Effects of N- and L-type calcium channel antagonists on the responses of nociceptive spinal cord neurons to mechanical stimulation of the normal and the inflamed knee joint.

1. The present study addresses the involvement of voltage-dependent calcium channels of the N and L type in the spinal processing of innocuous and noxious input from the knee joint, both under normal conditions and under inflammatory conditions in which spinal cord neurons become hyperexcitable. In 30 anesthetized rats, extracellular recordings were performed from single dorsal horn neurons in segments 1-4 of the lumbar spinal cord. All neurons had receptive fields in the ipsilateral knee joint. In 22 rats, an inflammation was induced in the ipsilateral knee joint by kaolin and carrageenan 4-16 h before the recordings. The antagonist at N-type calcium channels, omega-conotoxin GVIA (omega-CTx GVIA), was administered topically in solution to the dorsal surface of the spinal cord at the appropriate spinal segments in 6 rats with normal joints and in 12 rats with inflamed knee joints. The antagonist at L-type channels, nimodipine, was administered topically in 5 rats with normal joints and in 11 rats with inflamed knee joints. In another five rats with inflamed joints, antagonists at L-type calcium channels (diltiazem and nimodipine) and omega-CTx GVIA were administered ionophoretically with multibarrel electrodes close to the neurons recorded. 2. The topical administration of omega-CTx GVIA to the spinal cord reduced the responses to both innocuous and noxious pressure applied to the knee joint in a sample of 11 neurons with input from the normal joint and in a sample of 16 neurons with input from the inflamed joint (hyperexcitable neurons). The responses were decreased to approximately 65% of the predrug values within administration times of 30 min. A similar reduction of the responses to innocuous and noxious pressure was observed when omega-CTx GVIA was administered ionophoretically to nine hyperexcitable neurons. In neurons with input from the normal or the inflamed knee joint, the administration of omega-CTx GVIA led also to a reduction of the responses to innocuous and noxious pressure applied to the noninflamed ankle joint. 3. The topical administration of nimodipine decreased the responses to innocuous and noxious pressure applied to the knee in a sample of 9 neurons with input from the normal joint and in a sample of 16 neurons with input from the inflamed knee joint (hyperexcitable neurons). Within administration times of 30 min, the responses were reduced to approximately 70% of the predrug values. In hyperexcitable neurons, the responses to innocuous and noxious pressure applied to the knee were also decreased during ionophoretic administration of nimodipine (6 neurons) and diltiazem (9 neurons). When the noninflamed ankle was stimulated, the responses to innocuous pressure were reduced neither in neurons with input from the normal knee nor in neurons with input from the inflamed knee, but the responses of hyperexcitable neurons to noxious pressure onto the ankle were reduced. The ionophoretic administration of the agonist at the L-type calcium channel, S(-)-Bay K 8644, enhanced the responses to mechanical stimulation of the knee joint in all 14 hyperexcitable neurons tested. The effect of S(-)-Bay K 8644 was counteracted by both diltiazem (in 6 of 6 neurons) and nimodipine (in 5 of 5 neurons). 4. These data show that antagonists at both the N- and the L-type voltage-dependent calcium channels influence the spinal processing of input from the knee joint. The data suggest, therefore, that voltage-dependent calcium calcium channels of both the N and the L type are important for the sensory functions of the spinal cord. They are involved in the spinal processing of nonnociceptive as well as nociceptive mechanosensory input from the joint, both under normal and inflammatory conditions. The present results show in particular that N- and L-type channels are likely to be involved in the generation of pain evoked by noxious mechanical stimulation in normal tissue as well as in the mechanical hyperalgesia that is usually pres

Calcitonin gene-related peptide is involved in the spinal processing of mechanosensory input from the rat's knee joint and in the generation and maintenance of hyperexcitability of dorsal horn-neurons during development of acute inflammation.

