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beta-Arrestin-1 (betaArr1) plays a major role in the desensitization and internalization of G protein-coupled receptors. We previously localized betaArr1 in the sensory neurons of rat lumbar 4 and 5 dorsal root ganglia (DRG) and reported the predominant presence of betaArr1 in the small-diameter DRG neurons that are often implicated with nociception. Because of betaArr1's crucial role in regulating the initiation of cellular signaling, in the current study we evaluated the distribution of betaArr1 in the peripheral sensory terminals where various receptors are present. Western blotting confirmed the presence of betaArr1 immunoreactivity in the rat skin. Sciatic nerve ligation demonstrated that betaArr1 is transported peripherally from the DRG, and immunohistochemistry showed betaArr1 immunoreactivity in the glabrous skin of the rat hindpaw. In the glabrous skin, strong betaArr1 immunoreactivity was detected in nerve fibers in the dermal nerve plexus and dermal papillae. Fine varicose immunoreactive fibers were found in the epidermis. In addition, betaArr1 was observed in specialized sensory receptors such as Meissner corpuscles. Our observations thus indicate that betaArr1 may be involved in modulation of specific tactile stimulation from the skin in addition to nociception.

The purposes of this study were to examine responses of superficial (depth <300 microm) and deeper thoracic spinal neurons to chemical stimulation of cardiac afferents and effects of descending influences on these neurons. Extracellular potentials of single T(3)-T(4) neurons were recorded in pentobarbital anesthetized, paralyzed and ventilated male rats. A catheter was placed in the pericardial sac to administer 0.2 ml of a mixture of algogenic chemicals that contained adenosine (10(-3) M), bradykinin, histamine, serotonin, prostaglandin E(2) (10(-5) M). Fifteen of 55 (27%) superficial neurons responsive to intrapericardial chemicals were compared to 80/169 (47%) deeper neurons. All 15 superficial neurons that responded to cardiac afferents were excited (E), whereas 66 deeper neurons were excited, ten were inhibited and four showed excitation-inhibition. Spontaneous activity of superficial neurons with short-lasting excitatory responses was significantly lower than that of deeper neurons (P<0.05). Somatic receptive fields on chest, axilla, arm and upper back areas were found for 77/95 (81%) neurons that responded to intrapericardial chemicals. The proportion of somatic field properties and their sizes in superficial neurons were similar to deeper neurons. After cervical spinal transection, both spontaneous activity and responses to chemical stimulation of cardiac afferents significantly increased in six out of six neurons excited by intrapericardial injections. Results showed that chemical stimulation of cardiac afferents excited superficial T(3)-T(4) spinal neurons, whereas deeper neurons exhibited multiple patterns of responses. Some characteristics of subgroups of superficial neurons were quantitatively different from deeper neurons. Thoracic spinal neurons processing cardiac nociceptive information were under tonic descending inhibition.

Supporting glial cells of the peripheral nervous system include satellite cells of dorsal root ganglia and Schwann cells of peripheral nerves. In the central nervous system, glial cells contain enzymes related to the tricarboxylic acid and glutamine cycles: pyruvate carboxylase, glutamate dehydrogenase, and glutamine synthetase. The present study used immunohistochemistry in the rat peripheral nervous system to determine the cellular distribution of these enzymes along with glutamine. In dorsal root ganglia and peripheral nerves, glutamine and glutamine related enzymes were enriched in satellite and Schwann cells. In the dorsal root ganglia, immunoreactive satellite cells surrounded neurons of all sizes. In peripheral nerve, immunoreactive Schwann cells were most easily observed surrounding large diameter, myelinated axons. These Schwann cells contained immunoreactivity in their cell bodies, nodes of Ranvier, and the rim of cytoplasm outside the myelin sheath. Myelin sheaths were non-immunoreactive. The peripheral glial tricarboxylic and glutamine cycles may be used to produce glutamine for neuronal cell uptake and conversion to glutamate for synaptic transmission. Alternatively, these cycles may function in peripheral glia similar to central nervous system astrocytes for supporting the energy demands of neurons.

