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C J Woolf

Publications and source records attributed to C J Woolf.

At least 109 records · Page 6Linked to original sources
PubMed1993

The role of neurokinin and N-methyl-D-aspartate receptors in synaptic transmission from capsaicin-sensitive primary afferents in the rat spinal cord in vitro.

The rat spinal cord with connected dorsal root ganglia was used to study neurokinin and N-methyl-D-aspartate receptors involved in the sensory synaptic transmission of dorsal horn cells. Selective C-fibre excitation was produced by capsaicin (200-500 nM) administered to the dorsal root ganglions. Sixty-nine per cent of dorsal horn cells responded with a postsynaptic depolarization and enhanced synaptic activity, recorded via intracellular electrodes, to capsaicin-activated primary afferent input. Dorsal horn neurons activated by the capsaicin-evoked input were also excited by a 1-min perfusion of the neurokinin-1 receptor agonists substance P methyl ester or GR73 632 and by the neurokinin-2 agonist neurokinin-A. These cells were also depolarized by N-methyl-D-aspartate. Responses to substance P methyl ester and GR73 632 were selectively reduced by the neurokinin-1 receptor antagonist CP96,345, and responses to neurokinin-A were completely blocked by the neurokinin-2 receptor antagonist MEN10 376. The depolarization evoked by N-methyl-D-aspartate was not altered by either of the antagonists, but was completely blocked by the selective N-methyl-D-aspartate receptor antagonist (-)-2-amino-5-phosphonovaleric acid. Capsaicin-evoked responses in the dorsal horn were inhibited by MEN10,376 (63 +/- 13% inhibition) but no significant change was observed with CP96,345. The N-methyl-D-aspartate receptor antagonist (-)-2-amino-5-phosphonovaleric acid consistently inhibited the capsaicin-induced response by 76 +/- 14%. Combination of (-)-2-amino-5-phosphonovaleric acid and MEN10,376 produced an almost complete abolition of the capsaicin-evoked depolarization.(ABSTRACT TRUNCATED AT 250 WORDS)

2-Amino-5-phosphonovalerate↗
PubMed1993

Cobalt uptake enables identification of capsaicin- and bradykinin-sensitive subpopulations of rat dorsal root ganglion cells in vitro.

A novel modification of the stimulated cobalt uptake technique has been used to identify rat dorsal root ganglion cells expressing capsaicin and bradykinin receptors. The technique involves incubating intact dorsal root ganglia in vitro in a modified Krebs solution in which cobalt chloride has been substituted for calcium. Activation of dorsal root ganglion cells by capsaicin or bradykinin in the presence of the cobalt ions results in cobalt influx into the excited cells. Histochemical methods were then used to visualize the intracellular accumulation of cobalt, and labelled cells were counted and characterized. Capsaicin (2 microM) or bradykinin (500 nM) applied for 20 min induced cobalt uptake in 13.8 +/- 0.6 and 9.6 +/- 0.5% of neuronal profiles in dorsal root ganglia (L4), respectively, a significantly larger number than stained in control ganglia (in the absence of agonists: 1.8 +/- 0.7%). The longest diameter of the soma of stained dorsal root ganglion cells following capsaicin and bradykinin perfusion were significantly different from each other and from the non-labelled population (17.5 +/- 0.7 and 24.5 +/- 0.2 microns for capsaicin; 23.2 +/- 0.9 and 25.5 +/- 0.4 microns for bradykinin; labelled and non-labelled cells, respectively). The distribution of cell diameters revealed that while capsaicin-sensitive cells were exclusively small-sized, bradykinin-sensitive cells were predominantly small and medium sized. The selective bradykinin-2 receptor antagonist HOE-140 (5.0 microM) blocked the bradykinin-induced staining (2.16 +/- 0.02%) but not that of capsaicin. The bradykinin-1 agonist [des-Arg9]-bradykinin did not induce any significant increase in stained cells over the control number (2.2 +/- 0.7%).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Reciprocal Schwann cell-axon interactions.

