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PubMed1993

Expression of phosphorylated high molecular weight neurofilament protein (NF-H) and vimentin in human developing dorsal root ganglia and spinal cord.

The expression of vimentin and the phosphorylated variant of high molecular weight neurofilament protein (NF-H) was studied in developing human fetal dorsal root ganglia and spinal cord. The technique used for examination of cryosections was double-label fluorescence with monoclonal antibodies. Both proteins were present in the nerve fibres inside the ganglia of 6- and 8-week-old embryos. During further development the expression of vimentin continued to increase in the satellite cells, but was found to be decreasing in the ganglion cells. Phosphorylated NF-H was found in the processes of ganglion cells, as well as in the perikarya at all developmental stages. In the spinal cord of 6- and 8-week-old embryos, phosphorylated NF-H protein was found in the longitudinal fibres of the marginal layer and in processes of the mantle zone; some of the fibres also contained vimentin. Later the co-expression of the two proteins ceased and vimentin was found only in glial and mesenchymal derivatives. Phosphorylated NF-H was located, at all developmental stages, in the axons of both white and grey matter, but not in the neuronal perikarya. The results indicate that phosphorylation of the NF-H in human dorsal root ganglia starts in the perikarya of the ganglion cells while in the ganglion cells of the spinal cord it takes place in the axons.

Antibodies, Monoclonal↗
PubMed1992

Morphometric study of the sensory neuron and peripheral nerve changes induced by chronic cisplatin (DDP) administration in rats.

We performed a morphological, morphometric and toxicological study on the spinal ganglia and peripheral nerves of the rat after chronic administration of cisplatin (cis-dichlorodiammineplatinum II; DDP) with two different schedules. Severe damage of the spinal ganglia neurons was demonstrated with predominant involvement of the nucleus and nucleolus associated with a decrease in the cell size. Morphological and morphometric changes also occurred in the sciatic and peroneal nerves with the features of axonopathy. All these changes were more marked in the group of rats which underwent the most intense DDP treatment and the tissue platinum concentrations were also higher in this group. This experimental model is the first available for chronic DDP administration in which concomitant spinal ganglia and peripheral nerve damage has been confirmed pathologically. Our study supports the hypothesis that DDP-induced peripheral nerve fiber degeneration may result from nuclear and nucleolar changes in the sensory ganglion cell perikaryon.

Animals↗
PubMed1994

Neural spread of Junin virus in intraperitoneally inoculated rats.

On the basis of an already demonstrated Junín virus (JV) neural route after peripheral footpad infection of newborn rats, here we attempted to determine the viral pathway following intraperitoneal inoculation. As from the 2nd week post-infection, neurological disease developed reaching 84% mortality at 30 days. Immunoperoxidase labeling of viral antigen, concomitantly with infectivity assays and histological examination, was carried out in serially harvested samples. Whenever infectivity was detected, whether by viral rescue from coculture or by conventional isolation, viral antigen staining was achieved. Infective JV was present at threshold levels in spleen and liver from days 2 to 10, and in blood from days 5 to 15. In neural tissues, viral antigen was initially disclosed at day 5 in thoracic rachideal ganglia and related spinal cord segments. From day 7 thereafter, the entire spinal cord was involved; at this stage, first evidence of viral infection was found in brain stem, with subsequent spread to other encephalon structures at day 10. According to harvested samples, no significant differences were found in labeled cell percentages at thoracic vs cervical or lumbar levels of spinal cord. In contrasts, greater involvement of cerebral cortex versus brain stem, hippocampus or cerebellum was demonstrated shortly before death. Although JV antigen was overwhelmingly predominant in neurons, no morphological changes were apparent in such cells. Since rachideal spinal ganglia and spinal cord infection invariably preceded viral spread to encephalon, concomitantly with viral clearance from lymphoreticular organs and blood, a neural pathway seems warranted.

Animals↗
PubMed1996

Catechol-O-methyltransferase in rat sensory ganglia and spinal cord.

