Low- and high-voltage-activated calcium currents in rat spinal dorsal horn neurons.

1. Calcium currents in immature rat spinal dorsal horn neurons in transverse slices were studied with the single-electrode voltage-clamp technique. Using experimental conditions that minimized voltage-dependent Na+ and K+ currents, we distinguished low- and high-voltage-activated calcium currents on the basis of their voltage dependence and sensitivity to the Ca2(+)-channel agonist and antagonist drugs. 2. The low-voltage-activated transient calcium current is evoked with weak depolarizing voltage commands. It begins to activate at potentials positive to -70 mV and increases in amplitude and rate of decay with depolarization, the peak values being reached between -40 and -30 mV. The current is fully activated at a holding potential of about -110 mV. Steady-state inactivation is complete at potentials in the range of -60 to -50 mV. 3. The transient component of the high-threshold calcium current appears at membrane potentials close to -40 mV and slowly decays within several hundreds of milliseconds. The amplitude of the current increases with more negative holding potentials (-100 to -40 mV). 4. The sustained component of the high-threshold calcium current seems to activate at potentials positive to -40 mV and exhibits little inactivation during 0.3- to 0.5-s depolarizing commands. This component is better isolated at more depolarized holding potentials (between -40 and -30 mV) that inactivate the transient components of the low- and high-threshold calcium currents. 5. A rundown of calcium currents was seen in dorsal horn cells. The time stability of the transient and sustained components of the high-threshold calcium current was lower than that of the low-threshold transient current. The latter current seemed to be insensitive up to 1 h. 6. (-)-Bay K 8644 (1-10 microM), a dihydropyridine agonist, enhanced the high-threshold calcium current, in particular the sustained component, but not the transient low-threshold calcium current. The dihydropyridine antagonist nifedipine (5-50 microM) selectively reduced the sustained component of the high-threshold calcium current while having little or no effect on the transient components of the low- and high-threshold calcium currents. 7. Cadmium ions (60-100 microM) and cobalt ions (2 mM) markedly reduced both components of the high-threshold calcium current, and Cd2+ only slightly decreased the low-threshold transient current. However, all three components are indiscriminately blocked by higher concentrations of Cd2+ and Co2+.(ABSTRACT TRUNCATED AT 400 WORDS)     

Thiopental inhibits glycine receptor function in acutely dissociated rat spinal dorsal horn neurons

Whole-cell patch-clamp was used to assess the modulatory effect of thiopental (Thio) on glycine (Gly) receptor in mechanically dissociated rat spinal dorsal horn neurons. It was found that Thio inhibited the amplitude, accelerated the desensitization and prolonged the deactivation of Gly-induced currents (I_Gly) in a concentration-dependent manner. In addition, a rebound current occurred after washout of the co-application of Gly and Thio in most neurons tested. Moreover, the inhibitory effect of Thio was not the result of cross-inhibition between Gly and GABA_A receptors. Furthermore, taurine-induced currents, a low-affinity agonist for Gly receptors, were also markedly inhibited by Thio in a similar way to I_Gly. These results indicate that Thio suppresses Gly receptor function and suggest that Thio anesthetic actions might not be mediated by Gly receptors. We speculate that the weak muscle relaxation and the limited analgesic effects observed during Thio anesthesia may attribute to its inhibitory effects on Gly receptors.     

The effect of serotonin on GABA synthesis in cultured rat spinal dorsal horn neurons

The spinal dorsal horn (SDH) is the first step in the integration of primary nociceptive information, which is controlled by the descending serotonin (5-HT) system as well as the principal inhibitory neurotransmitter gamma-aminobutyric acid (GABA). However, the influence exerted by 5-HT on GABA synthesis remains poorly understood. The major pathway for GABA synthesis is the enzymatic decarboxylation of glutamate by glutamic acid decarboxylase (GAD) 65 and 67. In the present research, western blotting results show a time- and dose-dependent enhancement of GAD65 and GAD67 expression induced by 5-HT treatment and a concentration of 100nM 5-HT applied for 3 days is shown to be the optimal condition for maximal expression of GAD67 and a significant expression of GAD65. Under the stimulation of such 5-HT application the phosphorylation of Akt and p42/p44 mitogen-activated protein (MAP) kinase is activated and specifically blocked by inhibitors of phosphatidylinositol 3-kinase (PI3-K) (LY294002) or the p42/p44 MAP kinase (PD98059 and U0126) pathways. Moreover, LY294002, or PD98059, or U0126 partially inhibit 5-HT-stimulated increases in GAD67 or GAD65 expression. Further, 5-HT application has no effect on the number of GAD65/GAD67-immunopositive neuronal cells; but it can induce an increase in the total area, process length and number of primary neurites of GAD65/67-positive neurons, an increase that appears to involve LY294002 and PD98059. The results of this study provide an in vitro model of the regulation of 5-HT on synthesis of GABA in the SDH that is putatively thought to occur in vivo as a result of excitatory neural activity.     

