Neurohistochemical study of autonomic nerve ganglia and lungs in burns

Adreno-cholinergic nerve structures of vegetative nerve ganglia and lungs in 20 departed in different periods of burn disease aged from 19 to 85 have been studied with neurohistological methods. Definite dynamics of mentioned structure changes have been shown. It was stated that in different stages of burn disease adreno-cholinergic nerve structures of vegetative nerve ganglia and lung tissues changed greatly and afferent part suffered more in comparison with efferent one. Adrenergic fluorescence was considerably weaker in elderly and senile patients in comparison with younger ones. Luminescence deterioration was more noticeable in neuron cytoplasm. In young aged such nerve structures were more reactive and remarkable with more compensating abilities.     

Pathomorphology of the nodose ganglia of the vagus nerves in burns

Pathomorphology of ganglion nodosum was studied by means of neurohistological methods in 65 dead patients during different periods of burn disease and demonstrated a definite dynamic of structural modifications. In period of burn shock there were discovered dystrophic changes and necrosis of neuron separate groups, varicose thickness of a myelin sheath, in period of toxemia hypertrophied neurons with thick axons predominated, in period of septic toxemia and burn exhaustion predominated atrophic changes, necrotic neurons and grain-blockage disintegration of nervous fibres.     

Pathomorphology of stellate ganglia in thermal injuries of the body

Structural changes of stellate-ganglia in 80 patients aged from 20 to 80 dead in different stages of burn disease (shock, toxemia, septico-toxemia and burn exhaustion) were studied with neurohistological and neurohistochemical methods. It was determined that the increasing of neuron's reactivity was the sign of its changes at the early stages of burn disease. Later hypertrophy, atrophy and neuron's body destruction took place. At the period of burn shock excessively bright luminescence sympathetic neurons prevailed, at the period of toxemia its number decreased. At the period of toxemia and septico-toxemia for the first time it was determined the increase of lipofuscin insertion in adrenergic neurons as well as the increase of the activity not only at the shock period but also at the next periods of burn disease.     

Pathomorphology of adrenergic and cholinergic structures of sympathetic nerve ganglia in experimental burn injuries]

Changes in adreno- and cholinergic structures of sympathetic nerve ganglia (superior cervical, stellate, and splanchnic ganglia of the solar plexus) were studied in 15 male white rats aged 5-7 months, b. m. 200-250 g, 3, 7, and 11 days after burn injury (Stages IIIA, IIIB, involving 20-25% of body surface) and in 5 reference animals. The sections were treated in 2% glyoxylic acid solution and by the Karnovsky-Roots technique. Reduced catecholamine concentrations were revealed in sympathetic nerve ganglia neurons in the early periods after burn injury; the mediator reserves are recovered to a certain measure in later periods after thermal injury. The detected shifts in the sympathetic nerve ganglia neurons correlate with the detected shifts in the cardiovascular system.     

The pathomorphology of the superior cervical sympathetic ganglia in burns

Structural changes of upper cervical sympathetic ganglions of 75 patients who died in different stages of burn disease at the age of 19-85, were studied by neurohistological and neurohistochemical methods. It was determined that neurones change at earlier stages of burn disease begins with their reactivity increase. Later hypertrophy, atrophy and decay of neurones body take place. Sharp cateholamine exhaustion in the structures of upper cervical sympathetic ganglions is also determined during the first days of burn trauma (burn shock stage). Some restoring of mediator stock takes place in subsequent periods of burn disease.     

Dynamics of the retrograde transport of horseradish peroxidase in the trigeminal nerve during burn keratitis

The dynamics of trigeminal neuron labelling with horseradish peroxidase (HRP) has been studied in rats during thermal burn keratitis. HRP application on the cornea after burn prolongs the time of its appearance in the perikarya cells of trigeminal ganglia, decreasing the number of labelled neurons. A more marked inflammation led to the decline in the number of nervous cells containing HRP. The decreased number of labelled HRP cells was recorded during the regeneration phase. This difference in the dynamics of neuron labelling in trigeminal ganglia was associated with the damaging effect of high temperature and inflammation on HRP uptake and transportation.     

