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Annals of the New York Academy of Sciences 12/2006; 448(1):385 - 402. · 3.15 Impact Factor
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ABSTRACT: We investigated the neuronal plasticity in the spinal dorsal horn and its relationship with spinal inhibitory networks using an optical-imaging method that detects neuronal excitation. High-intensity single-pulse stimulation of the dorsal root activating both A and C fibers evoked an optical response in the lamina II (the substantia gelatinosa) of the dorsal horn in transverse slices of 12- to 25-day-old rat spinal cords stained with a voltage-sensitive dye, RH-482. The optical response, reflecting the net neuronal excitation along the slice-depth, was depressed by 28% for more than 1 h after a high-frequency conditioning stimulation of A fibers in the dorsal root (3 tetani of 100 Hz for 1 s with an interval of 10 s). The depression was not induced in a perfusion solution containing an NMDA antagonist, DL-2-amino-5-phosphonovaleric acid (AP5; 30 microM). In a solution containing the inhibitory amino acid antagonists bicuculline (1 microM) and strychnine (3 microM), and also in a low Cl(-) solution, the excitation evoked by the single-pulse stimulation was enhanced after the high-frequency stimulation by 31 and 18%, respectively. The enhanced response after conditioning was depotentiated by a low-frequency stimulation of A fibers (0.2-1 Hz for 10 min). Furthermore, once the low-frequency stimulation was applied, the high-frequency conditioning could not potentiate the excitation. Inhibitory transmissions thus regulate the mode of synaptic plasticity in the lamina II most likely at afferent terminals. The high-frequency conditioning elicits a long-term depression (LTD) of synaptic efficacy under a greater activity of inhibitory amino acids, but it results in a long-term potentiation (LTP) when inhibition is reduced. The low-frequency preconditioning inhibits the potentiation induction and maintenance by the high-frequency conditioning. These mechanisms might underlie robust changes of nociception, such as hypersensitivity after injury or inflammation and pain relief after electrical or cutaneous stimulation.
Journal of Neurophysiology 05/2000; 83(4):2412-20. · 3.32 Impact Factor
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ABSTRACT: The neuronal plasticity in the spinal dorsal horn induced after conditioning low-frequency stimulation of afferent A fibers, and its relationship with spinal inhibitory networks, was investigated with an optical-imaging method that detects neuronal excitation. High-intensity single-pulse stimulation of the dorsal root activating both A and C fibers evoked an optical response in the dorsal horn in transverse slices of 12- to 25-day-old rat spinal cords stained with a voltage-sensitive dye, RH-482. The optical response, reflecting the net excitation of neuronal elements along the thickness of each slice, was suppressed after a conditioning low-frequency stimulation (0.2-1 Hz for 10 min) to A fibers in the dorsal root. The degree of suppression was largest in the lamina II of the dorsal horn (48% reduction), where the majority of C fibers terminate, and much less in the deeper dorsal horn (5% reduction in laminae III-IV). The onset of suppression was somewhat slow; after the low-frequency stimulation, the magnitude of excitation gradually decreased, reached the maximum effect 30 min after the conditioning, and remained at the suppressed level for >1 h. Suppression was not observed when the low-frequency stimulation was given during a 20-min perfusion with a solution containing an NMDA-receptor antagonist, DL-2-amino-5-phosphonovaleric acid (30 microM). A brief application of an opioid-receptor antagonist, naloxone (0.5 microM), inhibited the induction, but not the maintenance, of low-frequency stimulus-induced suppression. However, treatments with the GABA(A) receptor antagonist bicuculline (1 microM) and the glycine receptor antagonist strychnine (0.3 microM) did not affect suppression induction and maintenance. In conclusion, conditioning low-frequency stimulation to A fibers interferes with the afferent-induced excitation in the dorsal horn. The low-frequency stimulation-induced suppression is maintained by a reduction of glutamatergic excitatory transmissions in the dorsal horn, not by an enhanced inhibition. Activation of the spinal opioid-mediated system by low-frequency stimulation, but not the inhibitory amino acid-mediated system, is necessary to initiate robust suppression. The long-term depression of afferent synaptic efficacy onto excitatory interneurons likely takes the primary role in the robust suppression of neuronal excitation in the dorsal horn.
