Release of nerve growth factor by human glial cells in culture.
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Adolescent↗
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Results for “GANGLIA, SPINAL”
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Substrate requirement and media supplements necessary for the long-term survival of chick sympathetic and sensory neurons cultured without serum.
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Animals↗
Promotion of survival and neurite outgrowth of cultured peripheral neurons by exogenous lipids and detergents.
Gangliosides, in particular the monosialoglycosphingolipids Gtet 1 (GM1), have previously been implicated in the mediation of neuronal rescue and restitutional axonal growth, both in vitro and subsequent to brain and peripheral nerve lesions. In the present study it is shown that the bis-sialosyl gangliosides Gtet2b and Gtet3b, but not the gangliosides Gtet2a and Gtet1, promote the survival of dissociated dorsal root ganglion (DRG) neurons cultured from Embryonic Day (E) 8 chicks (DRG8) almost to the same extent as nerve growth factor (NGF). Ciliary ganglion (CG) neurons from E8 chicks (CG8) and DRG10 neurons were virtually not supported suggesting considerable specificity in terms of neuronal targets and developmental stages being addressed. Moreover, a variety of other lipids including cerebroside (Cb), dipalmitoylphosphatidylcholine (DPPC) and -serine (DPPS), sulfatide (Sf), and sphingomyelin (Sm) were tested for putative survival promoting activity toward chick CG, DRG, and lumbar sympathetic ganglion (SG11) neurons. At the highest concentration employed (2.5 x 10(-5) M), Sm, DPPC, and DPPS maintained between 45 and 65% of the plateau survival with CG8 (maximally supported by ciliary neuronotrophic factor (CNTF], DRG8, and DRG10 neurons, and 30 to 40% with SG11 neurons. Cb supported CG8 neurons at about 55% of the plateau value achieved with CNTF, but had hardly any effect on the other neuron populations tested. Control experiments using highly enriched neurons and serum-free conditions assured that the effects were unlikely to be mediated by serum components or nonneuronal cells. A variety of detergents, in particular Triton X-100, also promoted the survival of CG8 and DRG10 neurons. Ganglioside Gtet1, Sm, and Triton X-100 shifted the NGF titration curve for DRG10 neurons between 6- and 15-fold in a dose-dependent manner suggesting synergisms between NGF and lipids for neuronal maintenance. These results document the neuronotrophic potency of certain gangliosides, a heterogeneous group of structurally unrelated lipids, and detergents. The mechanisms by which these agents modulate neuronal survival still await clarification.
Animals↗
Changes in synthesis of specific proteins in axotomized dorsal root ganglia.
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Animals↗
Alterations of [3H]actinomycin D binding to axotomized dorsal root ganglion cell nuclei: an autoradiographic method to detect changes in chromatin structure and RNA synthesis.
An autoradiographic method was developed to quantify on a comparative basis the binding of [3H]actinomycin D (Act D) to the cell nuclei of frozen, unfixed sections of spinal sensory ganglia in rats. After a crush lesion of the sciatic nerve, alterations of [3H]Act D binding were found in L5 and L6 dorsal root ganglia which corresponded to changes in RNA synthesis observed in other studies. An increase in Act D binding was seen at 1 to 3 days postoperation, followed by a decrease at 5 to 7 days. By 9 to 11 days a second increase in binding occurred, followed by a decrease at 14 days. Contralateral ganglia exhibited an increase in Act D binding only at 5 days compared with unoperated controls. The timing of the response in axotomized ganglia differed with the distance of the lesion from the cell body. The observed patterns of Act D binding confirm that changes of chromatin structure are closely associated with the alterations of RNA and protein synthesis occurring after axon injury. The method may be useful as an indicator for alterations in RNA synthesis related to changes in chromatin structure in complex tissues.
Animals↗
Sensory and sympathetic ganglia in HIV-1 infection: immunocytochemical demonstration of HIV-1 viral antigens, increased MHC class II antigen expression and mild reactive inflammation.
Sensory and sympathetic ganglia from 12 cases of human immunodeficiency virus type 1 (HIV-1) infection, all but one without clinical evidence of peripheral nerve disease, were studied immunocytochemically for their content of lymphocytes, macrophages, MHC Class II antigens and HIV-1 and cytomegalovirus antigens. They were compared with ganglia from 7 normal and peripheral nerve disease control cases. Compared with normal controls, many of the ganglia from the majority of HIV-1-infected subjects contained more T lymphocytes and macrophages and enhanced MHC class II expression. A few also showed occasional neuronal degeneration which was not present in the normal controls. In 7 cases HIV-1 gp41 envelope protein and/or p24 core protein antigens were detected in intraganglionic macrophages. Sensory ganglia contained more gp41 HIV-1 antigen than sympathetic ganglia. There was no clear correlation between detection of HIV-1 antigens in ganglia and in the CNS. Detection of HIV-1 antigens in ganglia was more common in cases of HIV-1 infection that had progressed to clinical AIDS by the time of death (71%) than in those that had not done so (40%). It is concluded that there is commonly a mild ganglionitis which is asymptomatic in the absence of detailed clinical testing and frequently associated with local presence of HIV-1 antigens in sensory and sympathetic ganglia in AIDS.
