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Isolation of Herpes simplex virus from human trigeminal ganglia, including ganglia from one patient with multiple sclerosis.

Herpes-simplex virus (H.S.V.) was isolated from 18 of 39 trigeminal ganglia (T.G.) obtained within 12 h of death. The virus was isolated from ten persons who had died of trauma, from one case of lymphoma, and from one case of multiple sclerosis. In the cadaver with histologically confirmed multiple sclerosis, large bilateral areas of demyelination were present near the points of entry of the nerve root, and the possibility that H.S.V. migration to the root entry zone caused demyelination cannot be excluded.

Adolescent

Modification and regeneration of synaptic connections in cultured leech ganglia.

Segmental ganglia of the central nervous system of the leech were maintained in culture medium outside the animal for several weeks in order to study the properties of synapses and regeneration by identified sensory and motor nerve cells. A variety of preparations were used, including single ganglia, chains of ganglia and ganglia connected to the areas of skin and muscle that they normally innervate in the animal. (1) For up to 10 weeks after removal from the animal, resting and action potentials recorded from sensory and motor neurons resembled those seen in normal ganglia. The same individual cell in a cultured ganglion could be recorded from with intracellular electrodes on a second occasion after an interval of a few days. (2) Sensory cells, identified as touch, pressure or nociceptive according to their morphology and electrical properties, continued to respond selectively to stimuli of the appropriate modality applied to their receptive fields in the skin; action potentials in motor cells caused contractions in the appropriate muscles. Culture of ganglia for more than 3 weeks caused the disappearance of synaptic potentials and a loss of transparency in ganglia. (3) Certain chemically mediated synaptic interactions between sensory and motor nerve cells became markedly changed in cultured ganglia. These changes appeared over the first 3 weeks and consisted of abnormally large excitatory and inhibitory synaptic potentials. The changes in synaptic transmission observed in culture were in many respects similar to those occurring in ganglia maintained within an animal after lesions have been made in the nervous system (Jansen et al. 1974). (4) The morphological appearances of sensory cells were compared in cultured and normal ganglia after injection of horseradish peroxidase. In cultured ganglia, the branching pattern appeared normal, but varicosities became more conspicuous. (5) When connectives linking cultured ganglia were crushed or cut, regeneration occurred. By 7 days, impulses propagated through the regenerated fibers and evoked synaptic potentials on cells within the next ganglion. The course taken by regenerating axons was observed in cells injected with horseradish peroxidase. The results again resembled those seen in animals with similar lesions. (6) The cultured ganglia provide preparations in which it is possible to analyze the mechanisms that underlie long-term changes similar to those seen in the leech central nervous system in situ.

Animals

Organ culture of rat superior cervical ganglia.

The maintenance of adrenergic function has been investigated in organ cultures of adult rat superior cervical ganglia. Tyrosine hydroxylase and monoamine oxidase activities decreased gradually through 72 hours of culture. Dopamine beta-hydroxylase activity gradually increased in the cultured ganglia to 150% of that seen in fresh ganglia after 14 hours in culture. The level of norepinephrine increased in the cultured ganglia to a maximum of 225% of that seen in control ganglia. The increases in both dopamine beta-hydroxylase activity and in norepinephrine levels required protein synthesis but they were not dependent on the presence of nerve growth factor. The accumulation of norepinephrine in the cultured ganglia could not be explained by an increased rate of amine synthesis. Turnover measurements suggest that the accumulation can be explained by a decreased rate of amine metabolism in the cultured ganglia. A comparison of the distribution of amine storage vesicles showed that the number of heavy vesicles was greater in cultured than in fresh ganglia. The results suggest that sympathetic ganglia develop an increased capacity for amine storage in culture and that this leads to an increase in intracellular levels of norepinephrine.

Animals

Herpesvirus in sensory and autonomic ganglia after eye infection.