Roles of nitric oxide and prostaglandins in pathogenesis of delayed colonic transit after burn injury in rats

Burn injury has been shown to impair gut transit, but the exact mechanism remains unknown. The present study investigated whether nitric oxide synthase (NOS) and cyclooxygenase (COX) mediated changes in burn-induced colonic transit. After rats underwent 30% total body surface area burn injury, they were injected with S-methylisothiourea (SMT, selective inducible NOS inhibitor), 7-nitronidazole (7-NI, selective neuronal NOS inhibitor), and nimesulide (NIM, selective COX-2 inhibitor), respectively. The protein and mRNA of NOS and COX-2 were measured by Western blot analysis and real-time RT-RCR, and localization of NOS and COX-2 protein was determined by immunohistochemistry. Our results showed that colonic transit assessed by the geometric center was delayed from 3.47+/-0.28 in controls to 2.21+/-0.18 after burn (P<0.009). SMT and NIM significantly improved colonic transit in burned rats but had no effect in sham-operated rats. 7-NI failed to modify delayed transit in burned rats but significantly delayed colonic transit in sham-operated rats. Both protein and mRNA of inducible NOS and COX-2 increased significantly but not neuronal NOS in burned rats. Inducible NOS protein expression was noted not only in epithelial cells but also in neurons of the myenteric ganglia in burned rats. These findings suggest that nitric oxide (NO) produced by neuronal NOS plays an important role in mediating colonic transit under the physiological condition. NO produced by inducible NOS and prostaglandins synthesized by COX-2 are both involved in the pathogenesis of delayed colonic transit after burn injury. Inducible NOS expression in neurons of the myenteric ganglia may contribute to dysmotility with burn injury.     

Horseradish peroxidase localization of sympathetic postganglionic and parasympathetic preganglionic neurons innervating the monkey heart

The localization of the sympathetic postganglionic and parasympathetic preganglionic neurons innervating the monkey heart were investigated through retrograde axonal transport with horseradish peroxidase (HRP). HRP (4 mg or 30 mg) was injected into the subepicardial and myocardial layers in four different cardiac regions. The animals were euthanized 84-96 hours later and fixed by paraformaldehyde perfusion via the left ventricle. The brain stem and the paravertebral sympathetic ganglia from the superior cervical, middle cervical, and stellate ganglia down to the T9 ganglia were removed and processed for HRP identification. Following injection of HRP into the apex of the heart, the sinoatrial nodal region, or the right ventricle, HRP-labeled sympathetic neurons were found exclusively in the right superior cervical ganglion (64.8%) or in the left superior cervical ganglion (35%). Fewer labeled cells were found in the right stellate ganglia. After HRP injection into the left ventricle, labeled sympathetic cells were found chiefly in the left superior cervical ganglion (51%) or in the right superior cervical ganglion (38.6%); a few labeled cells were seen in the stellate ganglion bilaterally and in the left middle cervical ganglion. Also, in response to administration of HRP into the anterior part of the apex, anterior middle part of the right ventricle, posterior upper part of the left ventricle, or sinoatrial nodal region, HRP-labeled parasympathetic neurons were found in the nucleus ambiguus on both the right (74.8%) and left (25.2%) sides. No HRP-labeled cells were found in the dorsal motor nucleus of the vagus on either side.     

Non-synaptic interaction between neurons in molluscs

1. Isolated pedal ganglia of the pteropodial mollusc, Clione limacina, generate a locomotory rhythm. In 30% of the pedal ganglia preparations the locomotory rhythm was not regular, i.e. the locomotor generator worked in “bursts” alternating with periods of low activity.2. The “locomotor bursts” were caused by spontaneous activation of command neurons located in the pedal ganglia.3. A single neuron was extracted from burst-generating preparations by means of the intracellular microelectrode and then its soma was put back, into the initial place between the ganglion cells. Twenty-five percent of the isolated neurons renewed the bursts-related changes in their activity after the insertion into the ganglion. The neurons which were originally excited during the “locomotor bursts” continued to be excited after isolation, while those which were inhibited continued to be inhibited during the bursts.4. It is suggested that the command neurons controlling the locomotor generator can exert action on the target cells in the absence of morphological synapses.     