Acquired Immunodeficiency Syndrome↗
Cell adhesion mechanisms in gangliogenesis studied in avian embryo and in a model system.
Neural crest cells undergo rapid changes in their cell-to-cell and cell-to-extracellular matrix adhesion during the ontogeny of the peripheral nervous system. The mechanisms of adhesion have been analyzed to assess the respective roles played by the cell adhesion molecules (CAMs) and the differentiated junctions. Crest cells which lose their terminal bar junctions after emigration from the neural tube contain only very few gap junctions during gangliogenesis. The calcium-dependent cell adhesion molecules, L-CAM, disappear from the neural crest and never reappear in crest cell derivatives. In contrast, the number of calcium-independent cell adhesion molecules, N-CAM, diminishes transiently during the migratory phase. In vitro, N-CAM is expressed de novo either just before or at the onset of aggregation into autonomic ganglion rudiments, whereas it is delayed in the dorsal root ganglion cells. In vitro, N-CAM mediates the calcium-independent aggregation mechanism; the rate of aggregation is, however, similar whether crest cells are derived from well-spread cultures or from two- and three-dimensional clusters. Crest cells also exhibit a calcium-dependent mechanism of adhesion controlled by molecules differing from N-CAM but which may codistribute on many different cell types during embryogenesis. These two classes of cell adhesion molecules are present on the surface of neural precursors prior to their differentiation into neurons and glial cells.
Animals↗
Familial amyloidotic polyneuropathy type 1 in Kumamoto, Japan: a clinicopathologic, histochemical, immunohistochemical, and ultrastructural study.
Seventeen autopsy and five biopsy cases of familial amyloidotic polyneuropathy were examined clinicopathologically, histochemically, immunohistochemically, and ultrastructurally. In the autopsy cases, amyloid deposits were predominant in the peripheral nerve tissues, autonomic nervous system, choroid plexus, cardiovascular system, and kidneys. Amyloid involvements in the anterior and posterior roots of the spinal cord, spinal ganglia, thyroid, and gastrointestinal tract were also frequent. In the cardiac conduction system, amyloid deposition was prominent in the sinoatrial node and in limbs of the intraventricular bundle. In the sural nerve biopsy, besides amyloid deposits, degenerative changes of nerve fibers and Schwann cells were detected ultrastructurally, and the morphometric analysis showed a marked reduction in the number of myelinated fibers which correlated with the clinical stage. Amyloid deposits were resistant to pretreatment with potassium permanganate in Congo red staining, and transthyretin was confirmed immunohistochemically as a major component of amyloid deposits, along with the presence of serum amyloid P-component. Besides the amyloid deposits, transthyretin was proven in the liver cells, epithelial cells of the choroid plexus, and pancreatic islet A cells, suggesting that the transthyretin produced by these cells is secreted, transferred into tissues, and deposited in situ as the major component of amyloid in this disorder.
Adult↗
Light and electron microscopic features of peripheral ganglion cells cultured from young and aged Wistar rats.
Neurons of the dorsal root ganglion (DRG) and sympathetic superior cervical ganglion (SCG) were cultured as explants from young adult (3 months old) and aged (28 months old) Wistar rats. Both aged DRG and SCG neurons showed delayed neurite outgrowth compared to young adult neurons. Young and some aged cultured neurons had an ultrastructure similar to that found in normal uncultured cells, but most of the aged cultured neurons displayed a heavy accumulation of homogenous lipid-like inclusions in addition to classic age pigments. Occasionally, large neurofilament aggregates were seen in aged DRG neurons. They were sometimes localized perinuclearly, resembling neurofibrillary tangles. The results show that even very old peripheral neurons survive in culture. As aged cultured neurons show degenerative changes not observed in young adult neurons, it is suggested that cultured peripheral neurons of different ages could provide useful means for neuronal aging studies.
Aging↗
Protein kinase C-beta-like immunoreactivity and phosphorylation dependent immunoreactivity of neurofilaments in developing, adult and aged rat peripheral neurons.