The amino acid taurine serves many functions in the nervous system serving as inhibitory neurotransmitter/neuromodulator, neurotrophin, antioxidant, and osmolyte. Taurine levels are increased following brain injury and glucocorticoid administration. Thus, the purpose of this study was to examine spinal taurine concentrations following spinal cord injury (SCI) and methylprednisolone (MP) treatment of SCI. A total of 44 adult male Sprague-Dawley rats were divided into control and lesion groups. Control rats received a T6 vertebral laminectomy while lesioned rats received a laminectomy followed by complete spinal transection. Half of the animals in each group received MP intravenously following sham-operation or SCI. Rats survived for 7 or 30 days and concentrations of taurine in spinal gray and white matter, in spinal segments both near and distant from the injury epicenter, were resolved by HPLC analysis. Taurine levels were increased 7 and 30 days following transection in spinal segments immediately adjacent to the lesion and were further elevated by MP treatment. No increases were seen in far rostral/caudal segments, and MP treatment alone had no effect on spinal taurine levels. These findings demonstrate that spinal injury results in increased taurine concentrations in spinal segments undergoing the greatest degree of cellular reactivity and tissue reorganization and that MP therapy potentiates these increases. These findings are significant in that they further characterize the effects of acute MP therapy in spinal tissue. Since taurine is thought to be involved in neuroprotection and/or regeneration following injury, the potentiation of taurine levels by MP treatment may relate to its therapeutic properties.

Effects of electrically stimulating the left stellate ganglion to activate cardiopulmonary sympathetic afferent (CPSA) fibers were determined on C1-C3 dorsal horn neurons in anaesthetized rats. Fifty-two of 53 dorsal horn neurons affected by CPSA stimulation were excited and one neuron was inhibited. In 6 experiments, dorsal columns and ventrolateral funiculi were sequentially lesioned to determine neuronal pathways involved in CPSA activation of C1-C3 neurons. In 6 additional experiments, spinal transection at rostral C1 was used to determine the contribution of supraspinal relays. We concluded that CPSA input to C1-C3 segments travelled bilaterally in ventrolateral pathways, and that supraspinal relays were not required for CPSA excitation of C1-C3 neurons. These results suggest that neurons in C1-C3 segments might play an important role in processing visceral spinal afferent information.

Retrograde transport of the fluorescent tracer Fluorogold was used in combination with immunohistochemical staining for the enzyme glutaminase to identify putative glutamatergic neurons belonging to the rat spinomesencephalic tract. Glutaminase-like staining in spinal projection neurons suggests that the relay of nociceptive information from the spinal cord to midbrain may involve the excitatory amino acid glutamate.

The spinal cord contains mu, delta and kappa opioid receptors which mediate the antinociceptive effects of opioid agonists administered onto the spinal cord. In this study, we characterized the binding sites for highly-selective mu, delta and kappa opioid radioligands and quantified the distribution of opioid binding sites in rat lumbosacral spinal cord using autoradiography. In sections of rat brain mounted on glass slides, the mu ligand, [3H]sufentanil, bound with high affinity with an apparent Kd of 0.46 nM. The delta ligand, [3H]DPDPE [( D-Pen2.5]-enkephalin), bound with a Kd of 4.31 nM, and the kappa-ligand, [3H]U69593, bound with a Kd of 2.27 nM. Three regions of the spinal gray were targeted for quantification of binding sites by autoradiography. The data indicate that when considered as a percentage of the total opioid binding capacity within a region, the contribution of mu sites in laminae I-II was about 90%, with delta and kappa sites 7% and 3%, respectively. In lamina V, the mu sites comprised about 70% of the total opioid sites, with delta and kappa sites comprising 28% and 2%, respectively. In the area adjacent to the central canal, mu sites contributed about 65% of the total opioid sites followed by delta sites at 33% and kappa sites at 2% of total opioid sites. These results demonstrate a differential distribution of mu, delta and kappa binding sites with respect to the organization of the spinal gray matter. The preferential occurrence of all 3 opioid binding sites in the superficial dorsal horn is noteworthy since many fine caliber primary afferent fibers mediating nociception establish synaptic contact in this region.