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A J Beitz

Publications and source records attributed to A J Beitz.

At least 19 recordsLinked to original sources

Induction of NADPH-diaphorase/nitric oxide synthase in the brainstem trigeminal system resulting from cerebellar lesions.

Recent evidence indicates that NADPH-diaphorase (NADPH-d) and nitric oxide synthase (NOS) can be induced in cerebellar afferent neurons following mechanical, thermal, or chemical damage to the cerebellar cortex (Saxon and Beitz [1994] Neuroreport 5:809-812). The present study reports on the induction of NADPH-d/NOS in neurons of the brainstem trigeminal complex (BVC). Three groups of rats were used: Group I received a unilateral glass micropipette lesion into the vermal/paravermal region of the cerebellar cortex, group II received a concurrent injection of fluoro-gold along with the pipette lesion, and in group III the cerebellar cortex on one side was aspirated. Following survival times of 7-120 days, animals were processed for NADPH-d histochemistry. All three groups showed projection-specific induction of NADPH-d in different regions of the brainstem trigeminal complex. Induced neurons were distributed throughout the ipsilateral subnucleus interpolaris, principal trigeminal nucleus, and intertrigeminal nucleus. Subnucleus oralis contained a small number of induced neurons localized to the ipsilateral dorsomedial portion of the subnucleus. Projection-specific induction was confirmed by the presence of neurons double-labeled for NADPH-d and Fluoro-Gold. Although the functional consequences of NADPH-d/NOS induction remain to be elucidated, the induction of these enzymes in precerebellar neurons suggests that nitric oxide may play a role in the neuronal response to target specific lesions.

Animals

Altered c-fos expression in the parabrachial nucleus in a rodent model of CFA-induced peripheral inflammation.

Increases in the expression of immediate early genes have been shown to occur in the lumbar spinal cord dorsal horn after peripheral inflammation. Given that the pontine parabrachial nucleus has been implicated in nociceptive as well as antinociceptive processes and is reciprocally connected with the spinal cord dorsal horn, it seems likely that peripheral inflammation will cause alterations in immediate early gene expression in this nucleus. To test this hypothesis we examined cFos-like immunoreactivity in a rodent complete Freund's adjuvant-induced peripheral inflammatory model of persistent nociception. Unilateral hind paw injections of complete Freund's adjuvant produced inflammation, hyperalgesia of the affected limb, and alterations in open field behaviors. Immunocytochemical analysis demonstrated a bilateral increase in cFos-like immunoreactivity in the lateral and Kolliker-Fuse subdivisions of the parabrachial nucleus at 6 and 24 hours postinjection and an ipsilateral decrease below basal levels in the Kolliker-Fuse subdivision at 96 hours postinjection when compared to saline controls. Taken together, these results suggest that select parabrachial neurons are activated by noxious somatic inflammation. These active parabrachial neurons are likely to participate in ascending nociceptive and/or descending antinociceptive pathways.

Animals

Spinal cord NMDA receptors modulate peripheral immune responses and spinal cord c-fos expression after immune challenge in rats subjected to unilateral mononeuropathy.

To characterize further the neural involvement in local immune reactions, we evaluated the effect of intrathecal NMDA-receptor blocker dizocilpine maleate (MK-801) on the peripheral immune response itself and on spinal cord c-fos expression induced by the delayed-type hypersensitivity (DTH) response. Immune challenge took place in the hind paw ipsilateral or contralateral to an injured sciatic nerve in both previously sensitized and immune-naive animals. An enhanced immune response was observed bilaterally in the hind paws of animals subjected to unilateral mononeuropathy compared with sham-operated controls. In contrast, no such enhancement was observed when neuropathic animals were challenged in the front paws. The increased DTH response was blocked successfully by the intrathecal administration of an analgesic dose of MK-801. Compared with sham-operated animals, animals subjected to unilateral mononeuropathy showed both a differential distribution and an increase in the number of c-fos-labeled neurons in the dorsal horn of the L3-L5 spinal cord segments after immune challenge. This was observed irrespective of whether the challenge took place ipsilateral or contralateral to the injured nerve. In addition to reversing the changes in immune response, intrathecal administration of MK-801 reversed the pattern of c-fos immunoreactivity in the spinal cord after immune challenge in neuropathic animals. These data suggest that select groups of spinal cord neurons participate in enhancing the peripheral immune response to a specific antigen in neuropathic animals and that this enhancement involves central NMDA receptors.