This article describes the reciprocal interactions between neurones and Schwann cells with particular reference to the role of growth factors and neurokines as signalling molecules between these cells and of the extracellular matrix as a conduit for such signalling. Major recent advances have identified molecules produced by neurones that are responsible for Schwann cell proliferation, as well as some of the Schwann cell factors regulating the expression of molecules shown to play an important role in neuronal survival and differentiation.

Animals↗
PubMed1993

Rate of rise of the cumulative depolarization evoked by repetitive stimulation of small-caliber afferents is a predictor of action potential windup in rat spinal neurons in vitro.

1. The summation of depolarizing postsynaptic potentials (PSPs) evoked by stimulation of primary afferent fibers in lumbar dorsal roots was studied in dorsal and ventral horn neurons in the rat hemisected spinal cord in vitro with current-clamp intracellular recording techniques. PSPs evoked by activation of A delta and/or C fibers could summate after repetitive stimulation at low frequencies: this resulted in a progressive, long-lasting change in the neuronal membrane potential (Vm) (cumulative depolarization). Cumulative depolarization was not observed after stimulation at A beta-fiber intensity or at frequencies < 0.2 Hz. 2. Effective PSP summation resulting in a significant cumulative depolarization (> 5 mV) was observed only if the PSPs evoked were longer than 4 s. On the other hand, there was no correlation between the amplitude of cumulative depolarization achieved in a neuron and the absolute duration of the PSP or the neuronal resting membrane properties (Vm and input resistance). 3. With relation to the changes in Vm during the stimulation train, three patterns of response to repetitive stimuli could be detected in both dorsal horn (DH) and ventral horn (VH) neurons: 1) little or no depolarization throughout the train; 2) an initial depolarization, which either remained stationary or decayed to the resting Vm after the first 3-4 s of stimulation; or 3) a progressive cumulative depolarization increasing throughout the train. 4. Depolarizing the cell by DC current injection resulted in a steeper rise in Vm in response to repetitive stimuli in one half of VH neurons and in one out of nine DH neurons tested. 5. Action potential windup, an increase in the number of action potentials elicited by each stimulus as the train progresses, was observed both in DH and in VH cells. C-fiber strength stimulation was the most effective in eliciting windup. Windup was associated with Vm responses to repetitive stimulation belonging to the progressive cumulative depolarization pattern. Indeed, cumulative depolarizations that resulted in windup had significantly steeper slopes for the Vm trajectory than those that did not. In those neurons in which DC depolarization by intracellular current injection increased the slope of the Vm trajectory, DC depolarization could also bring about action potential windup. 6. These results indicate that, although the generation of long-lasting "slow" PSPs by high-threshold primary afferents is necessary for the summation of synaptic activity in the spinal cord at low frequencies, the rate and pattern of the summation does not depend on the absolute duration of the PSPs.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗
PubMed1993

Small-caliber afferent inputs produce a heterosynaptic facilitation of the synaptic responses evoked by primary afferent A-fibers in the neonatal rat spinal cord in vitro.