Quantitative Estimation of HRP-Labeled Sensory and Motor Neurons during Nerve-Dependent and Nerve-Independent Periods of Urodele Limb Regeneration

The relationship between urodele regeneration and the possibility of regeneration in mammals is unclear, but the idea of possible regeneration of neural elements in man is being studied because of its potential clinical importance. One of the great challenges is to gain sufficient knowledge about the basic biology of animal regeneration and to use it for the betterment of the mankind. It is known that the initial stages of urodele limb regeneration depend on the presence of intact nerve fibers connected to their cell bodies. The nerve fibers severed at the level of limb amputation regrow and penetrate the blastema, providing blastema cells with indispensable factors. These factors are produced in the perikarya of neurons and transported via their axons to the blastema. Numerous studies have been performed to elucidate the quantitative relationships between nerve fibers and limb regeneration. However, there are no reports dealing with the individual nerve cells at work. The aim of this investigation was to analyze the quantitative participation and qualitative distinctions of different nerve cells innervating the regenerating parts of the urodele limb and their possible roles in the nerve-dependent and nerve-independent periods of regeneration. The cells under study are housed in the dorsal ganglia (sensory neurons) and in the ventral part of the spinal cord gray matter (motor neurons). The direct involvement of these neurons in different regeneration periods was visualized by means of horseradish peroxidase (HRP) labeling. A total of 34 animals (21 experimental and 13 control) were used to study fluctuations in the numbers of labeled nerve cells. The results are summarized as follows: (a) the first nerve cells incorporating HRP within 5 days after amputation are found in the dorsal ganglia, whereas motor neurons in the gray matter are labeled within 7 days; (b) the number of labeled perikarya increases during the nerve-dependent regeneration period (0–21 days after amputation), with the percentage of implicated sensory neurons exceeding that found in the control series; and (c) during the next, nerve-independent period, the number of participating labeled neurons decreases gradually. Such fluctuations in the number of labeled neurons might represent the metabolic status of these cells in their effort to provide the blastema cells with the factors needed at the appropriate time. The current findings support previous observations that the periods of dependence and independence of urodele limb regeneration on the integrated control of brachial nerves reflect changes in the metabolism of individual sensory and motor neurons.     

Gastrin-cholecystokinin immunoreactivity in the central nervous system of Helix aspersa during rest and activity

The immunostaining pattern for the peptide gastrin/cholecystokinin 8 (gastrin/CCK8) in the molluscan central nervous system has been considered. The changes in the distribution of gastrin/CCK8 immunoreactivity were analyzed in the neurons of different areas of the cerebral ganglia (mesocerebrum and metacerebrum) and in the buccal ganglia of the terrestrial snail Helix aspersa, during rest and active phases. During the period of inactivity and after one day of activity, there were several immunoreactive neurons in the mesocerebrum and metacerebrum of the snails and in the buccal ganglia, whereas after 7 days of activity the number of labeled neurons decreased. Data suggested a storage of gastrin/CCK8 in the neurons when behavioral activities in which the peptide is involved (such as feeding-related behavior) are suppressed or reduced. The different percentage of gastrin/CCK8 immunoreactive neurons in the left and right mesocerebrum provides information about the activities controlled by these neurons, which could be related to the adaptive evolution and plasticity of the brain in terrestrial pulmonates.     

Effects of starvation on haemolymphatic glucose levels, glycogen contents and nucleotidase activities in different tissues of Helix aspersa (Müller, 1774) (Mollusca, Gastropoda)

In the present study, the glucose concentration in the haemolymph and glycogen levels were determined in the various body parts of the Helix aspersa snail after feeding lettuce ad libitum and after various periods of starvation. To characterize the effect of starvation on nucleotidase activity, enzyme assays were performed on membranes of the nervous ganglia and digestive gland. Results demonstrated the maintenance of the haemolymph glucose concentration for up to 30 days of starvation, probably due to the consumption of glycogen from the mantle. In the nervous ganglia, depletion of glycogen occurs progressively during the different periods of starvation. No significant changes were observed on ATP and ADP hydrolysis in the membranes of nervous ganglia and no alterations in Ca2+ -ATPase and Mg2+ -ATPase occurred in the membranes of the digestive gland of H. aspersa during the different periods of starvation. Although there were no changes in the enzyme activities during starvation, they could be modulated by effectors in situ with concomitant changes in products/reactants during starvation.     