Animals

Increased delayed type hypersensitivity in rats subjected to unilateral mononeuropathy is mediated by neurokinin-1 receptors.

An animal model or peripheral mononeuropathy was utilized in the present study to investigate the potential role of substance P (SP) in modifying immune responses associated with chronic pain conditions. Animals subjected to unilateral sciatic ligation and sham-operated animals were sensitized with keyhole limpet hemocyanin (KLH) and subsequently challenged in the ipsilateral or contralateral hind paw to produce a delayed-type hypersensitivity (DTH) response. Subcutaneous microdialysis and radioimmunoassay were used to measure interstitial fluid SP levels in the challenged tissue prior to and following immune challenge in control and neuropathic animals. Following immune challenge, there was a significant increase in the concentration of SP in tissue dialysate samples from the challenged paw of both sham-operated and neuropathic animals. However, tissue SP levels in neuropathic animals were more than two-fold higher than those obtained from sham-operated controls following challenge. SP concentration remained elevated for 2.5 h following immune challenge in neuropathic animals compared to 90 min in sham-operated animals. Compared with controls, neuropathic animals also exhibited an increased DTH response that was reversed, in a dose-related fashion, by the non-peptide NK-1 receptor blocker L-703,606. The same antagonist had no effect in sham-operated animals. These data suggest that the increased DTH response in animals subjected to unilateral mononeuropathy involves SP and NK-1 receptors present in the challenged tissue.

Animals

An experimental model for the non-invasive trans-synaptic induction of nitric oxide synthase in Purkinje cells of the rat cerebellum.

Nitric oxide synthase and its product nitric oxide are believed to be significantly involved in neuronal degeneration associated with various excitotoxic events and in some acute and chronic neuropathological disease states. The synchronous activation of inferior olive neurons by the drug harmaline is known to produce tremorogenic activity in rats via increased stimulation of the olivocerebellar system. Following subcutaneous or intraperitoneal injection (15, 30 or 60 mg/kg) of harmaline and survival intervals of three days or more, NADPH-diaphorase histochemistry and nitric oxide synthase immunocytochemistry revealed nitric oxide synthase induction in Purkinje cells of the cerebellar vermis and paravermis, but not the lateral hemispheres. Double labeling with NADPH-diaphorase and calbindin revealed that induced Purkinje cells contain calbindin and that induced Purkinje cells are not necessarily associated with regions of Purkinje cell degeneration. The induction of nitric oxide synthase in a subpopulation of Purkinje cells may reflect a protective response of selected Purkinje cells to overstimulation of specific olivocerebellar projections. This is the first experimental model for the non-invasive induction of nitric oxide synthase in central nervous system neurons and as such may provide a useful tool for studying the induction of nitric oxide synthase associated with excitotoxic events.

Amino Acid Sequence

Electrical stimulation of the sciatic nerve alters neuropeptide content and lymphocyte migration in the subcutaneous tissue of the rat hind paw.

To study the possible mechanism by which peripheral nerves mediate immune responses in target tissues, electrical stimulation of the sciatic nerve was combined with subcutaneous microdialysis of the hind paw. Following unilateral stimulation of the sciatic nerve, an ipsilateral rise in substance P and a bilateral rise in VIP levels were observed in dialysate samples from experimental vs control animals. Electrical stimulation of the sciatic nerve induced a marked hyperemia and swelling of the ipsilateral paw. Quantitative immunocytochemical analysis of paraffin-embedded sections of the hind foot pads demonstrated T lymphocyte migration ipsilateral to the stimulated nerve. These findings suggest that peripheral nerves can directly modulate local immune and inflammatory responses.