Inflammation-induced increase in evoked calcium transients in subpopulations of rat dorsal root ganglion neurons

The concentration of intracellular Ca²⁺ ([Ca²⁺]_i) influences neuronal properties ranging from excitability to neurotransmitter release. Persistent inflammation is associated with changes in the properties of primary afferent neurons ranging from excitability to transmitter release. The purpose of the present study was to determine whether previously described inflammation-induced changes in excitability and transmitter release are associated with changes in the regulation of [Ca²⁺]_i. Acutely dissociated dorsal root ganglion (DRG) neurons harvested from adult rats 3 days following a hind-paw injection of complete Freund's adjuvant (CFA) or naïve controls, were stimulated with 30 mM K⁺ (High K⁺). High K⁺ evoked changes in [Ca²⁺]_i were assessed with fura-2 ratiometric microfluorimetry. Subpopulations of DRG neurons were defined by cell body diameter, isolectin B4 (IB4) binding, capsaicin (CAP) sensitivity and target of innervation (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbo-cyanine perchlorate labeling). Inflammation was associated with significant increases in resting [Ca²⁺]_i and increases in the magnitude and decreases in the decay, of the evoked increase in [Ca²⁺]_i. The changes in evoked transients were larger in neurons innervating the site of inflammation. Furthermore, there were differences among subpopulations of DRG neurons with respect to changes in magnitude and/or decay of the evoked transient such that the increase in magnitude was larger in small- and medium-diameter neurons than in large diameter neurons while the decrease in the decay was greater in CAP responsive, IB4 positive, small- and medium-diameter neurons than in CAP unresponsive, IB4 negative and/or large-diameter neurons. These changes in the regulation of [Ca²⁺]_i were not due to inflammation-induced changes in passive or active electrophysiological properties. Importantly, an inflammation-induced increase in evoked Ca²⁺ transients in putative nociceptive afferents may contribute to the pain and hyperalgesia associated with persistent inflammation via facilitation of transmitter release from these afferents.     

Effect of streptozotocin-induced diabetes on the activity of calcium channels in rat dorsal horn neurons

We have previously found that spinal dorsal horn neurons from streptozotocin-diabetic rats, an animal model for diabetes mellitus, show the prominent changes in the mechanisms responsible for [Ca2+]i regulation. The present study aimed to further characterize the effects of streptozotocin-induced diabetes on neuronal calcium homeostasis. The cytoplasmic Ca2+ concentration ([Ca2+]i) was measured in Fura-2AM-loaded dorsal horn neurons from acutely isolated spinal cord slices using fluorescence technique. We studied Ca2+ entry through plasmalemmal Ca2+ channels during potassium (50 mM KCl)-induced depolarization. The K+-induced [Ca2+]i elevation was inhibited to a different extent by nickel ions, nifedipine and omega-conotoxin suggesting the co-expression of different subtypes of plasmalemmal voltage-gated Ca2+ channels. The suppression of [Ca2+]i transients by Ni2+ (50 microM) was the same in control and diabetic neurons. On the other hand, inhibition of [Ca2+]i transients by nifedipine (50 microM) and omega-conotoxin (1 microM) was much greater in diabetic neurons compared with normal animals. These data suggest that under diabetic conditions the activity of N- and L- but not T-type voltage-gated Ca2+ channels substantially increased in dorsal horn neurons.     