1. The effect of brief primary afferent inputs on the amplitude and duration of the synaptic potentials evoked in ventral horn (VH) neurons by the activation of other unconditioned primary afferents was studied by current-clamp intracellular recording in the neonatal rat hemisected spinal cord in vitro. Low-frequency (1 Hz) trains of stimulation were applied to a lumbar dorsal root (Conditioning root) for 20-30 s. Test excitatory synaptic potentials (EPSPs) were evoked by single electrical shocks applied to an adjacent Test dorsal root. 2. Test and Conditioning inputs were generated at stimulation strengths sufficient to activate A beta-, A delta- and C-afferent fibers successively. At A delta- and C-fiber strength the EPSPs lasted for 4-6 s, and, during the repetitive Conditioning inputs, these summated to produce a progressively incrementing cumulative depolarization that slowly decayed back to the control Vm over tens of seconds. 3. Dorsal root conditioning produced heterosynaptic facilitation, defined as an enhancement of Test EPSPs above their DC matched controls, in 7 out of 20 neurons. To facilitate the unconditioned afferent input, the intensity of conditioning stimulation had to exceed the threshold for the activation of thin myelinated (A delta) afferents: conditioning at A beta-fiber strength had no effect, whereas A delta- and C-fiber strength conditioning were equally effective. 4. Heterosynaptic facilitation of only A beta- or A delta-fiber-evoked Test EPSPs was observed, no enhancement of C-fiber strength Test EPSPs could be demonstrated. The facilitation manifested as increases in the EPSP peak amplitude, area or the number of action potentials evoked. 5. Conditioning trials that produced heterosynaptic facilitation generated cumulative depolarizations larger than those produced by ineffective conditioning trials (9.1 +/- 3.1 vs. 3.3 +/- 0.5 mV after 20 s conditioning at resting Vm, mean +/- SE, n = 6 and 13, respectively; P < 0.05). The slope of the Vm trajectory during the summation of the conditioning EPSPs was higher in trials resulting in heterosynaptic facilitation, at 0.31 +/- 0.10 mV/s in neurons with heterosynaptic facilitation and 0.06 +/- 0.02 mV/s in cells without heterosynaptic facilitation (P < 0.05). 5. Four of the 20 VH neurons in our sample responded to A delta/C-fiber conditioning with action-potential windup: all 4 also displayed heterosynaptic facilitation. 6. Heterosynaptic facilitation decayed after the completion of the conditioning stimulus with a time course that was parallel to but not superimposable on that of the slow Vm depolarization evoked by the conditioning.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1992

Long duration ventral root potentials in the neonatal rat spinal cord in vitro; the effects of ionotropic and metabotropic excitatory amino acid receptor antagonists.

Long duration, primary afferent evoked ventral root potentials (VRP's) have been recorded in vitro from hemisected spinal cords prepared from 8-12-day-old rat pups. Single shock stimulation of a dorsal root at stimulus strengths sufficient to recruit C/group IV afferent fibres evoked a long duration (11.9 +/- 1.2 s) ipsilateral VRP in all preparations. This long duration VRP consisted of two components, (i) a slow wave, time to peak 137.0 +/- 5.1 ms, the amplitude of which was reduced to 8.7% of mean control value in the presence of the N-methyl-D-aspartate (NMDA) antagonist D-AP5 (40 microM), (ii) a prolonged wave with a time to peak of 2.0 +/- 0.2 s which was partially resistant to D-AP5 (40 microM). Both the slow and the prolonged waves were unaffected following superfusion with the metabotropic excitatory amino acid (EAA) receptor antagonist L-AP3 (100-200 microM). Low frequency (1-10 Hz) repetitive stimulation (20 s duration) of high threshold dorsal root afferents evoked a temporal summation of synaptic activity which generated a progressively depolarizing VRP. This cumulative VRP was graded with frequency of stimulation (0.89 +/- 0.13 to 1.25 +/- 0.19 mV). The cumulative VRP was followed by a post-stimulus depolarization which outlasted the period of repetitive stimulation by tens of seconds (47.6 +/- 8.4 to 91.2 +/- 19.9 s). In the presence of AP5 the amplitude of the cumulative VRP was depressed to 54.5 +/- 11.5% of control values when low frequency (1.0 Hz) stimulation was used. The proportion of the cumulative VRP resistant to D-AP5 increased as the frequency of stimulation was increased to 10 Hz. The decay time of the post-stimulus depolarization was unaffected by AP5. Neither the amplitude nor the post-stimulus depolarization of the cumulative VRP was affected by 200 microM L-AP3. It is suggested that both an AP5 sensitive and AP5 insensitive potential contribute to the long duration VRP evoked in the neonatal rat spinal cord following single shock high threshold afferent stimulation. Moreover, the AP5 insensitive prolonged depolarization is manifest following sustained low frequency stimuli and higher frequency inputs.

2-Amino-5-phosphonovalerate↗
PubMed1992

Regional differences in the distribution of capsaicin-sensitive target-identified adult rat dorsal root ganglion neurons.

