Actions of endomorphins on synaptic transmission of Adelta-fibers in spinal cord dorsal horn neurons.

The effects of endogenous mu-opioid ligands, endomorphins, on Adelta-afferent-evoked excitatory postsynaptic currents (EPSCs) were studied in substantia gelatinosa neurons in spinal cord slices. Under voltage-clamp conditions, endomorphins blocked the evoked EPSCs in a dose-dependent manner. To determine if the block resulted from changes in transmitter release from glutamatergic synaptic terminals, the opioid actions on miniature excitatory postsynaptic currents (mEPSCs) were examined. Endomorphins (1 microM) reduced the frequency but not the amplitude of mEPSCs, suggesting that endomorphins directly act on presynaptic terminals. The effects of endomorphins on the unitary (quantal) properties of the evoked EPSCs were also studied. Endomorphins reduced unitary content without significantly changing unitary amplitude. These results suggest that in addition to presynaptic actions on interneurons, endomorphins also inhibit evoked EPSCs by reducing transmitter release from Adelta-afferent terminals.

Nitric oxide release from substantia gelatinosa of the rat spinal cord in vitro.

To study characteristics of nitric oxide (NO) release from substantia gelatinosa (SG) in the spinal cord, we measured NO concentration in transverse spinal cord slices of rats using electrochemical NO probes. Electrical stimulation of the dorsomedial white matter adjacent to SG elicited transient current changes in NO probes placed on SG and the amplitude corresponded to a NO concentration of 200-300 pM. This NO release was not affected by the application of antagonists of glutamate or substance P receptors. The NO release in the rats, which were neonatally treated with capsaicin for denervating C-fibers, was significantly smaller than that in control rats. These data suggest that NO is mainly derived from the unmyelinated afferent nerves in the SG of the spinal cord.

Cerebrospinal fluid nitric oxide metabolites in painful diseases.

To elucidate the involvement of NO in pain transmission in humans, we measured NO metabolites (nitrite/nitrate) in the CSF of patients with painful diseases using an NO analyzer based on the Griess method. The nitrite/nitrate levels in patients with degenerative lumbar disease (DLD), but not those with fracture or appendicitis, were significantly higher than those in an age-matched control group. The duration of pain in the DLD group was much longer than that in the fracture or appendicitis group. The nitrite/nitrate levels in the middle-aged and elderly DLD patients depended on the duration of pain. These data probably suggest that the duration of pain is critical for the elevation in nitrite/nitrate levels.

A novel slow excitatory postsynaptic current in substantia gelatinosa neurons of the rat spinal cord in vitro.

Whole-cell patch-clamp recordings were made from neurons in the substantia gelatinosa of adult rat spinal cord slices with attached dorsal root to study a slow synaptic current evoked by focal or dorsal root stimulation. Repetitive focal stimulation with a monopolar electrode positioned within substantia gelatinosa elicited a slow excitatory postsynaptic current preceded by a fast excitatory postsynaptic current in 73 of 83 neurons. A similar slow excitatory postsynaptic current was also elicited by stimulation of A delta afferent fibres. The amplitude of slow excitatory postsynaptic currents was unchanged when the recording electrode contained guanosine-5'-O-(2-thiodiphosphate). The slow excitatory postsynaptic current and current evoked by aspartate revealed similar reversal potentials and showed a marked outward rectification at holding potentials more negative than -30 mV, while the glutamate-induced current exhibited a relatively linear voltage relationship. In addition, the slow excitatory postsynaptic currents were reversibly occluded during the aspartate-induced current but were not occluded during the glutamate-induced current. The slow excitatory postsynaptic currents evoked by focal stimulation were depressed but not abolished by 6-cyano-7-nitroquinoxaline-2,3-dione (10 microM) or by 6-cyano-7-nitroquinoxaline-2,3-dione together with DL-2-amino-5-phosphonopentanoic acid (100 microM). Similarly, the aspartate- and glutamate-induced currents were also resistant to these antagonists. These observations suggest that a transmitter released from interneurons or descending fibres which are activated in part by A delta afferents, mediates a slow excitatory postsynaptic currents in substantia gelatinosa neurons and that an excitatory amino acid is implicated in the generation of the slow excitatory postsynaptic current, although the receptor appears to differ from the known ligand-gated channels. C afferents are unlikely to contribute to the slow excitatory postsynaptic current. This slow synaptic response may participate in the pain pathway and play an important role in the processing of nociceptive information in the spinal dorsal horn.