Journal of Neurophysiology 11/1999; 82(4):1957-64. · 3.32 Impact Factor
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ABSTRACT: The neuronal circuitry through which prolonged excitation is generated in the spinal dorsal horn was investigated using optical imaging of neuronal excitation in transverse slices of rat spinal cords. It is known that tetanic stimulation (20 Hz for 1 s) of the dorsal root that activates both A and C primary afferent fibres elicits slow intrinsic optical signals (IOS) in the dorsal horn, seen most intensely in the substantia gelatinosa (SG), lamina II, and that IOS expresses in part the slow synaptic response recorded intracellularly in dorsal horn neurons. We here report that the slow IOS within the SG were completely abolished after an incision was made at the border between the SG and the deeper laminae, but not after an incision within the deeper dorsal horn of the laminae III-V. The result demonstrates directly that, in order to generate prolonged excitation in the SG, the neuronal elements in the deeper dorsal horn must be intact. Thus, the afferent information might be received first by the deeper elements and then transmitted to the SG region, and/or collaboration between the SG and deeper elements is necessary to maintain prolonged excitation in the SG.
European Journal of Neuroscience 10/1999; 11(9):3355-8. · 3.63 Impact Factor
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ABSTRACT: Tetanic stimulation of high-threshold primary afferent fibers in the dorsal root was found to elicit intrinsic optical signals (IOSs) in transverse slices of 11- to 20-day-old rat spinal cords. The IOS, lasting for 30 s or longer, was most prominent in the lamina II of the dorsal horn. Treatment with a Na+-K+-2Cl- co-transport blocker, furosemide, abolished the IOS, suggesting that the origin of the IOS is the cellular swelling due to an activity-dependent rise in extracellular K+. Substance P antagonist spantide, glutamate antagonists 2-amino-5-phosphonovaleric acid and 6-cyano-7-nitroquinoxaline-2,3-dione, and the mu-opioid agonist [d-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin suppressed IOSs. Thus, IOSs represent at least in part the slow excitatory response that is known to be generated in dorsal horn neurons after tetanic activation of unmyelinated afferent fibers.
Neuroreport 12/1998; 9(16):3663-7. · 1.66 Impact Factor
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ABSTRACT: Optical imaging with high spatial and temporal resolution of neural activity in rat cortical slices was used to investigate the dynamics of signal transmission through neural connections in the visual cortex. When inhibition due to gamma-aminobutyric acid was slightly suppressed, horizontal propagation of excitation in both the supra- and infragranular layers became prominent. This propagation was not affected by vertical cuts in either the supra- or infragranular layer, which suggests that excitation is at least partially conveyed horizontally by reciprocal vertical connections between neurons in these layers.
Science 12/1994; 266(5187):1057-9. · 31.20 Impact Factor
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ABSTRACT: The ATP-sensitive K+ channel (KATP channel) is a K+ channel inhibited by cytoplasmic ATP. It was originally found in cardiac cells and recently in neuronal cells. Here, we present evidence indicating that the KATP channel also exists in spinal dorsal horn neurons: membrane currents were recorded by whole-cell voltage-clamp in spinal dorsal horn neurons isolated from young rats. The outward current was augmented by KATP channel activators nicorandil and minoxidil and reduced by the blocker glibenclamide. This glibenclamide-induced change in the current was augmented when the intracellular ATP was lowered and the reversal potential was shifted according to the external K+ concentration.
Neuroscience Letters 05/1994; 170(2):208-12. · 2.11 Impact Factor
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ABSTRACT: In acutely isolated spinal dorsal horn neurons of the rat, effects of serotonin (5-hydroxytryptamine, 5-HT) on inward current induced by excitatory amino acids were studied under whole-cell voltage-clamp condition. 5-HT suppressed the response to N-methyl-D-aspartate (NMDA), but not the response to kainate or quisqualate. This inhibitory effect of 5-HT on NMDA response was present at 5-HT concentrations as low as 10(-15) M. Although the 5-HT effect exhibited similar pharmacology to the 5-HT1A-type receptors, it was not mimicked by increasing intracellular concentration of adenosine 3',5'-cyclic monophosphate that is the common second messenger for 5-HT1A receptors in the mammalian central nervous system. Glycine strongly antagonized this inhibitory effect of 5-HT, and 5-HT reduced opening of NMDA-gated single channels recorded from the outside-out membrane patch. These lines of evidence are consistent with a possibility that 5-HT might directly modulate the NMDA receptor-ion channel complex, either by interacting with the regulatory site(s) or by acting on a distinct site.
Brain Research 09/1990; 525(1):84-91. · 2.73 Impact Factor
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ABSTRACT: The effect of antagonists on N-methyl-D-aspartate (NMDA)-induced response was investigated in isolated nucleus tractus solitarii (NTS) neurons freshly isolated from the rat using a conventional pathclamp technique. The NMDA-induced inward current consisted of an initial peak followed by a steady-state component. The competitive antagonists of NMDA receptor, D-2-amino-5-phosphonovalerate (APV), D-2-amino-4-phosphonoheptanoate (APH) and 3-3(2-carboxypiperazine-4-yl)propyl-1-phosphate (CPP), selectively suppressed the initial peak of NMDA-induced current more than the steady-state component at low concentrations. The non-competitive antagonists, MK-801, ketamine, Zn2+ and Mg2+, equally blocked both peak and steady-state components.