In herpesvirus hominis (HVH) infections, virus harbored in the sensory ganglia is now thought to be the main source of recurrent infection at peripheral sites. Experimental HVH infection of the external eye in rabbits produces an acute infection and then latent infection of the trigeminal ganglion. In this study, acute infection of the automatic ganglia serving the eye (superior cervical and ciliary) as well as trigeminal ganglia occurred after HVH inoculation of rabbits' corneas with a herpes type 1 strain (RE). Latent virus infection was detected in the trigeminal ganglion of one of five animals tested six months after initial infection. Since the superior cervical and other autonomic ganglia serving the eye become infected during acute herpes simplex virus infection of the external eye in rabbits, it is possible that these ganglia are also sources of reinfection in recurrent herpetic disease of the eye. Following the initial eye disease with this virus strain, HVH shedding could not be demonstrated even after induction attempts by topically applied epinephrine or systemic use of cyclophosphamide. Thus, establishment of latent HVH infection in the ganglia and chronic shedding of virus into the external eye is not a constant feature of this animal model, but may depend on the specific strain of herpesvirus used.

Animals

On the occurrence of substance P-containing fibers in sympathetic ganglia: immunohistochemical evidence.

Several sympathetic ganglia of the guinea pig, cat and rat were studied with indirect immunofluorescence technique using antibodies to substance P (SP) and dopamine-beta-hydroxylase (DBH), the latter of which represents a marker for noradrenaline containing neurons. In all ganglia studied SP-like immunoreactivity could be observed in certain nerve fibers (SP-positive fibers). Their number, localization fluorescence intensity and 'fluorescence morphology' varied considerably between the different ganglia as well as between species. In the inferior mesenteric ganglion and the coeliac-superior mesenteric ganglion complex of all species, and in particular of the guinea pig, a dense plexus of varicose SP-positive fibers was observed around the mostly DBH-positive, principal ganglion cells. In contrast no such fibers were seen in relation to the DBH-positive SIF cells of the ganglia. No SP-positive ganglion cells were observed in any of the ganglia studied. Previous biochemical and immunohistochemical studies have indicated that peripheral SP-positive fibers may represent the branches of primary sensory neurons. The rich supply of varicose, SP-positive nerve terminals, often surrounding the noradrenergic ganglion cells in a basket-like manner, may indicate that these fibers may play a functional role within the ganglia. Instead of merely passing through the ganglion they may be involved in special types of reflex arches.

Animals

Identification of small intensely fluorescent (SIF) cells as chromaffin cells in bullfrog sympathetic ganglia.

Small intensely fluoresent (SIF) cells were investigated in the ninth and tenth paravertebral sympathetic ganglia of the bullfrog using histochemical and electron microscopic techniques. Fluorescence histochemistry revealed that clusters of SIF cells are sparsely distributed in the ninth and tenth ganglia; the clusters were usually located in the vicinity of blood vessels. Fluorescent processes were not observed emanating from SIF cells. The clusters stained positively for the chromaffin reaction indicating that SIF cells are chromaffin cells. Ultrastructurally, the SIF-chromaffin cells in the sympathetic ganglia appeared virtually identical to the chromaffin cells in the adrenal gland; this includes two cell types that appear morphologically the same as the epinephrine and norepinephrine containing chromaffin cells in the adrenal gland. Efferent synapses from the SIF-chromaffin cells in sympathetic ganglia to sympathetic ganglia to sympathetic neurons were not observed. The SIF-chromaffin cells in bullfrog sympathetic ganglia did not have the morphological characteristics of interneurons; it is suggested that they may function as extra-adrenal chromaffin tissue.

Adrenal Glands

A study of the distribution of chromaffin-positive (CH+) and small intensely fluorescent (SIF) cells in sympathetic ganglia of the rat at various ages.