The localization of catechol-O-methyltransferase immunoreactivity in rat dorsal root ganglia and in the spinal cord and its co-existence with substance P, calcitonin gene-related peptide and fluoride-resistant acid phosphatase in dorsal root ganglion cells was examined with immunohistochemical and histochemical double-staining methods. Analysis of dorsal of dorsal root ganglia at both cervical and lumbar levels revealed catechol-O-methyltransferase immunoreactivity in numerous dorsal root ganglion cells. Double-staining studies showed that catechol-O-methyltransferase and substance P immunoreactivities were located in different cells with a few exceptions, whereas both catechol-O-methyltransferase and calcitonin gene-related peptide immunoreactivities were detected in about 10% of all labeled cells positive for one of the two markers at both levels studied. The great majority of fluoride-resistant alkaline phosphatase-positive cells were also immunoreactive for catechol-O-methyltransferase. Again, no difference was found between cervical and lumbar levels. Catechol-O-methyltransferase immunoreactivity was also found in the neuropil of the dorsal horn of the spinal cord. The staining was most intense in the superficial laminae (I-III) and overlapped partly with substance P and calcitonin gene-related peptide immunoreactivity. Western blotting analysis revealed that soluble catechol-O-methyltransferase was the clearly dominating form of the enzyme in dorsal root ganglia. The distribution pattern of catechol-O-methyltransferase in dorsal horn and sensory neurons suggests that the enzyme may modulate sensory neurotransmission.

Acid Phosphatase↗
PubMed1989

The somato-visceral divergent projections of peripheral processes of substance P-containing spinal ganglionic neurons--tri-labeling study of combining fluorescein tracing with immunocytochemistry.

The chemical nature of the spinal ganglionic neuron with peripheral processes projecting divergently to the somatic and visceral areas, has been identified by means of tri-labeling method of combining fluorescein tracing with immunocytochemistry. 10 rats were used. First, 2 microliters of 2% fast blue(FB) were injected into the left coeliac ganglion. Two days later, 2% nuclear yellow (NY) was injected into left 9-11 intercostal nerves, 1 microliter for each. On the 4th day, the animal was perfused with 10% formalin in 0.1 mol phosphate buffer. The left T9-11 spinal ganglia were removed and cut into sections by cryostat. The sections were observed under fluorescence microscope and photographed. The results showed that there were three kinds of neurons in the spinal ganglia: 1) single FB labeled cells with blue fluorescent cytoplasm accounted for 38.8% of total cells; 2) single NY labeled cells with yellow fluorescent nuclei accounted for 52.7%; 3) FB and NY double-labeled cells, mostly small or medium in size, accounted for 8.5%. Then, the sections containing double-labeled cells were further processed by substance P-demonstrating PAP immunocytochemical staining. The photographs with immunostaining and fluorescein labelings in the same section were compared. We found that the labeling ratio of SP/NY was 1.4%, SP/FB was 7%, and SP/NY + FB was 28.8%. The present study detected not only the convergence of somato-visceral sensation in the spinal ganglia but also the chemical nature of these neurons containing substance P (SP) for the first time. In addition, these results may provide a morphological basis for the mechanisms of referred pain and somato-visceral reflection.

Animals↗
PubMed1982

[Lactate dehydrogenase activity and isoenzyme spectrum in the transplanted sensory and sympathetic ganglia of the rat].

Total activity, isoenzyme pattern of lactate dehydrogenase (LDH), morphological peculiarities of normal and transplanted in the mesentery for 1-7 and 30 days dorsal root and superior cervical sympathetic ganglia of the adult rats, were analysed. About 30% neurons in sensory ganglia and 5% in sympathetic ganglia were viable 30 days after transplantation. Total LDH activity was 3 times higher in normal sympathetic ganglia compared with that in spinal ganglia. Transplantation leads to a slight rising in total LDH activity in sensory root ganglia and to a slight fall in sympathetic ganglia. Five fractions of LDH in normal sensory ganglia and four fractions in sympathetic ganglia were revealed. Significant rising of M4 isoenzyme activity was characteristic of both sensory and sympathetic ganglia after transplantation for 7 days. Functional significance of the changes observed is discussed.

Anaerobiosis↗
PubMed1998

Localization and regulation of the delta-opioid receptor in dorsal root ganglia and spinal cord of the rat and monkey: evidence for association with the membrane of large dense-core vesicles.

Using immunohistochemistry and immunoelectron microscopy, the localization and regulation of delta-opioid receptor-like immunoreactivity were studied in dorsal root ganglia and spinal cord of normal rat and monkey, and after peripheral axotomy. Delta-opioid receptor-like immunoreactivity was observed in many small dorsal root ganglion neurons, and in the rat most of them contained substance P and calcitonin gene-related peptide. At the ultrastructural level, delta-opioid receptor-like immunoreactivity was localized in the Golgi complex, on the membrane of the large dense-core vesicles and on the membrane of and/or inside a type of large vesicle with an interior of low electron density. The latter vesicles were often in contact with multivesicular bodies. In the superficial dorsal horn of the spinal cord, most delta-opioid receptor-positive nerve fibers contain substance P and/or calcitonin gene-related peptide, both in rat and monkey. Also, in these nerve endings delta-opioid receptor-like immunoreactivity was found on the membrane of large dense-core vesicles and on the membrane of, or in, the lucent vesicles. Occasionally, delta-opioid receptor-like immunoreactivity was observed on the plasmalemma of the terminals, particularly when the vesicles were in exocytotic contact with the plasmalemma. Peripheral axotomy induced a decrease in delta-opioid receptor-like immunoreactivity both in cell bodies in the dorsal root ganglia and in terminals in the dorsal horn. These data suggest that the delta-opioid receptor may be a constituent of the membrane of large dense-core vesicles storing and releasing neuropeptides. It is suggested that upon exocytotic release of substance P and calcitonin gene-related peptide from large dense-core vesicles, there is a transient modification of the surface of the primary afferent terminals which leads to exposure of the receptor protein so that enkephalin released from adjacent terminals can activate the receptor. The decrease in delta-opioid receptors after axotomy indicates that delta-opioid receptor-mediated inhibitory effects are attenuated at the spinal level both in the rat and monkey.

