Karin N. Westlund

Noradrenergic projections to the spinal cord of the rat

Noradrenergic terminals were identified in the spinal cord of rats by immunocytochemical staining for dopamine-beta-hydroxylase. Although immunoreactive fibers and terminals were observed throughout the spinal grey matter, heavier accumulations of terminal labeling were observed in the marginal layer of the dorsal horn, in the ventral horn among motoneurons, and in the autonomic lateral cell columns of the thoracic and sacral spinal cord. Two specific retrograde transport techniques were employed to identify the origins of these noradrenergic terminations in the spinal cord. Cells of origin were observed in the locus coeruleus, the subcoeruleus, the medial and lateral parabrachial, and the Kölliker-Fuse nuclei, as well as adjacent to the superior olivary nucleus. These regions correspond to the A5-A7 cell groups of the pons. No spinally projecting noradrenergic cells were ever observed in the medulla. It was concluded that pontine noradrenergic cell groups are the sole source of noradrenergic terminals in the spinal cord.

Descending serotonergic, peptidergic and cholinergic pathways from the raphe nuclei: A multiple transmitter complex

The localization of serotonergic, various peptidergic and possibly cholinergic neurons in the medullary raphe nuclei that project to the lumbosacral spinal cord have been studied using a retrograde transport method combined with immunocytochemical and histochemical techniques. Spinally projecting neurons stained for serotonin-like, substance P-like, enkephalin-like and thyrotropin-releasing hormone-like immunoreactivity and for the histochemical marker acetylcholinesterase were all observed in each of the raphe nuclei of the medulla, as well as in the adjacent ventrolateral reticular formation. The similar distributions of the descending serotonergic and peptidergic neurons in the raphe nuclei as well as quantitative data on their relative numbers suggest that a large fraction of raphe-spinal neurons contain serotonin co-existing with one or more peptides in the same cell.

Localization of distinct monoamine oxidase a and monoamine oxidase b cell populations in human brainstem

Monoclonal antibodies, specific for either monoamine oxidases A or B, were used to determine the localization of monoamine oxidase in the human brain. Two distinct populations of neurons were detected by immunocytochemical staining. Neurons in regions rich in catecholamines were positive for monoamine oxidase A, including the nucleus locus coeruleus, the nucleus subcoeruleus and the medullary reticular formation. In these regions, monoamine oxidase A could be co-localized with the synthetic enzyme, dopamine-beta-hydroxylase. Neurons in the substantia nigra and the periventricular region of the hypothalamus, areas rich in dopamine neurons, stained for monoamine oxidase A but with much less frequency and intensity. The major accumulation of monoamine oxidase B-positive neurons was observed in the same regions in which monoamine oxidase B is found to co-localize with serotonin in monkey tissues, including the nucleus raphe dorsalis and the nucleus centralis superior. In addition, both monoamine oxidase A and B were localized in distinct populations of neurons in the lateral and tuberal regions of the hypothalamus, a region shown recently to contain histamine neurons in rats. Some glial cells were stained throughout the brain for monoamine oxidase A or B suggesting that glia are capable of either expression or uptake of these proteins.

Origins of serotonergic projections to the spinal cord in rat: An immunocytochemical-retrograde transport study

The origins of the serotonergic projections to the spinal cord in the rat were determined by employing the retrograde cell marker HRP coupled with the unlabeled antibody, peroxidase-antiperoxidase immunocytochemical method of Sternberger. Large numbers of stained neurons (greater than 70%) in the medullary raphe nuclear complex were found to contain both HRP retrogradely transported from the spinal cord and positive 5-HT staining. These serotonergic cell groups, including the nucleus raphe obscurus, raphe pallidus, raphe magnus, and the ventral parts of the reticular formation, project to all spinal cord levels. In addition, some neurons contained HRP granules, but were unstained for 5-HT, suggesting that they may contain other non-serotonergic neurotransmitters. More rostrally in the midbrain reticular formation, many 5-HT neurons were found to have projections exclusively to the cervical spinal cord. These findings indicate that the descending serotonin inputs to the spinal cord originate not only from the serotonergic neurons located in the medullary raphe complex, but also from other new sources located in the central gray and reticular formation of the midbrain.

Excitatory amino acid receptor involvement in peripheral nociceptive transmission in rats

The involvement of excitatory amino acid receptors in peripheral nociceptive processing was assessed in two separate experiments. In the first, one knee joint cavity of rats was injected with 0.1 ml of L-glutamate (0.001 mM; 0.1 mM; 1.0 mM), L-aspartate (0.001 mM; 0.1 mM: 1.0 mM), L-arginine (0.1 mM) or different combinations of these amino acids. The animals tested for paw withdrawal latency to radiant heat and withdrawal threshold to von Frey filaments at different time points. Combinations of glutamate/aspartate, aspartate/arginine or glutamate/aspartate/arginine when injected into the joint, in the absence of any other treatment, reduced the paw withdrawal latency and withdrawal threshold immediately after the injection and persisting up to 5 h indicating the development of hyperalgesia and allodynia. Subsequent intra-articular injection of either an NMDA or a non-NMDA glutamate receptor antagonist ((+/-)-2-amino-7-phosphonoheptanoic acid (AP7), 0.2 mM) or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), 0.1 mM) attenuated the thermal hyperalgesia and the mechanical allodynia produced by glutamate/aspartate/arginine. On the other hand, in a second experiment intra-articular injection of AP7, ketamine or CNQX reversed the hyperalgesia and allodynia produced by injection of a mixture of kaolin and carrageenan into the joint. These receptor antagonists, however, did not have an effect on the joint edema. These findings provide evidence for a potential role of peripheral NMDA and non-NMDA receptors in nociceptive transmission.