Animals

Basal release of Met-enkephalin and neurotensin in the ventrolateral periaqueductal gray matter of the rat: a microdialysis study of antinociceptive circuits.

The periaqueductal gray (PAG) contains neural circuits that participate in descending antinociception. Anatomical and electrophysiological evidence suggests that these circuits might employ opioid peptides and GABA in series to remove a tonic inhibition of descending PAG output neurons. The present studies examined the release of the antinociceptive peptides Met-enkephalin and neurotensin in the ventrolateral PAG, and investigated the interaction between GABA and Met-enkephalin release. In awake and freely moving rats the ventrolateral PAG was dialysed using 25 ga. concentric probes. Basal release of peptide in 12 min or 40 min fractions was determined using radioimmunoassays. To establish how the ventrolateral PAG responds to nociception, dialysis was performed following unilateral hindpaw inflammation using Complete Freund's Adjuvant. Twenty-four hours after inflammation was induced, neurotensin release was increased 133% and Met-enkephalin release was increased 353% compared to control animals. Seven days after inflammation was induced, neurotensin release declined precipitously, while basal Met-enkephalin release remained elevated 313% above controls. Thus, unlike enkephalin, increased basal neurotensin release is not sustained with persistent tonic nociception. In addition, we confirmed in normal animals that the ventrolateral PAG is induced to release Met-enkephalin by systemic morphine. A 43% increase in basal Met-enkephalin release was observed immediately following a 12 mg/kg i.p. morphine injection. Morphine should have the opposite effect (inhibit peptide release) if it acts directly on the enkephalinergic neurons. Thus, we examined the hypothesis that GABAergic interneurons in the PAG mediated morphine-stimulated enkephalin release. When the GABAantagonist bicuculline (0.25 microM to 25 microM) was co-infused with the dialysis medium, Met-enkephalin release increased in a dose-dependent fashion and peaked 68% above pre-infusion levels. These data elucidate the reciprocal inhibitory relationship between GABA and enkephalin in the ventrolateral PAG. We hypothesize that, when nociception induces Met-enkephalin release within this region, the tonic GABAergic inhibition is overcome, resulting in greater sensitivity of PAG enkephalinergic neurons. Ultimately, this enhanced enkephalin release should result in greater excitability of the descending PAG output neurons that are responsible for antinociception.

Animals

NMDA R1 mRNA distribution in motor and thalamic-projecting sensory neurons in the rat spinal cord and brain stem.

The N-methyl-D-aspartate (NMDA) receptor is important in both sensory and motor neurotransmission. In this study we examine NMDA R1 mRNA hybridization signal over individual sensory and motor neurons in the spinal cord and brain stem. A significantly greater quantity of NMDA R1 mRNA was present in motor neurons of the lumbar spinal cord and hypoglossal nucleus compared to thalamic projecting sensory neurons in the spinal cord dorsal horn, the spinal trigeminal nucleus pars caudalis and the cuneate and gracile nuclei. No significant difference in the quantity of NMDA R1 mRNA was observed between sensory neurons known to relay predominantly nociceptive information (trigeminothalamic and spinothalamic tract neurons) and that relay predominantly touch and proprioceptive information (dorsal column neurons).

Animals

Castration increases [125I]MK801 binding in the hippocampus of male rats.

This study examines the effect of castration and androgen replacement on [125I]MK801 binding in the hippocampus. In castrated male rats, [125I]MK801 binding was significantly increased in both the stratum oriens and radiatum and the pyramidal cell layer of CA1. In contrast, no increase in [125I]MK801 binding was observed in the stratum oriens and radiatum of CA1 of castrated rats that were treated with dihydrotestosterone. No change in [125I]MK801 binding was observed in the CA3 region or dentate gyrus after castration. The observed increase in [125I]MK801 binding in pyramidal cell neurons within CA1 suggests that androgens may potentially affect hippocampal function by modulating pyramidal cell NMDA receptors.

Androgens

Distribution of NMDAR1 receptor subunit mRNA and [125I]MK-801 binding in the hypothalamus of intact, castrate and castrate-DHTP treated male rats.

