists for weeks at the concentration of radiolabeled toxin used in these studies. This blockade of transmission is reversed as the toxin is degraded, suggesting that degradation of toxin may be a sufficient mechanism for recovery from tetanus.
Action Potentials↗
Synaptic excitation in cultures of mouse spinal cord neurones: receptor pharmacology and behaviour of synaptic currents.
Fast monosynaptic excitatory post-synaptic potentials between spinal cord neurones in cell culture (s.c.-s.c. e.p.s.p.s) were studied with current-clamp and two-electrode voltage-clamp methods. The reversal potential, response to acidic amino acid antagonists, and behaviour of the synaptic current were examined. The amplitude of the e.p.s.p. increased with membrane potential hyperpolarization and decreased with depolarization. The reversal potential of the e.p.s.p. was +3.8 +/- 2.5 mV (mean +/- S.E. of mean). The reversal potential for responses to ionophoretically applied L-glutamate and L-aspartate was also near 0 mV. The acidic amino acid antagonist, cis-2,3-piperidine dicarboxylic acid (PDA, 0.25-1.0 mM) reversibly antagonized the monosynaptic e.p.s.p.s as well as responses to kainate (KA) or quisqualate (QA). The selective N-methyl-D-aspartate antagonist, (+/-) 2-amino-5-phosphonovaleric acid (APV), had little effect on either the monosynaptic e.p.s.p.s or responses to QA or KA at concentrations that abolished responses to L-aspartate. Under voltage clamp, the peak synaptic current (e.p.s.c.) was linearly related to the membrane potential, increasing in amplitude with hyperpolarization and decreasing with depolarization from the resting potential. The decay of a somatic e.p.s.c. was well fitted by a single exponential function with a time constant of 0.6 ms at 25 degrees C. E.p.s.c.s which had proximal dendritic locations had decay time constants of 1-2 ms. The decay time constant was voltage-insensitive between -80 and +10 mV. We suggest that an acidic amino acid receptor other than that for NMDA mediates excitatory transmission at the s.c.-s.c. synapse; and that the underlying conductance mechanism is voltage insensitive with an estimated mean channel lifetime of less than 1 ms.
2-Amino-5-phosphonovalerate↗
Intracellular tetraethylammonium ions enhance group Ia excitatory post-synaptic potentials evoked in cat motoneurones.
Single fibre group Ia excitatory post-synaptic potentials (e.p.s.p.s) were recorded in cat spinal motoneurones after the neurones were injected with tetraethylammonium (TEA) ions. TEA injection increased the peak amplitude of most e.p.s.p.s. The time course of e.p.s.p.s generated at the soma was unaffected, but the time course of e.p.s.p.s generated in the dendrites was prolonged. The membrane time constant did not change after TEA injection. Somatic e.p.s.p.s were voltage clamped after TEA was injected. The reversal potential for these e.p.s.p.s was more positive than for e.p.s.p.s unaffected by TEA. Composite e.p.s.p.s added linearly, or greater than linearly, whereas in motoneurones without TEA they added linearly or less than linearly. The enhanced amplitude and prolonged time course observed in dendritic e.p.s.p.s after TEA injection was reduced by small hyperpolarizing currents. Greater than linear summation of composite e.p.s.p.s was converted to linear summation by small hyperpolarizing currents. The increase in somatic e.p.s.p.s was attributed to a more positive reversal potential for the e.p.s.p.s. We suggest that TEA decreases the relative permeability of K+ in the subsynaptic channels. We propose that in the presence of TEA, dendritic depolarization activates an inward current which amplifies and prolongs synaptic potentials spreading towards the soma.
Action Potentials↗