The localizations of protein kinase C-beta-like immunoreactivity (PKC-beta-LI) and phosphorylated neurofilament-like immunoreactivity (PNF-LI) were studied in developing (E12-P28), adult (3-month-old) and aged (15- and 30-month-old) rat dorsal root (DRG) and superior cervical (SCG) ganglia. A close correlation was found between the localization of PKC-beta-LI and PNF-LI in all age groups. PKC-beta-LI was already present in the rat DRG at the time of the appearance of PNF-LI (E12) and after E15 both PKC-beta-and PNF-LI were found in the same subpopulation of neuronal perikarya. Aging had no effect on the distribution or intensity of either PKC-beta or PNF-LI. In the SCG neurons no association was observed between immunoreactivity and accumulation of lipopigments, but in the DRG neurons, lipopigment accumulation was seen in PKC-beta and PNF-immunoreactive large neurons. The results suggest that PKC-beta may regulate the time of appearance and phosphorylation state of some forms of phosphorylated neurofilaments during development. The results also show that no major change occurs in the amount or distribution of PNFs during normal aging.
Aging↗
Mice lacking nerve growth factor display perinatal loss of sensory and sympathetic neurons yet develop basal forebrain cholinergic neurons.
Homologous recombination was utilized to generate mice with a deletion in the coding sequence of the nerve growth factor (NGF) gene. Animals homozygous for NGF disruption failed to respond to noxious mechanical stimuli, and histological analysis revealed profound cell loss in both sensory and sympathetic ganglia. Within dorsal root ganglia, effects of the mutation appeared to be restricted to small and medium peptidergic neurons. These observations confirm the critical dependence of sensory and sympathetic neurons on NGF and demonstrate that other neurotrophins are not able to compensate for the loss of NGF action on these cells. Examination of the central nervous system revealed that, in marked contrast with neurons of sensory and sympathetic ganglia, basal forebrain cholinergic neurons differentiate and continue to express phenotypic markers for the life span of the null mutant mice. Thus, differentiation and initial survival of central NGF-responsive neurons can occur in the absence of NGF.
Animals↗
An in vitro bioassay for neurite growth using cryostat sections of nervous tissue as a substratum.
An in vitro bioassay is described that can be used for studying neurite growth, cell adhesion, and cell migration, as well as other cellular behaviors. The bioassay, which uses tissue sections as substrata for either dissociated cell preparations or explants, offers several distinct advantages over other commonly used bioassays. In particular, this bioassay approximates in vivo cellular environments, and the preparation of large numbers of tissue sections as substrata is relatively straight forward and simple. The method can be employed for studying cell behavior on different ages or types of tissue, as a bioassay for screening antibodies for their ability to perturb a particular cellular behavior, and for assessing the biological activities of growth factors absorbed to specific tissue environments. We present a detailed description of the method, including the variety of options to be considered at each step in the procedure, with an emphasis on using the assay to study neurite growth.
Animals↗
Immunohistochemical studies on enteric substance P of extrinsic origin in the cat.
The origin of substance P (SP)-like immunoreactivity in the Auerbach's plexuses of the esophagus, stomach duodenum was examined in the cat. The intensity of the fluorescence and the area of distribution of SP-like immunoreactivity in the Auerbach's plexuses of the cardiac part of the esophagus and the corpus of the stomach were markedly reduced by bilateral ligation of the abdominal vagus nerve, while little change occurred in the antrum of the stomach and the duodenum. After splanchnic nerve ligation or celiac ganglionectomy, the SP-like immunoreactivity in the Auerbach's plexuses of the antrum of the stomach and the duodenum was markedly reduced, whereas that in the esophagus and the corpus of the stomach was only slightly attenuated. Intense SP-like immunoreactivity was observed in the portions of the abdominal vagus and splanchnic nerve cranial to the ligation. This the Auerbach's plexuses of the esophagus, stomach and duodenum were considered to contain SP transported through the vagus and splanchnic nerve. It was also indicated experimentally that the SP-like immunoreactive fibers in the auerbach's plexus arose from the nodose and spinal ganglia as well as from the dorsal vagus nucleus in the medulla oblongata.
Animals↗
Afferent innervation of the lower oesophageal sphincter of the cat. An HRP study.
Labeling of afferent neurons by the retrograde axonal transport of horseradish peroxidase (HRP) was performed on anaesthetized cats in order to examine the afferent innervation of the lower oesophageal sphincter (LOS), involving both the vagal and the sympathetic nerves. The labeled cells, whose fibres follow the sympathetic pathways were found in dorsal root ganglia from T1 to L2. Nerve section experiments indicated that the main pathways involved were the splanchnic nerves, as expected from classical data. Additional pathways passing through the sympathetic cardiac branch emerging from the stellate ganglion and the thoracic sympathetic branches were also evidenced. This work corroborated the electrophysiological data showing the richness of the LOS sensory vagal innervation. Nevertheless, in this case the difficulties related to the HRP technique are particularly enhanced since the abdominal sensory vagal fibres can be affected by HRP injections.
Animals↗
Localization of sympathetic and sensory neurons innervating the rat kidney.
