Z.-Q. Zhao

Ectopic purinergic sensitivity develops at sites of chronic nerve constriction injury in rat.

The possibility that ectopic purinergic sensitivity develops following peripheral nerve injury was investigated in chronic constriction injury (CCI). Spontaneous firing of A-fibers originated from the injury site or from sensory endings of afferents in the contralateral sciatic nerve. ATP injected intravenously excited most of the injured fibers whereas none of the contralateral afferents responded to ATP. The ATP-induced effect was blocked by the P2 receptor antagonist reactive blue 2, but not the P1 receptor antagonist aminophylline. Neither the alpha-adrenoreceptor antagonist phentolamine nor the cyclooxygenase inhibitor indomethacin attenuated the ATP-evoked effect. We conclude that a novel ectopic purinergic sensitivity mediated by P2 receptors develops at sites of the CCI of nerves in the rat, which may contribute to neuropathic pain.

Buthus martensi Karsch agonist of skeletal-muscle RyR-1, a scorpion active polypeptide: antinociceptive effect on rat peripheral nervous system and spinal cord, and inhibition of voltage-gated Na+ currents in dorsal root ganglion neurons

The antinociceptive effect and potential antinociceptive mechanism of Buthus martensi Karsch agonist of skeletal-muscle RyR-1 (BmK AS-1), a scorpion venom derived neurotoxic polypeptide, have been investigated in rats. The results show that: (a) the withdrawal latency to rat plantar radiant heat was increased significantly by 100 and 150% after intrathecal injection of 0.6 and 1.2 microg doses; (b) C components of rat nociceptive flexion reflex were reduced to 72, 50 and 29% after intraplantar injection of 5, 10 and 20 microg doses; (c) both central (spinal cord) and peripheral antinociceptive effects of BmK AS-1 could not be reversed by naloxone; (d) tetrodotoxin-resistant (TTX-R) Na(+) current was depressed to 83.87+/-1.64, 64.73+/-5.43 and 15.85+/-17.63%, and tetrodotoxin-sensitive (TTX-S) Na(+) current was depressed to about 81.27+/-2.5, 49.08+/-8.09 and 9.03+/-12.34% with 0.2, 1.0 and 10 microg/ml BmK AS-1 measured using patch clamp recording in rat small dorsal root ganglion (DRG) neurons, respectively. The results indicate that BmK AS-1 may be a new component with potent antinociceptive activity mediated by modulating TTX-S and TTX-R Na(+) channels.

Endomorphin-1 reduces carrageenan-induced Fos expression in the rat spinal dorsal horn

Intraplantar injection of carrageenan induced significant Fos expression in the superficial and deep spinal dorsal horn at the L(4)-L(5)segments and extensive peripheral edema of the ipsilateral foot in rats. Intraplantar injection of endomorphin-1, endogenous ligand for mu opioid receptor, in the same region produced dose-dependent reduction of carrageenan-induced Fos expression and peripheral edema, which were completely blocked by co-administration of intraplantar injection of naloxone (20 microgram). The systemic injection of the highest dose of endomorphin-1 (50 microgram) had no significant reductory effect on Fos expression and peripheral edema. These results further provided a strong evidence for involvement of mu opioid receptor in peripheral analgesia, particularly in inflammation pain.

Orphanin FQ/nociceptin modulates glutamate- and kainic acid-induced currents in acutely isolated rat spinal dorsal horn neurons

The heptadecapeptide orphanin FQ (OFQ), also known as nociceptin (NOC), is a newly discovered endogenous ligand for the opioid-like G-protein coupled receptor, ORL1. In the present study, the effects of OFQ/NOC on glutamate (Glu), kainic acid (KA) and quisqualic acid (QA) induced currents were examined in isolated rat spinal dorsal horn neurons of young rats using whole-cell patch-clamp techniques. Glu, KA and QA elicited rapid inward currents in 90%, 69%, 83% of tested neurons. OFQ/NOC(0.03 approximately 300 nM) failed to induce any changes of membrane currents, but modulated Glu-, KA- and QA-elicited currents. OFQ/NOC inhibited and potentiated Glu-induced currents in 40.6% and 27.3% of examined cells (n=106) respectively. In about one third examined neurons, OFQ/NOC had no detectable effects on Glu responses. OFQ/NOC also inhibited and enhanced KA- and QA-induced currents (inhibition: KA, 67.1%, n=76; QA, 50%, n=36. Potentiation: KA, 23.7%, n=76; QA, 16.7%, n=36). In about 10% of tested cells, Glu-induced currents were potentiated after the application of OFQ/NOC, and lasted for 20 approximately 30 min. The inhibitory effects of OFQ/NOC on KA and QA responses were naloxone-insensitive. The C-terminal fragment OFQ(8-17) presented same effects on EAA-induced responses. Taken together, OFQ/NOC primarily inhibited Glu-, KA- and QA-induced currents in isolated rat spinal dorsal horn neurons via non-opioid mechanism, which might contribute to nociceptive transmission in the spinal level.