Animals↗
PubMed1995

Distribution of the extended family of protein kinase C isoenzymes in fetal organs of mice: an immunohistochemical study.

Using isoenzyme-specific antisera, we have studied the distribution of protein kinase C isoforms in fetal mouse organs at the developmental age of 17 days. Two different sets of antibodies, produced by different manufacturers, were employed in this study. The specificity of the antisera was tested by immunoblotting experiments using whole fetal mouse extracts. Immunohistochemistry was carried out by means of an alkaline phosphatase-conjugated secondary antibody. Analysis of fetal mouse longitudinal cryostat sections stained with the antibodies demonstrated a distinct distribution of protein kinase C isoforms in the tissues. Protein kinase C-alpha and C-beta I were present in all tissues examined, whereas the C-beta II isoform was absent in the lung and the liver. Protein kinase C-gamma was identified in brain, spinal ganglia, and adrenal gland. The C-epsilon isoenzyme was abundantly expressed in spinal ganglia and in the smooth muscle cells of the bronchial wall. Antisera to C-zeta and C-eta isoforms heavily stained liver, kidney, and spinal ganglia, whereas the C-theta isozyme was mainly detected in brain, stomach and kidney. Thus, protein kinase C-alpha, C-beta I, C-beta II, C-zeta, C-eta and C-theta were the isoforms present in many of the organs investigated. The two sets of antibodies gave slightly different results that might be ascribed to the different epitopes recognized by the antisera. One set of antisera was employed to investigate the distribution of the isoforms in selected organs from an earlier developmental age (15 days) and from adult animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed2000

Changes in nerve growth factor levels in dorsal root ganglia and spinal nerves in a rat neuropathic pain model.

The purpose of this study was to measure the changes in levels of nerve growth factor (NGF) in dorsal root ganglia (DRG) and spinal nerves with the aim of investigating the role of NGF in a rat neuropathic pain model. Nerve injuries were made by tight ligation of the left L5 and L6 spinal nerves using 6-0 silk thread in male Sprague-Dawley rats. Before surgery and 1, 3, 5, 7, and 14 days after surgery, tissue samples collected included the L3-6 DRGs bilaterally, segments of the ipsilateral L5-6 spinal nerves proximal and distal to ligation sites, and corresponding sites of the contralateral L3-6 and the ipsilateral L3-4 spinal nerves. NGF levels in the DRGs of the injured spinal nerves (the left L5 and L6) did not change significantly from control values. The spinal nerve segments distal to ligation sites had higher levels of NGF than the control values. Unlesioned sites did not show any significant changes in NGF levels. The increase of NGF in distal segments of injured spinal nerves may be due to an accumulation of retrogradely transported NGF. The maintenance of NGF levels in the DRGs that had lost peripheral connections may reflect local synthesis after nerve injury.

Animals↗
PubMed1983

[Effect of ischemia and postischemic circulatory recovery on the ultrastructure of neurons].

In the investigation performed, a comparative analysis on the effect of ischemia and postischemic recirculation upon the neuron in the rat cerebral cortex and the dog spinal ganglia, common regularities and differences in appearance of pathological ultrastructural alterations were revealed. A general tendency towards increasing gravity of the pathological alterations was noted, as the period of ischemia and recirculation grew large. A leading type of the changes in the group of the injured neurons both in the spinal ganglia and in the cerebral cortex were changes of a chromatolitic type and they made one more general regularity. Hyperchromatosis was presented by an essentially less number of cases. The differences observed were presented as a rather greater polymorphism in the cortical neurons alterations comparing to the spinal ganglia cells at ischemia and postischemic recirculation. Besides, in the cortical neurons, more sensitive to the ischemic factor, diverced forms of the compensatory-adaptive reactions were revealed, that demonstrated plastic potentials of the cortical neurons.