This study examines NMDAR1 receptor subunit mRNA expression and [125I]MK-801 binding in hypothalamic and limbic nuclei of intact, castrate and castrate-dihydrotestosterone propionate (DHTP)-treated male rats. In intact rats, the highest levels of NMDAR1 mRNA were observed in the supraoptic, suprachiasmatic, ventromedial and arcuate nuclei. Low levels of hybridization were observed in the bed nucleus of the stria terminalis, lateral preoptic area, lateral hypothalamic area and lateral septum. In castrated rats both NMDAR1 mRNA and [125I]MK-801 binding are significantly decreased in the lateral septum compared to castrate rats treated with DHTP, a non-aromatizable androgen. NMDAR1 mRNA was also significantly decreased in the supraoptic nucleus of castrate rats when compared to castrate rats treated with DHTP. These data suggest that androgens may modulate NMDA receptor function in some parts of the central nervous system.

Animals

N-methyl-D-aspartate R1 messenger RNA and [125I]MK-801 binding decrease in rat spinal cord after unilateral hind paw inflammation.

Recent evidence suggests that N-methyl-D-aspartate receptors play an important role in the etiology and maintenance of chronic nociception. Previous studies have demonstrated that tissue injury or stimulation of nociceptive afferent projections results in the expansion of receptive fields, hyperalgesia and C-fiber-induced wind-up, events that can be inhibited by N-methyl-D-aspartate antagonists. This study examines the effect of unilateral hind paw inflammation on N-methyl-D-aspartate R1 messenger RNA and [125I]dizocilpine maleate binding in the L4-L5 segments of the lumbar spinal cord of rats. Spinal cords were examined at 7.5 h, three, seven and 20 days after injection of the left hind paw with 120 microliters of complete Freund's adjuvant. N-methyl-D-aspartate R1 messenger RNA, as measured with in situ hybridization, was observed to decrease bilaterally in laminae I, II and X of the lumbar spinal cord. This decrease was evident in laminae I and II at 7.5 h and three days after hind paw injection. In lamina X, a postinjection decrease in hybridization signal was observed at 7.5 h and seven days. A bilateral decrease in [125I]dizocilpine maleate binding was observed in laminae I and II at three, seven and 20 days after paw injection. This observed decrease in binding at the N-methyl-D-aspartate receptor suggests a compensatory mechanism by which N-methyl-D-aspartate-mediated nociceptive events may be modulated.

Animals

Immunocytochemical evidence that quisqualate is selectively internalized into a subset of hippocampal neurons.

Quisqualic acid (QUIS) has been shown to interact with several glutamate receptor subtypes and uptake sites. We have previously demonstrated that a brief exposure of hippocampal cells to QUIS sensitizes them to depolarization by the alpha-amino-omega-phosphonate analogues of glutamate, AP4, AP5, and AP6. This QUIS-induced sensitization is accompanied by the active uptake of QUIS into hippocampal slices. In order to localize the sites of QUIS uptake into rat hippocampal slices, a polyclonal antibody against QUIS was raised in rabbits. Utilizing immunocytochemical techniques, we have identified immunoreactive axons and dendrites after brief exposure times to QUIS, and perikarya after longer exposure times to QUIS. The intensity of the QUIS immunoreactivity increased as the exposure time to QUIS increased. QUIS immunoreactivity was primarily found in stratum oriens and stratum radiatum, of regions CA1, CA2, and CA3 of the hippocampus as well as in the hilus and molecular layer of the dentate gyrus. The distribution and morphology of QUIS immunoreactive cells appeared to be similar to those of GABAergic interneurons. Glial fibrillary acidic protein (GFAP) did not co-localize with the QUIS-internalizing cells suggesting that they are not glia. Ultrastructural analysis revealed QUIS immunoreactive profiles within the stratum radiatum. Immunostained profiles at both the light and EM levels appeared, in many cases, to be swollen and showed signs of degeneration. Such changes were only evident in tissue exposed to QUIS. These data demonstrate that QUIS is taken up by a select group of neurons in the rat hippocampus.