Effect of streptozotocin-induced diabetes on the activity of calcium channels in rat dorsal horn neurons

We have previously found that spinal dorsal horn neurons from streptozotocin-diabetic rats, an animal model for diabetes mellitus, show the prominent changes in the mechanisms responsible for [Ca²⁺]_i regulation. The present study aimed to further characterize the effects of streptozotocin-induced diabetes on neuronal calcium homeostasis. The cytoplasmic Ca²⁺ concentration ([Ca²⁺]_i) was measured in Fura-2AM-loaded dorsal horn neurons from acutely isolated spinal cord slices using fluorescence technique. We studied Ca²⁺ entry through plasmalemmal Ca²⁺ channels during potassium (50 mM KCl)-induced depolarization. The K⁺-induced [Ca²⁺]_i elevation was inhibited to a different extent by nickel ions, nifedipine and ω-conotoxin suggesting the co-expression of different subtypes of plasmalemmal voltage-gated Ca²⁺ channels. The suppression of [Ca²⁺]_i transients by Ni²⁺ (50 μM) was the same in control and diabetic neurons. On the other hand, inhibition of [Ca²⁺]_i transients by nifedipine (50 μM) and ω-conotoxin (1 μM) was much greater in diabetic neurons compared with normal animals.These data suggest that under diabetic conditions the activity of N-and L- but not T-type voltage-gated Ca²⁺ channels substantially increased in dorsal horn neurons.     

Immunocytochemical evidence that GABA and neurotensin exist in different neurons in laminae II and III of rat spinal dorsal horn.

Pre-embedding immunocytochemistry with antiserum to neurotensin was combined with post-embedding immunocytochemistry with GABA antiserum, in order to identify neurotensin- and GABA-containing neurons in laminae I-III of rat spinal dorsal horn. The distribution of cell bodies containing these two compounds was similar to that which has been described previously. None of the 88 neurotensin-immunoreactive neurons which were tested showed GABA-like immunoreactivity, which suggests that GABA and neurotensin exist in different cells in this region. Since both compounds are thought to be present in islet cells, it is likely that there are two neurochemically distinct populations of islet cells in lamina II of rat spinal cord.     

Bicuculline-resistant, Cl- dependent GABA response in the rat spinal dorsal horn.

Receptors for gamma-aminobutyric acid (GABA), the main inhibitory neurotransmitter in the mammalian central nervous system (CNS), have been divided into three subtypes. GABA(A) receptor is a ligand-gated chloride channel that is competitively antagonized by bicuculline, whereas GABA(B) receptor regulate Ca2+ or K+ channels through G proteins. Recently, GABA(C) receptor has been identified in mammalian and fish retina. Unlike GABA(A) receptors, the GABA(C) receptor is a bicuculline-resistant chloride channel that is selectively activated by cis-4-aminocrotonic acid (CACA), and antagonized by imidazole-4-acetic acid (I4AA) and to some extent by picrotoxin. We report here that bicuculline-resistant GABA responses mediated by chloride channels are also expressed in substantia gelatinosa (SG) neurons in the dorsal horn, which receive predominantly nociceptive inputs from periphery. The GABA responses are, however, not mimicked by CACA nor affected by I4AA, but abolished by picrotoxin. Moreover, these responses are modulated by benzodiazepines (flunitrazepam) and barbiturates (thiopental), although GABA(C) responses are not affected. Thus, the pharmacological characteristics of the GABA responses observed in SG neurons are distinct from those responses mediated by the known GABA receptors. These differences may reflect the presence of receptor subunits unique to SG neurons.     

High potassium-induced facilitation of glycine release from presynaptic terminals on mechanically dissociated rat spinal dorsal horn neurons in the absence of extracellular calcium

The high potassium-induced potentiation of spontaneous glycine release in extracellular Ca2+-free conditions was studied in mechanically dissociated rat spinal dorsal horn neurons using whole-cell patch-clamp technique. Elevating extracellular K+ concentration reversibly increased the frequency of spontaneous glycinergic inhibitory postsynaptic currents (IPSCs) in the absence of extracellular Ca2+. Blocking voltage-dependent Na+ channels (tetrodotoxin) and Ca2+ channels (nifedipine and omega-grammotoxin-SIA) had no effect on this potassium-induced potentiation of glycine-release. The high potassium-induced increase in IPSC frequency was also observed in the absence of extracellular Na+, although the recovery back to baseline levels of release was prolonged under these conditions. The action of high potassium solution on glycine release was prevented by BAPTA-AM, by depletion of intracellular Ca2+ stores by thapsigargin and by the phospholipase C inhibitor U-73122. The results suggest that the elevated extracellular K+ concentration causes Ca2+ release from internal stores which is independent of extracellular Na+- and Ca2+-influx, and may reveal a novel mechanism by which the potassium-induced depolarization of presynaptic nerve terminals can regulate intracellular Ca2+ concentration and exocytosis.     