Animals↗
PubMed1987

Growth hormone-releasing factor (GRF)-like immunoreactivity in sensory ganglia of the rat.

Growth hormone-releasing factor (GRF)-like immunoreactivity has been demonstrated in the trigeminal and spinal ganglia of fetal, young and adult rats by use of peroxidase-antiperoxidase immunohistochemistry. GRF-like-immunoreactive cells first appear during the second half of embryonic life, as early as day 17. In untreated animals the GRF-immunoreactive elements form approximately 1% of all ganglion cells in the trigeminal and spinal ganglia; their numbers do not change significantly during development. The granular immunoreaction product is confined to perikarya, especially to the perinuclear region. Nerve fibers displaying GRF-like immunoreactivity were found neither in the ganglia, nor in the corresponding central and peripheral areas of termination. The possible role of GRF in sensory ganglia is discussed.

Age Factors↗
PubMed1989

[Intracytoplasmic eosinophilic inclusions in dorsal root ganglia and spinal nerve roots from an autopsy case of unusual familial ataxia with cerebrospinal fluid abnormality].

Studies were performed on the light and electron microscopic structures of the dorsal root ganglia (DRG) and spinal nerve roots of the 4th lumbar nerve obtained by autopsy from a 49-year-old man with unusual familial ataxia, who showed varied neurological manifestations such as progressive ataxia, action tremors, pyramidal tract signs, mild deep sensory disturbances and autonomic dysfunctions during a 30-year period of illness, and had 2 siblings, one male and female, similarly affected and close consanguineous marriages in his family. On laboratory examinations, blood chemistry disclosed no significant findings. Repeated spinal taps showed constant xanthochromia and elevated protein in the cerebrospinal fluids. A PEG and cranial CTs revealed a progressive brain atrophy. NCVs and EMGs in the extremities were within normal limits. There was no chromosomal abnormality. Light microscopically, intracytoplasmic eosinophilic inclusions (IEIs) with pale rim, which showed varied sizes and rounded shapes, occurred within neurons in the DRG, particularly in small neurons. Many of the small neurons had numerous IEIs, and several rounded granules with a high degree of eosinophilia, measuring below 5 microns in diameter. Generally the small neurons showed atrophic, while most large neurons showed no remarkable change although they had a small number of IEIs and granules located in the perikaryal periphery. Most satellite cells, and some Schwann cells in the DRG, ventral and dorsal roots had IEIs similar to those seen in the neurons. No IEIs occurred intraaxonally, and there was seen no degenerative process in the DRG and roots except a connective tissue fiber proliferation in the DRG.(ABSTRACT TRUNCATED AT 250 WORDS)

Ataxia↗
PubMed1985

The origins of innervation of the esophagus of the dog.

This study defined the origins of extrinsic efferent and afferent innervation of the normal canine esophagus. When all the layers of the wall of the 3 esophageal regions (cervical, thoracic and abdominal) were injected with horseradish peroxidase (HRP), labeled nerve cells were found in the nucleus ambiguus (NA) and parasympathetic nucleus of X (PX) of the brainstem. Most labeled cells in the NA were located in the compact column (retrofacial nucleus) while labeled cells in the PX were located in separate rostral and caudal areas. There was no somatotopic organization in either the NA or PX. Labeled sympathetic postganglionic neurons were found in the cranial cervical, middle cervical, cervicothoracic, thoracic sympathetic trunk and celiacomesenteric ganglia. The HRP injection of the esophageal wall labeled sensory cell bodies in the glossopharyngeal, proximal and distal vagal, and C2-T6 spinal ganglia. There was no discernible pattern of distribution of labeled cells in the autonomic or sensory ganglia. When the HRP injections were confined to the mucosa-submucosa layers of the thoracic esophagus, a small number of labeled cells were identified in the NA; however, no labeled cells were found in the NA when injections were confined to the mucosa-submucosa of either the cervical or abdominal esophageal regions. With these confined injections, the labeled nerve cells appeared in the rostral part of the PX. Thus, it appeared that the internal tunics of the esophagus (i.e., the mucosa and submucosa) were innervated by neurons in the rostral PX while the muscular tunic was innervated by neurons in the caudal PX and the rostral NA. After mucosa-submucosa injections, labeled sympathetic neurons appeared in the same ganglia that were identified after whole wall injections and these had a similar random distribution. These injections also labeled neurons in the glossopharyngeal, proximal vagal, and distal vagal ganglia, but unlike the whole wall injections there was no labeling in the spinal ganglia. This suggested that the labeled cells of the spinal ganglia seen after whole wall injections conveyed impulses from the tunica muscularis and serosa.

Animals↗