Neuroscience Letters 06/1990; 113(2):169-74. · 2.11 Impact Factor
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ABSTRACT: Acutely isolated rat central neurons were recorded by whole-cell voltage-clamp and responses to a class of excitatory amino acid N-methyl-D-aspartate (NMDA) were examined. Rapid application of NMDA evoked inward current consisted of a fast initial peak followed by a sustained component. Glycine potentiated both initial and desensitized states of the NMDA response with identical concentration-dependence. The initial response, but not the sustained component, was abolished when low concentration of NMDA was pre-applied, and glycine could not reverse the desensitization. This evidence suggests that the NMDA receptor desensitization is sensitive to NMDA but not to glycine, and support the hypothesis that glycine initiates the activation of NMDA receptors rather than that glycine prevents desensitization at NMDA receptors in these cells.
Neuroscience Letters 02/1990; 108(1-2):93-8. · 2.11 Impact Factor
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ABSTRACT: The cholinoceptive properties of dorsal horn neurons (lamina III-V) were investigated by means of intracellular recordings from the rat isolated spinal cord slice preparation. In half of the neurons investigated, acetylcholine (ACh) evoked a dose-dependent slow depolarization and increase in excitability; hyperpolarization was observed in 10% of neurons. Acetyl-beta-methylcholine (MCh) similarly depolarized 39% and hyperpolarized 25% of neurons tested; depolarization was also observed following bethanechol. Responses to the muscarinic agonists were abolished by atropine (10(-5) M). Nicotine depolarized 84% of tested neurons; dihydro-beta-erythroidine (5 x 10(-5) M) and (+)-tubocurarine (10(-6) M) antagonized this depolarization. ACh-, MCh- and nicotine-induced depolarizations, associated with changes in input resistance, were maintained in the presence of tetrodotoxin (10(-6) M). Substance P, as well as repetitive electrical stimulation of the dorsal root, also evoked depolarization in ACh-sensitive neurons. Atropine, but not (+)-tubocurarine, diminished responses to both substance P and dorsal root stimulation. These results indicate that dorsal horn neurons are ACh-sensitive and possess both muscarinic and nicotinic receptors. In addition, the parallel sensitivity of neurons to muscarinic agonists, substance P and dorsal root stimulation, as well as the parallel antagonistic effect of atropine, are supportive of a common ionic mechanism underlying the activation of muscarinic and substance P receptors.
Brain Research 11/1989; 500(1-2):12-20. · 2.73 Impact Factor
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ABSTRACT: The responses to excitatory and inhibitory amino acids and peptides were investigated in isolated rat spinal dorsal horn neurons (laminae I-V) of young rats using the whole-cell voltage-clamp technique. The treatment of spinal slices with low concentrations of enzymes and mechanical dissociation yielded isolated neurons that were sensitive to excitatory amino acids (glutamate, kainate, quisqualate and N-methyl-D-aspartate (NMDA), inhibitory amino acids (gamma-aminobutyric acid (GABA), glycine) and peptides (substance P, calcitonin gene-related peptide (CGRP). The responses of dorsal horn neurons to NMDA were potentiated by glycine and CGRP, whereas GABAA responses were enhanced by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA). Our observations indicate that there is reasonable agreement between many of the responses of isolated neurons and those studied in in vivo and in vitro slice and culture preparations.