A retrograde labelling technique combined with a cobalt uptake assay in cultured adult rat dorsal root ganglion (DRG) neurons were applied to study the distribution of capsaicin sensitivity in relation to different peripheral targets. This study shows that there are regional differences between skin, skeletal muscle and urinary bladder; 20-30% of skin afferents, 40% of muscle afferents and 60% of bladder afferents were found to be capsaicin-senditive. This may reflect differences in the proportion of chemosensitive afferents innervating different peripheral tissues.

Animals↗
PubMed1992

Peripheral nerve injury triggers central sprouting of myelinated afferents.

The central terminals of primary afferent neurons are topographically highly ordered in the spinal cord. Peripheral receptor sensitivity is reflected by dorsal horn laminar location: low-threshold mechanoreceptors terminate in laminae III and IV (refs 2, 3) and high-threshold nociceptors in laminae I, II and V (refs 4,5). Unmyelinated C fibres, most of which are nociceptors, terminate predominantly in lamina II (refs 5, 7). There is therefore an anatomical framework for the transfer of specific inputs to localized subsets of dorsal horn neurons. This specificity must contribute to the relationship between a low-intensity stimulus and an innocuous sensation and a noxious stimulus and pain. We now show that after peripheral nerve injury the central terminals of axotomized myelinated afferents, including the large A beta fibres, sprout into lamina II. This structural reorganization in the adult central nervous system may contribute to the development of the pain mediated by A-fibres that can follow nerve lesions in humans.

Afferent Pathways↗
PubMed1992

Terminal Schwann cells elaborate extensive processes following denervation of the motor endplate.

Terminal Schwann cells, when stained for S100 (a calcium binding protein), can be seen to cap motor axons at the neuromuscular junction. Within days of denervation the Schwann cells begin to stain for the low affinity nerve growth factor receptor, but remain Thy-1 negative, and elaborate fine processes. These processes become longer and more disorganized over weeks, and cells positive for S100 and nerve growth factor receptor migrate into the perisynaptic area. Reinnervation results in a withdrawal of the processes. The morphology and location of terminal Schwann cells seems to depend on axonal contact. The spread of Schwann cells and their processes away from the synaptic zone following denervation, implies that these cells do not target axons directly to the endplate.

Animals↗
PubMed1992

GAP-43 mRNA in Rat Spinal Cord and Dorsal Root Ganglia Neurons: Developmental Changes and Re-expression Following Peripheral Nerve Injury.

The expression of growth-associated protein GAP-43 mRNA in spinal cord and dorsal root ganglion (DRG) neurons has been studied using an enzyme linked in situ hybridization technique in neonatal and adult rats. High levels of GAP-43 mRNA are present at birth in the majority of spinal cord neurons and in all dorsal root ganglion cells. This persists until postnatal day 7 and then declines progressively to near adult levels (with low levels of mRNA in spinal cord motor neurons and 2000 - 3000 DRG cells expressing high levels) at postnatal day 21. A re-expression of GAP-43 mRNA in adult rats is apparent, both in sciatic motor neurons and the majority of L4 and L5 dorsal root ganglion cells, 1 day after sciatic nerve section. High levels of the GAP-43 mRNA in the axotomized spinal motor neurons persist for at least 2 weeks but decline 5 weeks after sciatic nerve section, with the mRNA virtually undetectable after 10 weeks. The initial changes after sciatic nerve crush are similar, but by 5 weeks GAP-43 mRNA in the sciatic motor neurons has declined to control levels. In DRG cells, after both sciatic nerve section or crush, GAP-43 mRNA re-expression persists much longer than in motor neurons. There was no re-expression of GAP-43 mRNA in the dorsal horn of the spinal cord after peripheral nerve lesions. Our study demonstrates a similar developmental regulation in spinal cord and DRG neurons of GAP-43 mRNA. We show moreover that failure of re-innervation does not result in a maintenance of GAP-43 mRNA in axotomized motor neurons.

Journal Article↗
PubMed1992

Denervation of the motor endplate results in the rapid expression by terminal Schwann cells of the growth-associated protein GAP-43.