Small intensely fluorescent (SIF) and chromaffin-positive (CH+) cells were independently investigated by formol-induced fluorescence and by chromaffin techniques in the superior cervical, thoracic and coeliac-mesenteric ganglia of neonatal (2--10 days), adolescent (2--4 months) and adult (6--15 months) rats. Identification of CH+ cells was facilitated by glutaraldehyde fixation prior to chromatin. Intraganglionic blood vessels were displaced by antemortem injection of either India ink or the fluorescent dye Thioflavine-S. SIF and CH+ cells were randomly distributed through the ganglia, either singly or in pairs related to principal neurons, or in variably-sized, highly vascularized groups. In chromaffin preparations these groups either consisted entirely of CH+ cells or else they contained a mixture of CH+ and CH- cells. CH+ cells were present in some adolescent and adult ganglia of all types, and in the neonatal coeliac-mesenteric ganglion at 10 days. In neonatal material generally, SIF cells were mostly green fluorescent, occurring separately or in homogeneous or mixed groups, but both yellow and green cells occurred in coeliac-mesenteric ganglia at 10 days. All ganglia in adolescent and older animals contained both yellow and green cells. There were more green than yellow cells, and more SIF than CH+ cells in all ganglia studied.

Age Factors

Ultrastructure and biochemistry of sympathetic ganglia in idiopathic orthostatic hypotension.

The role of transsynaptic dysfunction in the pathogenesis of idiopathic orthostatic hypotension (IOH, or idiopathic autonomic insufficiency) was examined microscopically and biochemically in autopsy specimens. Light microscopy of the sympathetic ganglia showed abnormalities in all 4 IOH patients, including focal phagocytosis of neurons, increased numbers of satellite cells, and perivascular lymphocytic infiltrates. Electron microscopy revealed proliferation and hypertrophy of satellite cells and abnormalities in the unmyelinated axons. In contrast, the spinal cord intermediolateral columns, containing the presynaptic neurons, were unremarkable in 1 patient, exhibited only mild gliosis in another, and showed neuron loss and fibrillary gliosis in 2 patients. Postsynaptic dopamine-beta-hydroxylase (DBH) activity was decreased at least fourfold (p less than 0.02) in sympathetic ganglia of patients with IOH, while tyrosine hydroxylase (T-OH) was normal. Ganglion choline acetyltransferase (ChAc) activity, an index of presynaptic function and integrity, was normal in the IOH group. A number of our observations suggest that presynaptic disease is not an absolute requirement for adrenergic abnormalities in IOH. The intermediolateral columns of the spinal cord were histologically normal in 2 of the patients with IOH, and ultrastructural abnormalities in sympathetic ganglia were consistent with primary adrenergic degeneration. In addition, presynaptic ChAc activity was normal in IOH ganglia, whereas postsynaptic DBH activity was depressed. Finally, postsynaptic T-OH activity, which is regulated by transsynaptic mechanisms, was normal in IOH ganglia.

Adrenergic Fibers

Norepinephrine turnover in pineal gland and superior cervical ganglia. Changes after gonadotrophin administration to castrated rats.

The effects of FSH and LH on norepinephrine (NE) turnover in pineal gland and superior cervical ganglia of castrated female rats were examined by measuring the decline of [3H]NE concentration in the organ 30--120 min after a single i.v. injection of the labeled amine. Both FSH and LH treatment significantly decreased [3H]NE turnover in the pineal gland whereas only FSH accelerated it in the ganglia. As compared to controls, the initial uptake of [3H]NE was lower in the pineal gland of FSH- and LH-treated animals, and higher in the superior cervical ganglia of rats injected with FSH. Neither FSH nor LH modified [3H]NE turnover in heart or adrenal gland, nor changed their endogenous catecholamine content. Pineal monoamine oxidase (MAO) activity type B (assaying by using beta-phenylethylamine as substrate) was decreased by FSH or LH injection. Only FSH modified MAO activity in the ganglia by increasing significantly type A enzyme (assaying by using serotonin as substrate). These results suggest that FSH and LH affect significantly NE metabolism in pineal gland and superior cervical ganglia of spayed rats.

Adrenal Glands

The polypeptide and the DNA restriction enzyme profiles of spontaneous isolates of herpes simplex virus type 1 from explants of human trigeminal, superior cervical and vagus ganglia.