The long-term fate of neurons in allografts of ganglia in AG-B-compatible normal and immunologically tolerant rats

Neurons in Ag-B-incompatible allografts of ganglia are acutely rejected while those in Ag-B-compatible grafts are able to survive the immune reaction directed against them. The present study was undertaken to determine the long-term fate of neurons in allografts of ganglia in Ag-B-compatible rats. Isogenic strains of Ag-B-compatible adult Lewis (LE) and Fischer (FR) rats were used. The sensory nodose ganglia were reciprocally exchanged between normal LE and FR and between LE and FR animals rendered immunologically tolerant of each other's histocompatibity antigens. The findings were similar in both rat strains and revealed that although neurons and glial cells (i.e., satellite and Schwann cells) could survive for prolonged periods they were nevertheless eventually rejected by normal (nonimmunosuppressed) recipients. On the other hand, neurons and glial cells survived indefinitely in allografts in tolerant rats. Moreover, these neurons were functional because they regenerated nerve fibers into cotransplanted isografts of tongue and exerted the neurotrophic influence of inducing taste bud regeneration. The results demonstrate that, unlike kidney and heart, neurons in ganglia allografts cannot survive indefinitely without immunosuppression in Ag-B-compatible animals. Nevertheless, the permanent survival and function of neurons in Ag-B-compatible grafts can be achieved, as it is in Ag-B-incompatible ganglia grafts, by rendering the recipient immunologically tolerant.     

Metabolic changes in the basal ganglia of patients with Huntington's disease: an in situ hybridization study of cytochrome oxidase subunit I mRNA

On the basis of the functional model of the basal ganglia developed in the 1980s and the neuropathological findings in Huntington's disease (HD), changes in the neuronal activity of the basal ganglia have previously been proposed to explain the abnormal movements observed in this pathology. In particular, it has been stated that the neurodegenerative process affecting the basal ganglia in the disease should provoke a hypoactivity in the internal segment of the pallidum (GPi) that could explain choreic movements observed in the disease. To test this functional hypothesis, we performed an in situ hybridization study on control and HD brains postmortem, taking cytochrome oxidase subunit I (COI) mRNAs expression as index of neuronal activity. As most of the HD patients studied were under chronic neuroleptic (NL) treatment, we also studied the brains of non-HD patients under chronic NL treatment. Our results show that in HD brain the number of neurons expressing COI mRNA tends to be lower in the striatum, GPe and GPi, suggesting a severe involvement of these structures during the neurodegenerative process. Moreover, COI mRNA level of expression was markedly reduced within neurons of the putamen and GPe. Surprisingly, COI mRNA expression was not modified in the GPi in HD brains compared with controls. This paradoxical result in the GPi may be explained by the antagonistic effect of GPe hypoactivity and the degenerative process involving neurons of GPi. Our results indicate that the functional modifications, and consequently the pathophysiology of abnormal movements, observed in HD basal ganglia are more complex than expected from the currently accepted model of the basal ganglia organization.     

Cytoplasmic type 80S ribosomes associated with yeast mitochondria. IV. Attachment of ribosomes to the outer membrane of isolated mitochondria

Cultures of whole fetal rat sensory ganglia which had matured and myelinated in culture were treated for 1-3 h with a pulse of 0.2% trypsin. The tissue was observed during the period of treatment and during subsequent weeks using both light and electron microscopy. Within minutes after trypsin addition the matrix of the culture was altered and the nerve fascicles loosened. Progressive changes included the retraction of Schwann cell processes from the nodal region the detachment of the myelin-related paranodal Schwann cell loops from the axon, and lengthening of the nodal region as the axon was bared. The retraction of myelin from nodal stabilized several hours after trypsin withdrawal. Breakdown of the altered myelin segments was rare. There were no discernable changes in neurons or their processes after this exposure to trypsin. The partial repair which occured over a period of several weeks included the reattachment of paranodal Schwann cell loops to the axolemma and the insertion of new myelin segments where a substantial length of axolemma had been bared. The significance of these observations to the characterization of the Schwann cell-axolemmal junctions on myelinated nerve fibers is discussed. The dramatic degree of myelin change that can occur without concomitant myelin breakdown is particularly noted, as is the observation that these altered myelin segments are, in part, repaired.     

Long-term transplantation of mature sensory neurons

The spinal ganglia were transplanted into the mesocolon of adult cats for periods of from 1 day to 9 months. About 30% of differentiated sensory neurons survived during the longterm transplantation. The intensive regeneration of the sensory neurons processes was characteristic of the transplanted neurons. Total myelinization of the regenerating nerves occurred during the 3rd--5th month. Potential regeneration capacity of the differentiated neurons and possibly of their prolonged transplantation were revealed.     