Developing and regenerating neurons express high levels of the growth-associated phosphoprotein GAP-43. This membrane protein is not confined to neurons, however, as a number of studies have demonstrated GAP-43 immunoreactivity in central and peripheral glia in vitro and in vivo. We have found that the Schwann cells overlying the terminal motor axon at adult rat skeletal muscle endplates, and the motor axons themselves, are normally not GAP-43 immunoreactive. Within 24 hr of denervation, however, the terminal Schwann cells are positive for a GAP-43 mRNA in situ hybridization signal and are GAP-43 immunoreactive. The immunoreactive GAP-43 cells possess elaborate processes that branch from the endplate region into the perisynaptic zone and stain with defined Schwann cell markers: the calcium binding protein S100 and the low-affinity NGF receptor (NGFr), but not with a fibroblast marker, Thy-1. Reinnervating motor axons are GAP-43 positive, with an appearance quite different from the GAP-43-positive Schwann cells. The reappearance of nerve endings at the motor endplate is followed by the disappearance of GAP-43 labeling in the Schwann cells and of a retraction of their processes. GAP-43 expression in Schwann cells is therefore state dependent, apparently regulated by neural contact. This protein, which is associated in neurons with neurite formation, may participate in the elaboration of processes by Schwann cells when their contact with axons is disrupted.

Animals↗
PubMed1991

Distribution of the growth associated protein GAP-43 in the central processes of axotomized primary afferents in the adult rat spinal cord; presence of growth cone-like structures.

GAP-43-immunolabelled structures were visualized by electron microscopy in the adult rat L4-L5 superficial dorsal horn 2 weeks after sciatic nerve transection. The majority of immunolabelled elements were unmyelinated axons, but some synaptic terminals and myelinated axons also labelled. The labelled unmyelinated axons were commonly located in prominent bundles which on serial section analysis could be followed into larger single trunks. These enlargements contain many organelles and give rise to smaller processes, which is compatible with their being growth cones. Sciatic nerve transection may result, therefore, in central regenerative processes which reorganize the neuropil and contribute to the decreased sensibility and pain that follows peripheral nerve section.

Afferent Pathways↗
PubMed1991

Common patterns of plasticity contributing to nociceptive sensitization in mammals and Aplysia.

In contrast to innocuous stimuli, which only have transient effects when applied to the body surface, noxious stimuli generate persistent changes in the nervous system. This nociceptive memory manifests itself most prominently as a post-injury sensitization where, after tissue damage, the avoidance reaction and pain that result from subsequent stimuli are exaggerated and prolonged and can be initiated by low intensity stimuli. Similarities between nociceptive sensitization in mammals (including humans) and the mollusc Aplysia californica suggest that fundamental mechanisms contributing to injury-induced behavioral modifications might be widespread in the animal kingdom.

Animals↗
PubMed1991

Time-dependent differences in the increase in GAP-43 expression in dorsal root ganglion cells after peripheral axotomy.

Peripheral axotomy of primary afferent neurons results in the up-regulation of the growth-associated phosphoprotein GAP-43, by dorsal root ganglion cells. We have studied the temporal sequence of GAP-43 expression in those dorsal root ganglion neurons with unmyelinated axons (the small dark cells) and in those with myelinated axons (the large light cells) after sciatic nerve section in the adult rat. Immunoreactivity for the RT 97 neurofilament epitope, which is detectable only in large light dorsal root ganglion cells, was used to differentiate the two types of dorsal root ganglion cell. Within two days of a sciatic nerve section the number of GAP-43-immunoreactive profiles in the ipsilateral ganglion had increased five-fold and this increase persisted for 80 days post-section. While 50% of the small numbers of GAP-43-positive cells in control ganglia were RT 97 positive, only 8% of the large number of GAP-43-immunoreactive cells four days post-section, were RT 97 positive. By 14 days the number of RT 97-positive/GAP-43-positive cells had increased to 29%. This was paralleled by an increase in GAP-43 immunoreactivity in large diameter profiles at 14 days. The signals that alter GAP-43 expression in unmyelinated (small, RT 97 -ve) and myelinated (large, RT 97 +ve) afferents after peripheral nerve injury appear to operate with different time-courses.