Animals

Differential NMDA NR1 mRNA expression among spinal trigeminal neurons that project to different targets.

The N-methyl-D-aspartate (NMDA) NR1 glutamate receptor subtype has been proposed to play an important role in the transmission of orofacial sensory information in the spinal trigeminal nucleus (STN). The distribution of NR1 mRNA expression in the STN and its relationship to STN projection neurons has not been investigated previously. Using neuroanatomical tract tracing with in situ hybridization techniques, we found that neurons in the STN that project to the thalamus, cerebellum and spinal cord expressed more mRNA for NR1 than do nonprojection neurons. Trigeminothalamic neurons were found to express more NR1 mRNA than trigeminospinal or trigeminocerebellar neurons. Thus, NMDA-specific excitatory amino acids may be more efficacious in the relay of orofacial information to the thalamus than to the spinal cord or cerebellum.

Animals

Distribution of nitric oxide synthase-immunoreactive interneurons in the spinal trigeminal nucleus.

The spinal trigeminal nucleus is involved in the transmission of orofacial sensory information. Neither the distribution of the neuromessenger, nitric oxide, within the trigeminal system nor the possible relationship of this simple gas with trigeminothalamic neurons has been carefully studied. Using immunocytochemical (against nitric oxide synthase) and histochemical (NADPH-diaphorase staining) techniques, we have found that nitric oxide neurons and processes are more prominent in the nucleus caudalis and the dorsomedial aspect of the nucleus oralis than in other spinal trigeminal regions. To study the relationship of nitric oxide to trigeminothalamic neurons and intertrigeminal interneurons of the spinal trigeminal nucleus, spinal trigeminal neurons were retrogradely labeled with fluorogold by thalamic injections or by injections into the junction of the nucleus interpolaris and nucleus caudalis. Medullary sections were subsequently processed with NADPH-diaphorase histochemistry. None of the diaphorase-stained neurons in the spinal trigeminal nucleus was found to contain fluorogold; however, some diaphorase-stained processes were found in close proximity to trigeminothalamic neurons. Following spinal trigeminal nucleus injections, many diaphorase-stained neurons were found to contain fluorogold, especially in the nucleus caudalis, suggesting that nitric oxide-containing neurons in the spinal trigeminal nucleus are intertrigeminal interneurons. Collectively, these data indicate that nitric oxide is most prominent in interneurons located in nucleus caudalis and that these interneurons give rise to processes that appose trigeminothalamic neurons, raising the possibility that they may indirectly influence orofacial nociceptive processing at the level of the spinal trigeminal nucleus via nitric oxide production.

Amino Acid Oxidoreductases

NMDA receptor mRNA expression in NOS-containing neurons in the spinal trigeminal nucleus of the rat.

The spinal trigeminal nucleus (STN) is involved in the transmission of orofacial sensory information. Nitric oxide (NO), an important neuromessenger, and the glutamate receptor subtype, NMDA NR1, have been implicated in nociception in the STN. However, the anatomical relationship of NO and NMDA NR1 has not been investigated within this nucleus. Using both immunocytochemical (against NO synthase; NOS) and in situ hybridization studies of NMDA NR1 receptor mRNA, we found that NOS-containing neurons in the STN expressed more mRNA for NR1 than did non-NOS-containing neurons in the STN. These data suggest that NMDA activation may lead to NO production in the STN and is consistent with previous studies, implicating both NMDA and NO in nociception.

Amino Acid Oxidoreductases

Characterization of the effect of cholecystokinin (CCK) on neurons in the periaqueductal gray of the rat: immunocytochemical and in vivo and in vitro electrophysiological studies.