In an electrophysiological study in anaesthetized rats, the involvement of calcitonin gene-related peptide in the spinal processing of mechanosensory information from the normal and inflamed knee joint was investigated. Calcitonin gene-related peptide(8-37), a specific antagonist at calcitonin gene-related peptide 1 receptors was administered ionophoretically close to nociceptive neurons with input from the knee joint before, during, and after development of acute inflammation in the knee induced by the intra-articular injections of kaolin and carrageenan. Calcitonin gene-related peptide (8-37) selectively antagonized the effects of ionophoretically applied calcitonin gene-related peptide but not those of ionophoretically applied substance P, neurokinin A, and (R,S)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid. Before inflammation, calcitonin gene-related peptide (8-37) reduced the responses to noxious pressure applied to the knee in 22 of 23 neurons; in 14 of 22 neurons, the responses to innocuous pressure were also reduced. In eight neurons calcitonin gene-related peptide (8-37) was administered during induction and in three periods within the first 90 min of inflammation. In these neurons the developing inflammation evoked a significantly smaller increase of the responses to innocuous and noxious pressure applied to the injected knee than in 13 control neurons which were not treated by the antagonist during induction of inflammation. In 16 of 16 neurons, calcitonin gene-related peptide (8-37) reduced the responses to innocuous and noxious pressure once inflammation and hyperexcitability of the spinal cord neurons were established. These data show that calcitonin gene-related peptide is involved in the spinal processing of mechanosensory input from the normal joint. Furthermore, this peptide and its spinal receptors significantly contribute to the generation and expression of inflammation-evoked hyperexcitability of spinal cord neurons during the development of inflammation. Finally, calcitonin gene-related peptide is involved in the maintenance of inflammation-evoked hyperexcitability. By these effects calcitonin gene-related peptide receptors may significantly contribute to the neuronal basis of hyperalgesia and allodynia associated with inflammation.

The role of spinal neurokinin-2 receptors in the processing of nociceptive information from the joint and in the generation and maintenance of inflammation-evoked hyperexcitability of dorsal horn neurons in the rat.

In spinal cord neurons in anesthetized rats, the role on neurokinin A and neurokinin-2 receptors in the processing of nociceptive information from the knee joint was studied. The specific non-peptide antagonist at the neurokinin-2 receptor, SR48968, its inactive R-enantiomer, SR48965, neurokinin A, substance P and (RS)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA), were administered ionophoretically close to neurons with input from the knee joint. SR48968 reduced the effects of exogenous neurokinin A, but not those of exogenous substance P and AMPA, indicating selective blockade of neurokinin-2 receptors. In most neurons with input from the normal knee joint, SR48968 reduced dose-dependently the responses to noxious pressure with applied to the knee, and in approximately 50% of the neurons the responses to innocuous pressure. The administration of SR48968 during the induction of an experimental joint inflammation markedly attenuated the development of inflammation-evoked hyperexcitability. In hyperexcitable neurons with input from the inflamed joint, SR48968 reduced the responses to noxious and innocuous pressure. The relative reduction of the responses was more pronounced than in neurons with input from the normal joint. None of the effects of SR48968 was mimicked by SR48965. These data show that neurokinin-2 receptors are involved in the spinal processing of nociceptive information from the normal joint. Furthermore, neurokinin-2 receptors must be coactivated at an early stage of inflammation, to allow the generation of hyperexcitability. Finally, neurokinin-2 receptors are involved in maintenance of hyperexcitability during inflammation. In summary, spinal neurokinin-2 receptors are important in the generation of pain in the normal and inflamed joint.

Antinociceptive effects of R(-)- and S(+)-flurbiprofen on rat spinal dorsal horn neurons rendered hyperexcitable by an acute knee joint inflammation.

The antinociceptive effects of the S(+)-enantiomer of flurbiprofen (potent inhibitor of cyclooxygenase) and the R(-)-enantiomer (500 times less potent) were investigated in the spinal cord of 20 anesthetized rats. In lumbar segments, 20 wide-dynamic-range dorsal horn neurons with knee joint input were recorded extracellularly. After induction of an acute inflammation in the knee joint by kaolin and carrageenan, the neurons developed hyperexcitability consisting of enhanced responses to stimuli applied to the inflamed knee and the noninflamed ankle and an expansion of receptive fields. Intravenous administration of R(-)-flurbiprofen (1-9 mg/kg) and S(+)-flurbiprofen (0.3-9 mg/kg) at 5.5 to 8.5 h after kaolin, dose dependently reduced the neurons' responses to pressure applied to the inflamed knee (18 of 18 neurons) and the noninflamed ankle (17 of 17 neurons) and paw (8 of 8 neurons). R(-)-flurbiprofen decreased the receptive field size in 8 of 16 neurons, S(+)-flurbiprofen in 10 of 16 neurons. The suppressive effects started 3 to 6 min and reached a maximum 9 to 15 min after i.v. administration. S(+)-flurbiprofen was more potent than the R(-)-enantiomer. When injected directly into the knee joint, S(+)-flurbiprofen (50 and 80 micrograms), but not the R(-)-enantiomer (100 and 180 micrograms) reduced the hyperexcitability in 12 of 12 neurons. These results suggest a central site of antinociceptive action for R(-)-flurbiprofen and S(+)-flurbiprofen and an additional peripheral site for S(+)-flurbiprofen. The doses used in these experiments did not produce any sedative effects in rats subjected to behavioral testing.