Mechanisms of modulation of pregnanolone on glycinergic response in cultured spinal dorsal horn neurons of rat

The glycine receptors and neurosteroids in spinal cord are both implicated in nociceptive signal processing. However, the modulatory effects of neurosteroid pregnanolone (5beta-pregnan-3alpha-ol-20-one) on native glycine receptors remain unclear. In the present study, we examined the effects of pregnanolone and its three isomers on glycine receptors by using whole-cell patch-clamp technique. Our results showed that pregnanolone reversibly inhibited the amplitude of glycine-induced current mediated by native glycine receptors and recombinant alpha1-, alpha2-, alpha3- and alpha1beta-glycine receptors. In cultured spinal dorsal horn neurons of rats, pregnanolone inhibited the glycine-induced current in dose-dependent manner, with an antagonist concentration inducing half-maximal response of 1.0+/-0.3 microM. The inhibitory effect of pregnanolone on glycine-induced current was voltage-independent and pregnanolone shifted the concentration-response curve for glycine-induced current rightward in a parallel manner without altering the maximal value and Hill coefficient. The isomer of pregnanolone, allopregnanolone (5alpha-pregnan-3alpha-ol-20-one) slightly enhanced glycine-induced current, whereas iso-pregnanolone (5beta-pregnan-3beta-ol-20-one) and iso-allopregnanolone (5alpha-pregnan-3beta-ol-20-one) did not affect the glycine-induced current significantly in cultured spinal dorsal horn neurons. Thus, our results suggest that the inhibitory effect of pregnanolone on glycine-induced current is of a competitive type and depends on the stereo structure of pregnanolone. Furthermore, pregnanolone decreased the amplitude and frequency of the glycinergic miniature inhibitory postsynaptic currents. Through modulating the glycinergic inhibitory neurotransmission, pregnanolone may affect the nociceptive sensory processing under physiological and pathological conditions.     

Immunohistochemical evidence that acetylcholine and glycine exist in different populations of GABAergic neurons in lamina III of rat spinal dorsal horn.

Pre-embedding immunohistochemistry with monoclonal antibody to choline acetyltransferase was combined with post-embedding immunohistochemistry with antisera to GABA and glycine in order to study the pattern of coexistence of GABA, glycine and acetylcholine in neurons in lamina III of rat spinal dorsal horn. Of 50 neurons which were choline acetyltransferase immunoreactive, 47 showed GABA-like immunoreactivity and none were immunoreactive with antiserum to glycine, despite the fact that glycine is thought to be present in the majority of GABAergic neurons in lamina III. This suggests that while acetylcholine and glycine can both coexist with GABA in lamina III neurons, they are present in different populations of GABAergic cells. Taken together with recent ultrastructural evidence concerning the synaptic connections of glycinergic and cholinergic structures in the dorsal horn, this suggests that there are functional differences between neurons which contain GABA and glycine and those which contain GABA and acetylcholine.     

Sustained L-type calcium currents in dissociated deep dorsal horn neurons of the rat: characteristics and modulation

Deep dorsal horn neurons present plateau properties involved in non-linear integration of nociceptive inputs, in the windup of the discharge, and in the expression of long-lasting afterdischarges. In vitro experiments using intracellular recordings in a slice preparation of the rat spinal cord have established that they are supported in part by voltage-dependent calcium currents, and positively modulated by metabotropic glutamate receptor activation. In the present study, whole-cell patch-clamp recordings in acutely isolated soma of dorsal horn neurons (n=48) were used to analyse the voltage-dependent calcium currents involved.Deep dorsal horn neurons expressed both inactivating and non-inactivating calcium currents with Ca(2+) or Ba(2+) used as a charge carrier. The non-inactivating component activated at intermediate threshold (-55mV), and was blocked mostly by nifedipine (61+/-6%). Although voltage-dependent facilitation of whole-cell calcium currents could be obtained by prepulses to +100mV, repetitive depolarization at potentials compatible with the plateau (-45mV and -10mV) failed to induce facilitation of calcium currents. No direct modulation of somatic calcium currents by application of (S)-3,5-dihydroxyphenylglycine, a selective group I metabotropic glutamate receptor agonist and 1S,3R-1-amino-1,3-cyclopentanedicarboxylic acid, a group I and II metabotropic glutamate receptor agonist, was found, while application of the metabotropic GABA(B) receptor agonist baclofen induced a significant decrease in calcium currents.Thus, the present voltage-clamp study shows that rat deep dorsal horn neurons express a non-inactivating, nifedipine sensitive, intermediate threshold (-55mV) calcium current which could provide the depolarizing drive to generate plateau potentials near threshold. Our results also indicate that calcium currents are not sensitized following repetitive stimulation, and not modulated by metabotropic glutamate receptor activation. They provide, however, the first evidence for a direct modulation of voltage-gated calcium channels in dorsal horn neurons by GABA(B) receptor activation, which may contribute to the mechanism of baclofen's antinociceptive activity.     