Following injection of horseradish peroxidase (HRP) into the hilar region of the left kidney of the rat, 66% of labeled sympathetic neurons were located in the ipsilateral paravertebral ganglia, with most cells in T13 and L1, and 14% were located in equivalent segments of the contralateral chain. A similar distribution of sympathetic neurons projected to the right kidney, with most cells in T12 and T13 paravertebral ganglia. Only 20% of the total sympathetic supply to either kidney arose from the prevertebral ganglia. The renal sensory innervation was also bilateral in origin, with about 80% of the neurons arising from ipsilateral dorsal root ganglia. Injection of HRP into the caudal and rostral poles of the left kidney labeled paravertebral neurons which were concentrated in ganglia L1 and T13, respectively, but did not label any sensory neurons. We conclude that most of the renal sympathetic innervation is paravertebral in origin, and that a substantial bilateral component exists for both sympathetic and sensory supplies. Neurons arising from the contralateral side have their cell bodies in segments that provide the main ipsilateral innervation to the same kidney. The majority of sensory axons appear to be restricted to subcortical areas.
Adrenergic Fibers↗
Carotid chemosensory timing effects on cervical sympathetic discharges in the cat.
The hypothesis that respiratory phase-related sympathetic nerve activity would manifest timing effect of carotid chemosensory input with respect to the central respiratory drive was tested in the anesthetized, paralyzed and artificially ventilated cats which were also vagotomized and tracheostomized. Preganglionic cervical sympathetic nerve fibers (PSNF) which were clearly responsive to carotid chemosensory stimulation were selected for the test. Simultaneously with the PSNF, phrenic nerve (PN) and internal intercostal expiratory nerve (IICEN) activities were also recorded. In most instances, carotid chemosensory discharges were monitored in order to register the precise timing of its input to the brain-stem. Timed stimulation of carotid chemoreceptors was elicited by bolus injections of cyanide (10-20 micrograms), nicotine (10-20 micrograms) and CO2-saturated saline (0.2-0.5 ml) into the base of the common carotid artery. Two types of PSNFs were identified: type I discharged only in synchrony with the PN activity (13/47) and type II fired independently of, but entrained to, PN activity (34/47). Type I displayed characteristic timing effects of carotid chemoreflex on PN discharges. However, the sympathetic activity did not share the after-discharge of PN which persisted beyond the carotid chemoreceptor stimulation. Type II did not manifest any timing effect of carotid chemoreflex with respect to PN and IICEN activities. These results suggest the following model of carotid chemoreflex effects on sympathetic neuron activities: a group is exclusively gated by the properties of central respiratory drive whereas the other is not gated but entrained by it in the spontaneously breathing anesthetized cat.
Anesthesia↗
The ovarian innervation in the dog: a preliminary study for the base for electro-acupuncture.
The origin of the canine ovarian sensory and sympathetic nerves was studied by applying horseradish peroxidase (HRP) or wheat germ agglutinin conjugated to HRP (WGA-HRP) to the ovarian stroma and into the ovarian bursa. HRP/WGA-HRP positive neurons were found bilaterally in the dorsal root ganglia of T10 to L4 segment with the majority located in T13 to L2. In sympathetic paravertebral ganglia, labeled neurons were distributed bilaterally in ganglia from T11 to L4 with the majorities located in segments T13 to L2. Both distributions show ipsilateral predominance. Labeled prevertebral neurons were mainly located in the aorticorenal ganglion, ovarian ganglia and caudal mesenteric ganglion. No labeled neurons were found in the dorsal motor nucleus of vagus, nodose ganglia or sacral segment from S1 to S3. This study provides the possible morphological basis of electro-acupuncture concerning the somato-visceral reflex of the ovary.
Animals↗
Localization of NADPH diaphorase in bladder afferent and postganglionic efferent neurons of the rat.
NADPH diaphorase histochemistry was used in combination with axonal labelling techniques to determine if NADPH diaphorase is present in afferent and postganglionic efferent pathways to the urinary bladder of the rat. In the L6 and S1 dorsal root ganglia, 80.9 and 78.5%, respectively, of bladder afferent neurons labelled with fluorescent dyes were NADPH diaphorase positive. In the major pelvic ganglion (MPG), many non-labelled neurons and fibers were intensely stained for NADPH diaphorase. Intensely stained cells were clustered near the exit of the penile nerve although stained cells were also scattered throughout the ganglion. Only a small percentage (3.5%) of bladder postganglionic neurons in the MPG were NADPH diaphorase positive. Since NADPH diaphorase activity commonly reflects the presence of nitric oxide synthase, the present findings raise the possibility that nitric oxide may have a role as a neurotransmitter or neuromodulator in afferent pathways from the urinary bladder.
Amino Acid Oxidoreductases↗