Analysis of the infected cell polypeptides and the DNA restriction profiles of 31 HSV-1 isolates from the trigeminal, superior cervical and vagus ganglia from 17 individuals (12 U.S.A., 2 Japanese, 3 Norwegian) could be classified as 15 different virus strains. With the exception of the three Norwegian isolates which gave identical profiles, virus isolates from the ganglia of different individuals could all be distinguished from one another. In contrast virus isolates from the trigeminal, superior cervical and vagus ganglia of the same individual, or virus isolates from the left and right ganglia of the same individual or multiple isolates from different explants of a single ganglion were indistinguishable. In conclusion, a single virus strain infects each individual initially and virus descended from this event subsequently infects and becomes latent in different cells of the same ganglion as well as in different ganglia.

DNA Restriction Enzymes

Reflex pathways in the abdominal prevertebral ganglia: evidence for a colo-colonic inhibitory reflex.

1. In vitro experiments were performed on preparations consisting of prevertebral ganglia attached to the entire colon of guinea-pigs. The colon was divided into an orad and a caudad segment and intraluminal pressure was recorded from the terminal end of each segment. Intracellular recordings were simultaneously obtained from neurones in the coeliac plexus. 2. The source of mechanosensory input from the colon paralleled the responses to mesenteric nerve stimulation. That is, section of the mesenteric nerve that contributed the strongest synaptic input to a neurone eliminated most of the mechanosensory input to that neurone. 3. The origin of the mechanosensory input to some neurones could be localized as coming from either the orad or caudad segment of the colon. In the coeliac ganglia 68% of the neurones tested responded primarily to orad distension and 37% to caudad distension. In the superior mesenteric ganglion 57% responded to orad distension and 43% to caudad distension. 4. Repetitive stimulation of the mesenteric nerve trunks arising from the prevertebral ganglia inhibited contractions differentially in the orad and caudad segments. The inferior coeliac nerves inhibited primarily the orad segments of colon and the lumbar colonic nerves inhibited primarily the caudad segments of colon. Stimulation of the superior coeliac nerves did not alter the motility of either segment. 5. When one of the colonic segments was distended, contractions in the other colonic segment were inhibited in 71% of the distensions. This inhibition operated in both directions: either orad inhibiting caudad or vice versa. 6. Cutting the intermesenteric nerve which communicates between the orad and caudad prevertebral ganglia eliminated the inhibitory reflex. 7. These experiments provide evidence for a colo-colonic inhibitory reflex mediated through pathways in the prevertebral ganglia.

Afferent Pathways

Barbiturates block calcium uptake by stimulated and potassium-depolarized rat sympathetic ganglia.

The effects of two barbiturates on calcium uptake by sympathetic ganglia have been examined. Sodium pentobarbital (0.4-0.75 mM) and sodium thiopental (0.3 mM) block the preganglionic stimulation-induced uptake of 45Ca by rat superior cervical ganglia but not action potential conduction in the presynaptic axons. The ganglionic-blocking agent, tetraethylammonium, does not inhibit stimulation-induced Ca uptake and does not prevent the blocking effect of thiopental. This effect is therefore probably presynaptic. Postassium-rich media also stimulate Ca uptake by the ganglia, and this effect is markedly inhibited by pentobarbital. Since the K stimulation effect is also observed in deafferented ganglia but not in guanethidine-denervated ganglia, this effect is probably associated primarily with postsynaptic elements. In sum, the data suggest that the barbiturates inhibit Ca permeability changes in both pre- and postsynaptic neurons.

Animals

Biochemical and histofluorescence studies of catecholamines in superior cervical ganglia in organ culture.

The metabolism of catecholamines in rat superior cervical ganglia in organ culture was examined by biochemical and histofluorescence methods. Pronounced increases in both norepinephrine and dopamine content were observed in the cultured ganglia. Norepinephrine levels were more than doubled after 12 hours in culture and reached a maximum after 24 hours. The greatest increase in norepinephrine concentration occurred in the region of the postganglionic nerve trunks and was correlated with an accumulation of intense catecholamine fluorescence in the stumps of the severed postganglionic nerves. Since the rate of norepinephrine synthesis was unaltered, the increase in norepinephrine levels in cultured ganglia appears to be a result of blocked axoplasmic transport of this amine out of the ganglia. The dopamine content of the cultured preparations was not altered after 24 hours but increased rapidly thereafter and attained a maximum at 48 hours. The changes in dopamine did not parallel the changes in norepinephrine either in time course or distribution. The elevated levels of dopamine were accompanied by an increased rate of dopamine synthesis. It is suggested that the increased dopamine content in cultured ganglia is the result of an enhanced synthesis of this amine by specific dopaminergic cells.