Animals↗
PubMed1991

Generation of acute pain: central mechanisms.

Pain can either be 'nociceptor-mediated', produced as a consequence of the activation of high threshold nociceptors, or 'A-fibre mediated', resulting from the activation of low threshold A beta afferent fibres. Under normal circumstances nociceptor mediated pain only occurs in response to high intensity noxious stimuli. Following peripheral tissue injury the inflammatory reaction generates a complex set of chemical signals that alter the transduction properties of nociceptors such that they can be activated by low intensity stimuli, the phenomenon of peripheral sensitization. Pain in this circumstance is still nociceptor mediated but can be generated by low intensity or innocuous stimuli. The nociceptive input to the spinal cord in these circumstances however produces activity-dependent alterations in the response properties of neurones in the dorsal horn. This means that they begin to respond to normal inputs, including that generated by A beta low threshold afferents, in an abnormal and exaggerated way. This is the phenomenon of central sensitization. Because afferent inputs can provoke prolonged alterations within the central nervous system, optimal treatment of acute pain states should be directed both at abolishing peripheral sensitization and to preventing the establishment of central sensitization. The latter involves the strategy of pre-emptive analgesia.

Afferent Pathways↗
PubMed1990

Collateral sprouting of the central terminals of cutaneous primary afferent neurons in the rat spinal cord: pattern, morphology, and influence of targets.

The capacity of the central terminals of primary afferents to sprout into denervated areas of neonatal spinal cord and the morphology of any novel terminals has been investigated. In rats which had undergone sciatic nerve section on the day of birth, 12 of 18 physiologically characterized intact saphenous hair follicle afferents (HFAs) were labelled intra-axonally with horseradish peroxidase (HRP) were shown to sprout up to 2,000 microns into the deafferented sciatic terminal field. The morphology of these sprouts depended on which area of the sciatic nerve territory was invaded by the afferent sprouts. Six HFAs sprouted into areas normally innervated by glabrous skin afferents and the morphology of the collateral sprouts in this region resembled that of rapidly adapting (RA) afferents. The other six saphenous HFAs had sprouted into sciatic "hairy" skin areas and the morphology of these sprouts, although abnormal, was flame shaped. In rats whose sural, saphenous, and superficial peroneal nerves were cut at birth, 4 of 7 single HRP labelled RA afferents had central terminals that had sprouted into regions of cord normally devoted to "hairy" input. These showed clear signs of HFA morphology despite their peripheral receptive fields remaining in the glabrous skin. The results show collateral sprouting of single cutaneous sensory afferent axons into adjacent inappropriate central target regions following neonatal deafferentation. Such plasticity may provide some compensation following neonatal injury. The morphology of the sprouted terminals is appropriate to the new target area rather than to its functional class and is also independent of the peripheral receptive field location providing an example of central rather than peripheral control over afferent growth patterns.

Afferent Pathways↗
PubMed1990

Neonatal capsaicin treatment induces invasion of the substantia gelatinosa by the terminal arborizations of hair follicle afferents in the rat dorsal horn.

Capsaicin, administered on the day of birth, was found to alter laminar distribution, but not the receptive field properties or the morphology of the collateral arborizations of hair follicle afferents (HFAs) intra-axonally injected with horseradish peroxidase (HRP). Of the 65 HFA terminal arbors in capsaicin treated rats, 46 (71%) were found to enter the substantia gelatinosa (in control rats, 44/165, 27%). All of the collaterals projected to somatotopically normal areas of cord. Dorsal horn shrinkage (21%), as estimated by planimetric measurements of Nissl and acetylcholinesterase-stained material, was only a partial explanation of this result. This idea was supported by the statistically significant increase (27%, P less than 0.05) in the absolute dorsoventral length of collaterals. The results show that the destruction of unmyelinated fibres during the early postnatal period by capsaicin induces HFA invasion into the area that C fibres normally occupy. This invasion suggests that the laminar termination sites for different primary afferent fibres are not altogether specified and that intact neonatal primary afferents have the capacity to sprout into denervated regions of spinal cord.

Animals↗