Dorsal Root Ganglia Of Animals Research Articles

IL-1β Is Involved with the Generation of Pain in Experimental Autoimmune Encephalomyelitis.

Pain is one of the main symptoms of multiple sclerosis, a demyelinating disease of the central nervous system that affects millions of people worldwide. The experimental autoimmune encephalomyelitis (EAE) is considered an experimental model of multiple sclerosis, and besides motor weakness, hypernociception is one of the clinical signs of animals with EAE. In this study, we investigated the influence of some cytokines in the generation of the hypernociceptive response in a mouse model of EAE using MOG35-55. We measured some cytokines in the dorsal root ganglia (DRG), an important anatomical structure involved in pain. We found increased levels of the cytokines TNF-α, IL-1β, and Kc in DRGs of animals with EAE. We used the antibody IL-1ra to antagonize the effects of IL-1β, and animals presented a decrease in the hypernociceptive response. Thus, our results suggest that hypernociception in this experimental model of EAE may be a consequence of the increase in some cytokines in DRGs, especially IL-1β.

Painful Nerve Injury Decreases Resting Cytosolic Calcium Concentrations in Sensory Neurons of Rats

Neuropathic pain is difficult to treat and poorly understood at the cellular level. Although cytoplasmic calcium ([Ca]c) critically regulates neuronal function, the effects of peripheral nerve injury on resting sensory neuronal [Ca]c are unknown. Resting [Ca]c was determined by microfluorometry in Fura-2 AM-loaded neurons dissociated from dorsal root ganglia of animals with hyperalgesia to mechanical stimulation after spinal nerve ligation and section (SNL) at the fifth and sixth lumbar (L5 and L6) levels and from animals after skin incision alone (control group). Axotomized neurons from the L5 dorsal root ganglia were examined separately from adjacent L4 neurons that share the sciatic nerve with degenerating L5 fibers. After SNL, large (34 mum or larger) neurons from the L4 ganglion showed a 29% decrease in resting [Ca]c, whereas those from the L5 ganglion showed a 54% decrease. Small neurons only showed an effect of injury in the axotomized L5 neurons, in which resting [Ca]c decreased by 30%. A decrease in resting [Ca]c was not seen in neurons isolated from rats in which hyperalgesia did not develop after SNL. In separate experiments, SNL reduced resting [Ca]c in capsaicin-insensitive neurons of the L5 ganglion by 60%, but there was no change in neurons from L4. Resting [Ca]c of capsaicin-sensitive neurons was not affected by injury in either ganglion. SNL injury decreased the proportion of neurons sensitive to capsaicin in the L5 group but increased the proportion in the L4 group. Painful SNL nerve injury depresses resting [Ca]c in sensory neurons. This is most marked in axotomized neurons, especially the large and capsaicin-insensitive neurons presumed to transmit non-nociceptive sensory information.

Pyridoxine (vitamin B6) neurotoxicity: enhancement by protein‐deficient diet

Large doses of pyridoxine cause injury to the primary sensory neurons in trigeminal and dorsal root ganglia of animals and patients subjected to megavitamin therapy. The increased hazard to subjects with reduced renal excretory function has been explored previously. In the present work, the neurotoxicity of pyridoxine for rats was found to be increased by dietary protein deficiency. A mere 3 or 7 days of pretreatment with either of two protein-deficient diets were sufficient to accelerate and intensify the clinical neurological signs and histological lesions from pyridoxine injections. These results are caused, at least in part, by loss of body weight, decreased protein binding in serum and decreased consumption of water and decreased volume of urine, which reduce the urinary losses of the toxicant. The vitamers related to pyridoxine (pyridoxal, pyridoxamine) and the coenzyme (pyridoxal 5-phosphate) did not cause clinical signs or lesions similar to those produced by pyridoxine even when injected in maximum tolerated doses. Neither a protein-deficient diet nor bilateral nephrectomy changed the results with the vitamers.

Painful inflammation-induced increase in mu-opioid receptor binding and G-protein coupling in primary afferent neurons.

Opioids mediate their analgesic effects by activating mu-opioid receptors (MOR) not only within the central nervous system but also on peripheral sensory neurons. The peripheral analgesic effects of opioids are best described under inflammatory conditions (e.g., arthritis). The present study investigated the effects of inflammation on MOR binding and G-protein coupling of full versus partial MOR agonists in dorsal root ganglia (DRG) of primary afferent neurons. Our results show that Freund's complete adjuvant (FCA) unilateral hindpaw inflammation induces a significant up-regulation of MOR binding sites (25 to 47 fmol/mg of protein) on DRG membranes without affecting the affinity of either full or partial MOR agonists. In our immunohistochemical studies, the number of MOR-immunoreactive neurons consistently increased. This increase was mostly caused by small-diameter nociceptive DRG neurons. The full agonist DAMGO induced MOR G-protein coupling in DRG of animals without FCA inflammation (EC50 = 56 nM; relative Emax = 100%). FCA inflammation resulted in significant increases in DAMGO-induced MOR G-protein coupling (EC50 = 29 nM; relative Emax = 145%). The partial agonist buprenorphine hydrochloride (BUP) showed no detectable G-protein coupling in DRG of animals without FCA inflammation; however, partial agonist activity of BUP-induced MOR G-protein coupling was detectable in animals with FCA inflammation (EC50 = 1.6 nM; relative Emax = 82%). In behavioral studies, administration of BUP produced significant antinociception only in inflamed but not in noninflamed paws. These findings show that inflammation causes changes in MOR binding and G-protein coupling in primary afferent neurons. They further underscore the important differences in clinical studies testing peripherally active opioids in inflammatory painful conditions.