Convergent control of synaptic GABA release from rat dorsal horn neurones by adenosine and GABA autoreceptors

Perforated patch clamp recordings were performed on cultured superficial neonatal rat dorsal horn (DH) spinal cord neurones in order to study the presynaptic modulation of GABA release at unitary synaptic connections. Since ATP can be coreleased with GABA at about two-thirds of GABAergic synapses between DH neurones, and can be rapidly metabolized to adenosine in the extracellular space, we investigated the potential role of A1 adenosine receptors and GABA_B receptors which might function as inhibitory autoreceptors. Adenosine and GABA_B receptor agonists reduced the amplitude of electrically evoked GABAergic inhibitory postsynaptic currents (eIPSCs) as well as the frequency of GABAergic miniature IPSCs, suggesting a presynaptic action of these substances. The actions of adenosine were blocked by the A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). The effects of adenosine and GABA_B agonists were occlusive, indicating a functional convergence of the signalling pathways engaged by A1 and GABA_B receptors. A1 and GABA_B antagonists increased the amplitude of eIPSCs in a supra-additive manner, suggesting a tonic activation of these receptors by ambient adenosine and GABA. Moreover, using trains of electrical stimulations, we were able to unravel a phasic (activity-dependent) activation of presynaptic A1 and GABA_B autoreceptors only in the case of neurones coreleasing ATP and GABA, despite the presence of functional presynaptic A1 and GABA_B receptors on all GABAergic DH neurones. This selective, convergent and activity-dependent inhibition of GABA release by A1 and GABA_B autoreceptors might modulate the integrative properties of postsynaptic DH neurones under physiological conditions and/or during the development of pathological pain states.     

GABA-immunoreactive neurons in the dorsal horn of the rat spinal cord

An antiserum to GABA was used on semithin resin-embedded sections of rat dorsal horn. Immunoreactive neurons were evenly distributed throughout laminae I-III and constituted between 24 and 33% of the total neuronal population within three laminae. Fifty Golgi-stained cells in lamina II were tested with the antiserum. Most of the islet cells examined were immunoreactive, although some small islet cells were not. None of the 14 stalked cells tested was immunoreactive. These results provide further evidence that the stalked and islet cells of lamina II form two distinct functional classes and suggest that the islet cells function as inhibitory interneurons.     

Noradrenergic receptor mRNA expression in adult rat superficial dorsal horn and dorsal root ganglion neurons

Noradrenaline (NAdr) has well documented analgesic actions at the level of the spinal cord. Released from bulbospinal projections onto superficial dorsal horn (SDH) neurons, NAdr modulates the excitability of these neurons through the activation of ₁, ₂ or β adrenoceptors. This study utilised in situ hybridisation to determine the specific expression of adrenoceptors within adult rat lumbar SDH and dorsal root ganglion (DRG) neurons, and reports the presence of _1A, _1B, _2B, β₁ and β₂ adrenoceptor mRNA within SDH neurons, and the presence of _1A, _1B and _2C adrenoceptor mRNA within DRG neurons. The present study provides an insight into the modulation of sensory processing at the level of the spinal cord following adrenoceptor activation.     

Modulation of excitatory amino acid responses in rat dorsal horn neurons by tachykinins.