Neuroscience Letters 09/1989; 103(1):56-63. · 2.11 Impact Factor
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ABSTRACT: 1. The membrane actions of substance P (SP) and a related tachykinin, neurokinin A (NKA), have been investigated by means of a single-electrode, voltage-clamp technique in the immature rat dorsal horn neurons using an in vitro spinal cord slice preparation. 2. When the membrane potential was held at the resting level of between -75 and -55 mV, bath application of SP or NKA (10(-7) to 10(-5) M, for 1-3 min) induced an inward shift in the holding current lasting several minutes. The magnitude of this effect varied between 10 and 400 pA depending on the concentration of the peptides and the holding potential. 3. When a dorsal horn neuron was held at the resting level and subjected to 1-s depolarizing commands to membrane potentials between -60 and -35 mV, slow inward relaxations and inward tail currents, the latter on repolarization to the holding potential, were recorded. During the tachykinin-induced inward shift in the holding current, the inward relaxation and the tail current were augmented in a dose-related manner. 4. The SP-induced augmentation of the slow inward relaxation and the inward tail current is likely to be due to the enhancement of the activation of the Ca2+ current, because the effect was present, and even augmented in a zero-Ca2+, Ba2+-containing solution, it was reduced or completely abolished by zero-Ca2+, Co2+-, or Mg2+-containing solutions and is largely independent of the changes in external Na+, K+, or Cl- ions. Moreover, in the presence of the K+-channel blocker, tetraethylammonium (TEA), the effect is increased. 5. Depolarizing voltage commands to potentials positive to -35 mV evoked a large, outward K+ current response in the dorsal horn neurons, which was in part Ca2+-sensitive. The outward current response was augmented by SP. The SP effect persists, although being reduced in a zero-Ca2+, Ba2+- or Co2+-containing solutions. 6. In a zero-Ca2+ solution containing Co2+ and TEA, the augmentation of the Ca2+ current and the outward K+ current by SP was abolished. However, the SP-induced increase in a Ca2+-sensitive, voltage-insensitive conductance remained, although being reduced, and the response showed a reversal at about -28 mV. This current may be a result of a tachykinin-activated nonspecific increase in cationic permeability of the membrane of dorsal horn neurons, because the current is reduced by more than one-half when Na+ or Ca2+ is removed from the bathing medium.(ABSTRACT TRUNCATED AT 400 WORDS)
Journal of Neurophysiology 05/1989; 61(4):854-65. · 3.32 Impact Factor
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ABSTRACT: The actions of calcitonin gene-related peptide (CGRP) were examined on Ca2+-dependent action potentials and voltage-dependent Ca2+ currents in rat dorsal root ganglion (DRG) neurons in vitro. In addition, we tested the effect of CGRP on excitatory synaptic transmission in the rat spinal dorsal horn. CGRP produced a reversible increase in the amplitude and the duration of the Ca2+ spike of DRG neurons and directly increased the voltage-dependent Ca2+ current by enhancing both the transient and the sustained components of the current. The increase in the Ca2+ current is likely to be responsible for the increase in the Ca2+ spike and facilitation of excitatory synaptic transmission.
Neuroscience Letters 08/1988; 89(3):305-12. · 2.11 Impact Factor
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ABSTRACT: The membrane actions of calcitonin gene-related peptide (CGRP) and the effect on the Ca-dependent action potential of dorsal horn neurons have been investigated by means of an intracellular recording technique in the immature rat in vitro spinal cord slice-dorsal root ganglion preparation. Bath application of CGRP (10(-8)-10(-6) M for 1-10 min) produced a slow reversible depolarization in about one-third of the cells examined. Biphasic membrane response consisting of an initial hyperpolarization followed by a late prolonged depolarization was seen in a smaller proportion of tested cells. Both membrane responses were present, and even enhanced, when synaptic transmission and Na spikes were blocked by perfusing the slice with a TTX-containing Krebs solution. The CGRP-induced membrane changes were also present in media containing TTX and TEA. The CGRP-evoked depolarization was associated with an increase in the input resistance, and enhanced excitability in a majority of neurons tested. In addition, CGRP modified the duration of Ca-dependent action potentials of dorsal horn neurons, the most consistent change being a prolonged increase in the spike duration. Our results are consistent with a neurotransmitter or neuromodulator role for CGRP in the rat spinal dorsal horn.
Brain Research 03/1988; 441(1-2):357-61. · 2.73 Impact Factor
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ABSTRACT: The membrane actions of calcitonin gene-related peptide (CGRP) and the effect of CGRP on the Ca-dependent action potential of rat dorsal root ganglion (DRG) neurons have been studied by means of an intracellular recording technique in isolated DRG of 2-3-week-old rats in vitro. Bath application of CGRP (10(-8)-10(-6) M for 1-5 min) elicited a slow reversible hyperpolarization and this hyperpolarizing effect was still observed in the medium containing TTX and TEA. However, about half of the large cells, classified by duration of action potential, were depolarized by CGRP. These membrane effects of CGRP were associated with an increase in membrane input resistance (about 20%). In addition, CGRP increased the duration of Ca-dependent action potentials. Our results are consistent with the role of CGRP as an excitatory neurotransmitter or neuromodulator in DRG-spinal cord.
Physiologia Bohemoslovaca 02/1988; 37(3):259-65.