The periaqueductal gray (PAG) is an important integration site for pain, autonomic functions, vocalization, fear and anxiety. Cholecystokinin (CCK) is a major neurotransmitter in the PAG and CCK receptors are heterogeneously distributed within the PAG. Since CCK antagonists are anxiolytic and potentiate morphine analgesia, it is possible that these effects of CCK are mediated through alteration of neuronal activities in the PAG. The goals of this study were to examine the anatomical and physiological properties of the PAG CCK containing systems. The distribution of CCK-containing axons and boutons in PAG was examined using immunohistochemical procedures. These studies show that CCK-like immunoreactive (CCK-LIR) fibers and terminals are present throughout PAG, but are particularly heavily concentrated in a focal column that runs longitudinally throughout the rostrocaudal axis of dorsolateral PAG and in nucleus cuneiformis which represents a caudolateral extension of PAG. The physiological effects of CCK on PAG neurons were examined in both in vivo and in vitro preparations. In the in vivo experiments multibarreled electrodes were used to record from PAG neurons and to apply CCK and the CCK antagonists, CR1409 and proglumide. Of 37 neurons recorded in vivo, CCK caused excitation in 25 cells, inhibited 7 cells and had no effect on 5 cells. The excitatory effect was blocked by CR1409 in 11/11 cells tested. Proglumide blocked the excitatory response of CCK in 12/14 cells. Proglumide blocked the inhibitory effect in 2 of 7 cells, but CR1409 had no effect on CCK-evoked inhibition in 7 cells tested. Extracellular, conventional intracellular and whole cell patch clamping procedures were used to study CCK actions in the in vitro slice preparation. In the extracellular recording experiments, responses of PAG cells to CCK were measured in slices that were maintained at 22 degrees C (room temperature) and at 32 degrees C. CCK excited 40/56, inhibited 7/56 and had no effect on 9/56 cells; excitatory responses were blocked by CR1409 in 32/36 cells and by proglumide in 25/27 cells tested. Inhibitory responses to CCK were unaffected by CR1409, but were blocked in 3/7 cells by proglumide. Conventional intracellular recordings were made from 13 cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Cerebellar injury induces NOS in Purkinje cells and cerebellar afferent neurons.

Purkinje cells of the cerebellar cortex and neurons of most precerebellar nuclei are conspicuous by the absence of constitutive (neuronal) nitrix oxide synthase (nNOS). Here we show that following mechanical, chemical or thermal injury to the cerebellar cortex there is an induction of nNOS in Purkinje cells as identified with NADPH-diaphorase (NADPH-d) histochemistry and nNOS immunocytochemistry. Induced nNOS and NADPH-d (inNOS/NADPH-d) first appeared 72 hours post-treatment and persisted in excess of 8 weeks. Precerebellar neurons throughout the brain stem also exhibited induced nNOS and NADPH-d with a similar time course. These results indicate that brain lesions can induce nNOS in local neurons and in neurons which are afferent to the leasioned area.

Amino Acid Oxidoreductases

Hemilabyrinthectomy causes both an increase and a decrease in corticotropin releasing factor mRNA in rat inferior olive.

It was previously shown [NeuroReport, 3 (1992) 829-832] that unilateral labyrinthectomy (UL) induces Fos expression in several brainstem regions, including the beta subnucleus of the inferior olive. Using isotopic 33P in situ hybridization, the present results demonstrate significant changes in oligonucleotide-probed mRNA levels for corticotropin-releasing factor (CRF) in the rat inferior olivary nucleus 4 days following unilateral labyrinthectomy (UL). In the medulla of normal animals there was strong CRF mRNA labeling in the inferior olivary nucleus, and weaker labeling in the vestibular nuclei and prepositus hypoglossi. Following unilateral labyrinthectomy, the contralateral olivary beta subnucleus showed a significant increase in CRF message, similar to the contralateral Fos labeling observed after hemilabyrinthectomy [NeuroReport, 3 (1992) 829-832]. In addition, the contralateral A and B subnuclei (IOA/B) of the inferior olive showed a strong increase in CRF labeling, while the ipsilateral dorsal cap of Kooy (IOK) showed a decrease. This novel bidirectional alteration in CRF message in different subdivisions of the same nuclear group indicates the existence of both up and down regulatory mechanisms controlling CRF peptide expression, and reflects the dynamic neurochemical alterations occurring during vestibular compensation.

Animals