1. In freshly isolated spinal dorsal horn (DH) neurons (laminae I-III) of the young rat, the effects of tachykinins (substance P, neurokinin A) on inward current induced by excitatory amino acids were studied under whole-cell voltage-clamp conditions. 2. When the cells were clamped to a holding potential of -60 mV, a simultaneous application of N-methyl-D-aspartate (NMDA) (10(-4) M) and substance P (SP) (2 x 10(-9)-10(-7) M) for 10 s reversibly enhanced (by 129.6 +/- 8.2%, mean +/- SE) the peak amplitude of the initial transient component of the NMDA-induced current in approximately 60% of the examined cells and reduced it (to 83.3 +/- 2.7%) in 27% of the cells. In addition, SP produced an increase (by 133.6 +/- 11.7%) or a small decrease (to 85.9 +/- 1.4%) in the steady-state component of the NMDA response. In difference to SP, a simultaneous application of NMDA (10(-4) M) and neurokinin A (NKA) (10(-10)-10(-7) M) reversibly suppressed (to 86.8 +/- 2.1%) the peak amplitude of the NMDA-induced current in 75% of the examined cells. 3. The NMDA-induced currents were modulated by tachykinins not only during the coadministration but up to 20 min after the removal of the peptide. SP potentiated the initial peak NMDA current by 147.9 +/- 8.1% in 78% of examined cells and decreased it (76.3 +/- 5.7%) in 11% of cells. The potentiating effect was concentration-dependent (range: 10(-11)-10(-8) M) and reversible, but it was reduced with repeated applications. In addition, SP increased (by 125.4 +/- 3.6%) or reduced (to 86.0 +/- 1.8%) the steady-state component of the NMDA response. 4. When the single DH neurons were exposed to SP or NKA for 30 s-7 min before the testing of the NMDA responses, tachykinins had two distinct effects on the peak amplitude of the transient component of the NMDA-induced current, consisting of an initial depression (SP: to 64.8 +/- 2.1%; NKA: to 76.3 +/- 4.4%) followed by a potentiation (SP: by 146.6 +/- 6.8%; NKA: by 178.4 +/- 35.2%). The enhancing effect in some cells lasted less than or equal to 1 h. 5. A claimed novel nonselective tachykinin antagonist, spantide II (10(-8) M) coadministered with NMDA (10(-4) M), slightly depressed the peak component of NMDA-induced current. In addition, it effectively blocked the SP-induced potentiation of the responses of DH neurons to NMDA.(ABSTRACT TRUNCATED AT 400 WORDS)     

Changes in calcium signalling in dorsal horn neurons in rats with streptozotocin-induced diabetes

Intracellular calcium signalling was studied in the dorsal horn from neurons of rats with streptozotocin-induced diabetes versus control animals. The cytoplasmic Ca2+ concentration ([Ca2+]i) was measured in Fura-2 acetoxymethyl ester-loaded dorsal horn neurons from acutely isolated spinal cord slices using a fluorescence technique. The recovery of depolarization-induced [Ca2+]i increase was delayed in diabetic neurons compared with normal animals. In normal neurons, [Ca2+]i after the end of KCl depolarization recovered to the basal level monoexponentially with a time constant of 8.0+/-0.5 s (n = 23), while diabetic neurons showed two exponentials in the [Ca2+]i recovery. The time constants of these exponentials were 7.2+/-0.5 and 23.0+/-0.6 s (n = 19), respectively. The amplitude of calcium release from caffeine-sensitive endoplasmic reticulum calcium stores became significantly smaller in diabetic neurons. The amplitudes of [Ca2+]i transients evoked by 30 mM caffeine were 268+/-29 nM (n = 13) and 31+/-9 nM (n = 17) in control and diabetic neurons, respectively. We conclude that streptozotocin-induced diabetes is associated with prominent changes in the mechanisms responsible for [Ca2+]i regulation, which presumably include a slowdown of Ca2+ elimination from the cytoplasm by the endoplasmic reticulum.     

Superficial dorsal horn neurons with double spike activity in the rat

Superficial dorsal horn neurons promote the transfer of nociceptive information from the periphery to supraspinal structures. The membrane and discharge properties of spinal cord neurons can alter the reliability of peripheral signals. In this paper, we analyze the location and response properties of a particular class of dorsal horn neurons that exhibits double spike discharge with a very short interspike interval (2.01+/-0.11 ms). These neurons receive nociceptive C-fiber input and are located in laminae I-II. Double spikes are generated spontaneously or by depolarizing current injection (interval of 2.37+/-0.22). Cells presenting double spike (interval 2.28+/-0.11) increased the firing rate by electrical noxious stimulation, as well as, in the first minutes after carrageenan injection into their receptive field. Carrageenan is a polysaccharide soluble in water and it is used for producing an experimental model of semi-chronic pain. In the present study carrageenan also produces an increase in the interval between double spikes and then, reduced their occurrence after 5-10 min. The results suggest that double spikes are due to intrinsic membrane properties and that their frequency is related to C-fiber nociceptive activity. The present work shows evidence that double spikes in superficial spinal cord neurones are related to the nociceptive stimulation, and they are possibly part of an acute pain-control mechanism.