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ABSTRACT: Rat spinal dorsal horn neurons in slice preparations perfused with Ringer solution containing 0.5-1 microM TTX and/or 10-20 mM tetraethylammonium at 29 degrees C, were studied by using a single microelectrode voltage-clamp technique. Slow persistent inward currents were recorded during depolarizing voltage commands to membrane potentials positive to about -40 mV. The inward current was depressed by removing external Ca, or by adding 0.1-0.2 mM Cd, 5 mM Co or 0.1 mM verapamil, and was increased by adding Ba or Bay-K 8644. Substance P (SP) augmented a persistent slow inward Ca-sensitive current in a dose-dependent manner. It is suggested that this effect may be instrumental in generating the SP-evoked slow depolarization, increase in membrane excitability, and the 'bursting' behavior in the immature rat dorsal horn neurons. In addition, in some neurons SP reduced the M-like current, which effect may contribute to, but not explain, generation of the SP-induced slow depolarization.
Brain Research 03/1986; 365(2):369-76. · 2.73 Impact Factor
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Annals of the New York Academy of Sciences 02/1985; 448:385-402. · 3.15 Impact Factor
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ABSTRACT: The membrane actions of substance P (SP) and the effects on the Ca-dependent action potential of dorsal horn neurones have been investigated by means of intracellular recording techniques in the immature rat in vitro spinal cord slice preparation. Bath application of SP (2 X 10(-6) to 1 X 10(-5) M) induced a biphasic membrane response consisting of an initial hyperpolarization followed by a depolarization in about one-third of the cells examined. Initial hyperpolarization was not observed when synaptic activity was blocked by perfusing the slice with a tetrodotoxin-containing or low Ca, high Mg Ringer solution. This result is consistent with a presynaptic action of SP mediated through excitation of inhibitory interneurones. This interpretation was supported by recording of repetitive spontaneous inhibitory post-synaptic potential (i.p.s.p.)-like hyperpolarizing potentials during the initial hyperpolarization. When Co ions were used to block voltage-dependent Ca conductance and possible indirect presynaptic actions, SP induced only a small depolarization of membrane potential. It seems, therefore, that Ca conductance may have contributed to the depolarizing phase of the SP response, either through its mediation of synaptic transmission or through direct effects as a charge carrier for inward current. When tetrodotoxin was used, the SP-induced increase in neuronal input resistance was not modified, although depolarization was slightly diminished. In contrast, in medium containing tetrodotoxin and tetraethylammonium, the SP-depolarizing response was enhanced and accompanied by a small decrease in input resistance and firing of Ca spikes. These results suggest that SP-induced depolarization might be a consequence of a reduction in a voltage-dependent K conductance allowing Na and/or Ca conductances to dominate. SP modified the duration of Ca-dependent action potentials of dorsal horn neurones, the most consistent change being an initial dose-dependent and reversible decrease in the spike duration. The decrease in Ca spike duration was associated with a small reduction in the rate of rise and peak amplitude, and a significant parallel increase in dV/dt of the falling phase of the Ca spike. Our data indicate that the initial decrease in Ca spike duration was not due to the depolarizing action of SP, although shunting of the membrane resistance, either through presynaptic or post-synaptic mechanisms, has not been ruled out. Alternatively, these data are consistent with the possibility that SP shortens the duration of the Ca spike by decreasing a voltage-sensitive inward Ca current and/or augmenting an outward K current.(ABSTRACT TRUNCATED AT 400 WORDS)
The Journal of Physiology 02/1984; 346:203-17. · 4.72 Impact Factor
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ABSTRACT: 1. The electrophysiological properties of dorsal horn neurones have been investigated in the immature rat in vitro spinal cord slice preparation. 2. Intracellular recordings from dorsal horn neurones show that direct or orthodromic stimulation generates action potentials followed by a brief after-hyperpolarization. Synaptic potentials were elicited by the activation of primary afferent fibres in the dorsal root. 3. Input resistance for dorsal horn neurones ranged from 48 to 267 M omega, and the membrane time constant was in the range of 4-19 ms. 4. In response to strong depolarizing currents dorsal horn neurones perfused with TTX and TEA frequently exhibit a slow regenerative depolarizing potential followed by a slow after-hyperpolarization. The depolarizing potential probably results from an influx of Ca. It is blocked by low concentration Ca, Co or Mn, and enhanced by high levels of extracellular Ca. 5. There is, in addition, a low-threshold Ca-dependent response which is activated at membrane potentials more negative than -65 mV and has a maximum rate of rise at the polarization level of about -80 mV. 6. The addition of Ba or TEA to the perfusing medium provided support for the Ca-dependence of the low- and high-threshold responses, and the lack of fast inactivation of the high-threshold Ca potential.
The Journal of Physiology 02/1983; 334:141-53. · 4.72 Impact Factor