Behavioral and immunohistochemical changes in an experimental arthritis model in rats

&NA; An experimental arthritis induced by injection of kaolin and carrageenan into the knee joint resulted in a temporal relationship between glutamate dorsal horn content and paw withdrawal latency (PWL) which was positively correlated. Limping, guarding, increased response to heat stimuli (hyperalgesia) and altered staining patterns for glutamate (GLU), substance P (SP), and calcitonin gene‐related peptide (CGRP) were monitored in the awake behaving arthritic rat over a 1 week time course. A decrease in PWL occurred on the side ipsilateral to the inflamed knee as early as 4 h after the induction of arthritis indicating the animals are hyperalgesic. The PWL remained decreased through the first 24 h. Computer‐assisted quantification of the density of immunohistochemical staining indicated the content of GLU, SP and CGRP was altered differentially throughout the time course of the arthritis. The changes observed for all three substances occurred across the entire superficial dorsal horn. There was an initial depletion of SP followed by an increase in both SP and CGRP content which was maintained through 1 week. The GLU content was increased during the hyperalgesic period. The GLU changes followed the same time course and were positively correlated with the changes in PWL. In a small group of animals injected with kaolin and carrageenan, hyperalgesia did not develop. In this group of animals, no change in dorsal horn GLU or SP content occurred. Rather, there was an increase in CGRP content in the middle portion of the superficial dorsal horn which is the termination site of knee joint afferents. These data indicate that the development of heat hyperalgesia is dependent on GLU and possibly SP. Since inflammation of the knee joint does not involve the foot pad, the heat hyperalgesia observed during the first 24 h following induction of arthritis represents a central neuronal sensitization.

Is there a pathway in the posterior funiculus that signals visceral pain

&NA; The present report provides evidence that axons in the medial part of the posterior column at T10 convey ascending nociceptive signals from pelvic visceral organs. This evidence was obtained from human surgical case studies and histological verification of the lesion in one of these cases, along with neuroanatomical and neurophysiological findings in animal experiments. A restricted lesion in this area can virtually eliminate pelvic pain due to cancer. The results remain excellent even in cases in which somatic structures of the pelvic body wall are involved. Following this procedure, neurological testing reveals no additional neurological deficit. There is no analgesia to pinprick stimuli applied to the body surface, despite the relief of the visceral pain. Since it is reasonable to attribute the favorable results of limited midline myelotomies to the interruption of axons of visceral nociceptive projection neurons in the posterior column, we have performed experiments in rats to test this hypothesis. The results in rats indicate that the dorsal column does indeed include a nociceptive component that signals pelvic visceral pain. The pathway includes neurons of the postsynaptic dorsal column pathway at the L6‐S1 segmental level, axons of these neurons in the fasciculus gracilis, and neurons of the nucleus gracilis and the ventral posterolateral nucleus of the thalamus.

Amino acid release into the knee joint: key role in nociception and inflammation.

&NA; This study examined the release of several amino acids after induction of knee joint inflammation in rats using kaolin and carrageenan. During the initial 10‐min collection after knee joint injection with the irritants, the concentration of glutamate and the nitric oxide metabolites, arginine and citrulline, doubled. This increase persisted for at least two hours. During the same time period aspartate concentrations remained unchanged. Direct knee joint administration of lidocaine prevented the increases in amino acid concentration measurable by microdialysis probe inserted into the joint. These data suggest the possibility that glutamate may be released by neuronal endings in the joint.

Origins and terminations of descending noradrenergic projections to the spinal cord of monkey

This report describes the distribution of noradrenergic cells in the brainstem and the pattern of terminal varicosities in the spinal cord of monkey using the immunocytochemical localization of dopamine-beta-hydroxylase (DBH). Using two separate and equally reliable techniques, retrograde transport of the antibody to DBH and a double-labeling method, the cells of origin of noradrenergic fibers in the spinal cord have been identified. The results of these studies indicate that 79% of all noradrenergic cells with axons projecting to the spinal cord are located in the nucleus subcoeruleus and nucleus locus coeruleus. Other pontine noradrenergic cell groups contribute the remainder of the fibers to the cord. No medullary cells contribute to the noradrenergic innervation of the spinal cord.

Organization of Descending Serotonergic Projections to the Spinal Cord

Publisher Summary This chapter discusses the organization of descending serotonergic projections to the spinal cord. The indolamine serotonin is believed to have important neurotransmitter and/or neuroregulatory roles in the descending pathways influencing various somatosensory, motor, and autonomic functions within the spinal cord. The autonomic preganglionic neurons of the thoracic spinal cord, as well as the motoneurons of the ventral horn, are also under the influence of descending serotonergic cell groups. Cells in these various motoneuron pools can be inhibited and/or excited by iontophoretic application of serotonin or by electrical stimulation of the descending serotonergic pathways. In recent years, the introduction of immunocytochemistry to neurobiology has provided a very sensitive method for studying the organization of descending serotonergic pathways. In contrast to the previously employed histofluorescence methods with which rapid photodecom-position of the yellow fluorescing cells and terminals was a constant problem, the peroxidase-antiperoxidase (PAP) immunochemical method results in a relatively permanent marker for 5-HT neurons and terminals.