Nerve growth factor-independent development of embryonic mouse sympathetic neurons in dissociated cell culture

Although ganglia from neonatal mouse sympathetic ganglia require nerve growth factor (NGF) for survival in culture, explanted sympathetic ganglia from early embryonic stages do not require added NGF for survival and growth. To determine whether the change in growth factor requirement is due to changes in the neurons themselves, to variations in neuronal populations, or to changes in nonneuronal cells, we examined the response to growth factors by dissociated sympathetic neurons at various stages of development. Results indicate that neurons from the 14-day gestational (E14) superior cervical ganglion (SCG) do not require NGF for initial survival and neurite extension, but do require the conditioned medium neurite extension factor, CMF. By 2 to 3 days thereafter, whether in vivo or in culture, most neurons have developed a requirement for NGF for survival in culture. During the same period, there is a concomitant increase in responsiveness to NGF alone as a trophic agent. Changes in response to NGF are not due to changes in NGF content of ganglia, to interactions in culture with nonneuronal cells, or to age-related differences in NGF requirements for maximum survival. The changes in growth factor requirements may be related to mechanisms regulating specificity of nerve-target connections.     

Differences in neuronal lipid composition between superior cervical ganglia and nodose ganglia of the rat

The lipid content and composition of rat superior cervical ganglia containing sympathetic motor neurons and nodose ganglia containing parasympathetic sensory neurons were studied for the first time to elucidate the mechanism of the different effects of exogenous gangliosides on these neurons in the culture medium. The ganglioside content of the superior cervical ganglia was almost 3-times that of the nodose ganglia. Although both ganglia contained GM3, GD3, GD1b and GT1b as major gangliosides, the nodose ganglia additionally contained a significant amount of sialosyllactoneotetraosylceramide LM1 (10% of total sialic acids). Contrasting with nodose ganglia, vagus fiber and dorsal root ganglia of rats, superior cervical ganglia had a higher content of sulfatide than galactosylceramide. The phospholipid content was lower in superior cervical ganglia than in nodose ganglia. Superior cervical ganglia contained less ethanolamine plasmalogen and more phosphatidylcholine than nodose ganglia. Sphingomyelin in superior cervical ganglia contained mainly medium-chain fatty acids, while that in nodose ganglia contained mainly longer-chain fatty acids. Differences in the fatty acid composition of glycerophospholipids were also observed. The results indicate that the properties of neuronal cell membranes from superior cervical ganglia and nodose ganglia are quite different, and that the differences may reflect the physiological roles of these ganglia.     

Temporal Changes of CB1 Cannabinoid Receptor in the Basal Ganglia as a Possible Structure-Specific Plasticity Process in 6-OHDA Lesioned Rats

The endocannabinoid system has been implicated in several neurobiological processes, including neurodegeneration, neuroprotection and neuronal plasticity. The CB1 cannabinoid receptors are abundantly expressed in the basal ganglia, the circuitry that is mostly affected in Parkinson’s Disease (PD). Some studies show variation of CB1 expression in basal ganglia in different animal models of PD, however the results are quite controversial, due to the differences in the procedures employed to induce the parkinsonism and the periods analyzed after the lesion. The present study evaluated the CB1 expression in four basal ganglia structures, namely striatum, external globus pallidus (EGP), internal globus pallidus (IGP) and substantia nigra pars reticulata (SNpr) of rats 1, 5, 10, 20, and 60 days after unilateral intrastriatal 6-hydroxydopamine injections, that causes retrograde dopaminergic degeneration. We also investigated tyrosine hydroxylase (TH), parvalbumin, calbindin and glutamic acid decarboxylase (GAD) expression to verify the status of dopaminergic and GABAergic systems. We observed a structure-specific modulation of CB1 expression at different periods after lesions. In general, there were no changes in the striatum, decreased CB1 in IGP and SNpr and increased CB1 in EGP, but this increase was not sustained over time. No changes in GAD and parvalbumin expression were observed in basal ganglia, whereas TH levels were decreased and the calbindin increased in striatum in short periods after lesion. We believe that the structure-specific variation of CB1 in basal ganglia in the 6-hydroxydopamine PD model could be related to a compensatory process involving the GABAergic transmission, which is impaired due to the lack of dopamine. Our data, therefore, suggest that the changes of CB1 and calbindin expression may represent a plasticity process in this PD model.