Ultrastructural Study of the Alterations in Spinal Ganglion Cells of Rats Chronically Fed on Ethanol

A study was conducted to find the effects of chronic alcohol (EtOh) administration on the rat dorsal root ganglion (DRG) cells in vivo. Morphoquantitative changes of the cytoplasmic organelles in neurons and satellite cells (SC) of lumbar DRG of animals fed with 20 and 40% of EtOH for 6 months were determined at the electron microscopic level. Stereological methods were used to quantitatively evaluate the changes in the neuronal Golgi fields, in the lysosomal system components called dense bodies (DB), in the mitochondria, and in the cytoplasmic perikaryal projections (PP) characteristic of DRG neurons. Prolonged consumption of 20% EtOh was well tolerated by neurons. There were, however, some structural modifications in the studied organelles, and there was a significant increase in the neuronal surface. In SC the number of mitochondria and DB increased significantly. Treatment with 40% EtOH produced massive organelle alterations in both neurons and SC, including disruption of the PP, markedly reducing the neuronal surface area. The architecture of the SC sheath appeared disorganized. The alterations resembled those of senescence, and indicated that a high dose of EtOH (or its metabolites) had a profound disruptive effect on the organelles and on the membrane systems of the DRG cells. The SC of the DRG units from the animals fed with EtOH were the first to show significant morphological alterations. When the architecture of the SC sheath already showed evident signs of disorganization, the neuronal body was just beginning to show morphological damage. These results suggest that the progressive disorganization of the SC sheath is a probable source of complication in peripheral neuropathy.

Cell types and axonal sizes of calcitonin gene-related peptide-containing primary sensory neurons of the rat

Neurons with immunoreactivity (IR) for calcitonin gene-related peptide (CGRP) in the rat dorsal root ganglia were examined by immunoelectron microscopy. About one-half of the neurons of the L 5 dorsal root ganglia of animals treated with colchicine had CGRP-IR, and 40% of these were large neurons of type A. The proximal parts of their peripheral axons were, however, unmyelinated (91%) or thinly myelinated (9%). Thickly myelinated axons observed in the same sections were always devoid of CGRP-IR. The CGRP-IR neurons were various subtypes of type A and B neurons. No specific morphological characteristics were associated with CGRP-IR.

Axoplasmic transport in the toad Bufo marinus

The rate and course of axoplasmic transport from the eighth dorsal root ganglion cell bodies into the sciatic nerve of the toad Bufo marinus were studied. Concentrated tritiated proline was hydraulically injected into a surgically exposed dorsal root ganglion of animals maintained at 19 ± 0.5 °C. At postinjection intervals of 1, 6, and 10 h, the animals were sacrificed and the dorsal root, ganglion, and sciatic nerve were removed bilaterally. The dorsal roots and peripheral nerves were cut into 3 mm segments measured from the ganglion. In some experiments all tissues were prepared for liquid scintillation counting techniques. In others the ganglion and every fifth 3 mm nerve segment were fixed in Bouin's fixative for radioautography, and the remaining tissue segments were prepared for liquid scintillation counting methods. Scintillation counts (counts/min) of consecutive segments along the labeled nerve were plotted against distance (mm) for each animal. Examination of these profiles showed a peak of radioactivity in the injected ganglion for each animal that was followed distally by an abrupt in the adjacent segments of the nerve. Radioactivity remained relatively stable in subsequent segments forming a plateau and then dropped to baseline levels forming a wavefront in the distal portion of the peripheral nerves of the 6 and 10 h toads. Movement of this wavefront during the 6 to 10 h time interval provided evidence for an axoplasmic flow rate of about 120 mm/day. Radioautographs of the ganglion and representative segments along the sciatic nerve were examined with both bright- and dark-field microscopy. Accumulations of silver grains were observed overlying the injected ganglion cell bodies and labeled axons of the nerve. Values for the number of silver grains/unit area were obtained from dark-field radioautographs of the nerve segments through the use of a computer-microscope system. The relative amount of radioactivity present in the axons was thereby demonstrated. Radioautographic data confirmed the axoplasmic flow rate of about 120 mm/day determined by liquid scintillation counting methods for the bulk of radioactive materials transported at this fast rate. In addition, the evaluation of radioautographic data suggests that an even faster flow rate of 185-–215 mm/day may exist for a small portion of the labeled materials transported in the axon.