Selective blockade by yohimbine of descending spinal inhibition from lateral reticular nucleus but not from locus coeruleus in rats

The present study was undertaken to compare the effects of the alpha 2-adrenoceptor antagonist yohimbine on inhibition of C-fiber-evoked responses of dorsal horn neurons produced by electrical stimulation of the lateral reticular nucleus (LRN) and the Locus coeruleus (LC) in the rat. In the majority of neurons, C-fiber-evoked responses were significantly inhibited by 53.84 +/- 5.02% and 57.63 +/- 5.79% of control by LRN and LC stimulation, respectively, whereas in less than half of the neurons, A-fiber-evoked responses were reduced by 20.99 +/- 6.06% and 21.78 +/- 4.48% of control, respectively. After systemic or spinal administration of yohimbine, LC-induced inhibition of C-fiber-evoked responses was not affected. In contrast, LRN-induced inhibition was markedly attenuated by yohimbine. The results suggest that alpha 2-adrenoceptors may be involved in mediation of inhibition of spinal nociception induced by stimulation of LRN but not by LC.

Interaction between substance P and excitatory amino acid receptors in modulation of nociceptive responses of cat spinal dorsal horn neurons

Co-effects of microelectrophoretic application of 2-amino-5-phosphonovalerate (APV), ketamine, 6,7-dinitroqinoxaline-2,3-dione (DNQX), kynurenate (Kyn) and spantide on 34 spinal dorsal horn neurons were studied. Co-application of spantide and APV or DNQX produced a synergetic inhibition of responses by tibial stimulation in 10/23 and 3/8 neurons, respectively, and also in 3/8 neurons by administration of DL-sodium homocysteate. Spantide enhanced APV- or ketamine-induced inhibition of C responses of 8/19 neurons to sural stimulation, DNQX induced that of 2/4 neurons to gastrocnemius-soleous stimulation (GS), and Kyn induced that to both sural (8/19) and GS (2/4). The results suggest an interaction of SP and NMDA, non-NMDA receptors in processing spinal nociception.

Release and depletion of substance P by capsaicin in substantia gelatinosa studied with the antibody microprobe technique and immunohistochemistry

Using an antibody microprobe technique, we have detected substance P release from the region of the substantia gelatinosa of the cat during the first, but not the second, 30 min of topical application of capsaicin (1-3%) to the tibial nerve. Immunohistochemical analysis also showed that substance P-like immunoreactivity was markedly reduced in the superficial layer of the dorsal horn 30 min after application of capsaicin. These results indicate that substance P is released and then depleted from primary afferent central terminals following acute application of capsaicin to the peripheral sensory nerve.

Inhibition of nociceptive withdrawal reflex by microinjection of interleukin 2 into rat locus coeruleus

This study was to examine the effects of microinjection of human recombinant interleukin 2 (IL-2) into locus coeruleus (LC) on spinal nociception. Following application of IL-2 (0.1 microl, 10 pM) into LC, the percentage of inhibition of nociceptive C responses of reflex at 3, 9, 15, 21 and 27 min after injection were 88.2 +/-9.4%, 84.0 +/- 11.8%, 89.7 +/- 10.5%, 57.1 +/- 8.7% and 26.3 +/- 12.2%, respectively. Also, the expression of Fos protein in superficial dorsal horn was reduced by 73.01 +/- 13.58% of control (P<0.0001). Naloxone (10 microg, i.p.) completely blocked the IL-2-induced inhibition of C responses. The results clearly show that IL-2 receptors present in LC mediate descending inhibition of the spinal nociception, which may couple with the activation of opioid receptors on LC neurons.

Neomycin blocks capsaicin-evoked responses in rat dorsal root ganglion neurons

Small dorsal root ganglion (DRG) neurons are characterized by sensitivity to capsaicin. In acutely isolated rat DRG neurons, the effect of neomycin, one of the aminoglycoside antibiotics, on capsaicin-evoked current and voltage responses was examined using whole-cell patch-clamp recording. We showed for the first time that neomycin dose-dependently inhibited capsaicin-evoked currents with half-maximal inhibitory concentration at 130.60 microM (n=70). Under current-clamp condition, depolarization and firing rate evoked by capsaicin became weakened when neomycin was perfused to the neurons (n=10). Neomycin had no significant effect on the resting potentials of DRG neurons. These results suggest that neomycin could inhibit capsaicin-sensitive responses in small DRG neurons, which may contribute to neomycin-induced peripheral analgesia.

Neomycin blocks substance P-induced calcium entry in cultured rat spinal cord neurons

Substance P (SP) plays an important role in sensitization of spinal cord neurons. In this study, we investigated SP-induced calcium activities in cultured rat spinal cord neurons with confocal laser scanning microscopy. Our results showed that SP increased [Ca(2+)](i) by calcium entry rather than release from intracellular calcium stores. Neomycin (100 microM), an antagonist of phosphatidylinositol 4,5-bisphosphate (PIP(2)), blocked SP-induced calcium entry. Ca(2+)-free medium induced capacitative entry, which was significantly potentiated by SP. As activation of SP receptor (NK-1) leads to production of inositol 1,4,5-trisphosphate (IP(3)) by PIP(2) turnover, the results indicates that SP-induced calcium entry and SP-potentiated capacitative calcium entry might be mediated or regulated by IP(3)/diacylglycerol (DG) pathway.