Animals

Virology and histopathology of the trigeminal ganglia of Americans and Japanese.

Herpes simplex virus in the trigeminal ganglia of humans was studied in Philadelphia, Pennsylvania, United States of America, and in Kyoto, Japan. The prevalence of recurrent herpes labialis and of clinically latent herpes simplex virus within trigeminal ganglia was determined in inhabitants of the two cities. In addition, a comparison was made of the prevalence of mononuclear cell infiltration in the trigeminal ganglia of Americans and Japanese. Recurrent herpes labialis was found to be significantly less common in the Japanese city than in the American city. Herpes simplex virus was rescued less commonly from the trigeminal ganglia of cadavers in Japan than in America. The difference was significant. The frequency of mononuclear cell infiltration in the trigeminal ganglia of Americans and Japanese is not significantly different. These observations, as well as previously reported serological studies, suggest that despite the ubiquitous nature of herpes simplex virus in America and Japan, the Japanese have less clinically overt disease caused by this virus.

Adolescent

Cochlear communication routes in the guinea pig--spiral ganglia and osseous spiral laminae: an electron microscope study using microsphere tracers.

The microscopic and ultrastructural anatomy of the spiral ganglia and osseous spiral laminae (OSL) of the guinea pig were studied using the transmission and scanning electron microscopes. The nerve and vascular elements within the spiral ganglia and OSL are surrounded by distinct perineural and perivascular spaces maintained by a loose meshwork of cells and connective tissue fibers. The connective tissue cells are fibers within the OSL and spiral ganglia are distinctly different from the arachnoid-like reticular cells and elastin fibers of the modiolus described in an earlier study. The present experiment demonstrates a pathway between the perineural spaces of the modiolus, spiral ganglia, and OSL. The perineural spaces of the modiolus were previously shown to communicate with the subarachnoid space; therefore, perineural communication from the cerebrospinal fluid to the perilymph via this route is likely. Using mircosphere tracers injected intrathecally, this communication route between the subarachnoid space and scala tympani was further defined. The tracer experiment suggests that the perineural spaces within the internal acoustic meatus, modiolus, spiral ganglia, and OSL are normally patent and do not represent artifacts induced during tissue processing.

Animals

Mechanisms of tyrosine hydroxylase and dopamine beta-hydroxylase induction in organ cultures of rat sympathetic ganglia by potassium depolarization and cholinomimetics.

It was the aim of the present study to elucidate the mechanisms involved in specific tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) induction by potassium depolarization and cholinomimetics in rat superior cervical ganglia kept in organ culture. The effect of high (54 mM) potassium concentration on intact ganglia seems to result in a dual action: a) a specific induction of TH and DBH via release of acetylcholine from preganglionic cholinergic nerve terminals. b) a non-specific effect on terminal adrenergic neurons resulting in a general increase of protein synthesis as indicated by the increase in DOPA decarboxylase (DDC) and monoamine oxidase (MAO) activities. In decentralized superior cervical ganglia potassium depolarization failed to produce the specific TH and DBH induction although a small increase in DDC activity persisted. Carbamylcholine, acetylcholine and nicotine at concentrations of 10(-4) M elicited a selective induction of TH and DBH both in intact and decentralized ganglia via nicotinic receptor stimulation. Bethanechol, predominantly stimulating muscarinic receptors had no significant effect on TH activity. A 4 h pulse of 10(-4) M carbamylcholine produced optimal induction of DBH and TH 24 h and 48 h later respectively. Longer exposure to carbamylcholine resulted in a significantly smaller rise in TH activity.

Acetylcholine