Retrograde axonal transport of bismuth: an autometallographic study.

Bismuth subnitrate was injected into the triceps surae muscle of 3-month-old male Wistar rats. Sections of lumbar spinal cord (L4-L6) and corresponding dorsal root ganglia were developed by autometallography (AMG) to trace possible bismuth in neuronal somata resulting from retrograde axonal transport. At 3 days after treatment bismuth clusters could be traced by AMG in spinal cord motor neurons and in dorsal root ganglion cells ipsilateral to the injection site. Retrograde transport of bismuth could be avoided by ligation or intraneuronal injection of cholchicine into the sciatic nerve. At the ultrastructural level bismuth was found to be located exclusively in lysosome-like organelles in motor and sensory neuronal somata projecting to the injection site. The present study shows that bismuth is transported retrogradely in both sensory and motor axons if their terminals are exposed to bismuth ions.

Neuron loss in cerebellar cortex of rats exposed to mercury vapor: a stereological study.

Mercury vapor produces tremor in humans and experimental animals. We have previously reported that mercury vapor intoxication over an 8-week period induces only subtle changes in dorsal root ganglia and nerve roots in rats. In the present study we have carried out stereological analyses of the cerebellum of the same rats, and demonstrated significant losses of Purkinje cells (12.7%, 2P = 0.005) and granule cells (15.6%, 2P = 0.016). All sizes of Purkinje cells were lost with an equal probability, i.e. there were no indication of any preferential loss of any subpopulation of the neurons. The volume of the granular cell layer was significantly reduced (18.9%, 2P = 0.0 15), whereas the volumes of the molecular layer and the white matter were unchanged. Previous stereological studies have demonstrated that methyl mercury intoxication primarily induces degeneration in the peripheral nervous system, while sparing the cerebellum. We therefore suggest that metallic mercury vapor and methyl mercury have different toxicological profiles in rats, where metallic mercury vapor mainly affects the central nervous system and methyl mercury mainly affects the peripheral nervous system.

Autometallographic mercury correlates with degenerative changes in dorsal root ganglia of rats intoxicated with organic mercury.

Organic mercury intoxication in rats produces degenerative changes in the dorsal root ganglia and dorsal nerve roots. In a previous study of rats treated with organic mercury (2 mg/kg ) for 19 days, significant losses of ganglion cells (especially A-cells) and myelinated axons were observed in dorsal nerve roots and there was qualitative evidence of glial cell proliferation and the formation of Nageotte bodies (1). In the present study, the autometallographic silver-enhancement technique, for tracing inorganic mercury bound to sulphide or selenide (AMG-Hg), was applied to tissue sections of dorsal root ganglia and dorsal nerve roots of the same rats used in the earlier study. Satellite cells and macrophages that surrounded ganglion cells and formed Nageotte bodies were heavily labelled by coarse deposits of AMG-Hg, while the labelling of ganglion cells was less pronounced. A-cells were consistently labelled, while B-cells were only occasionally labelled. In the dorsal nerve roots, only a few AMG-Hg deposits could be seen in macrophages. At the ultrastructural level, AMG-Hg was observed within lysosomes of target cells. It is concluded that AMG-Hg is primarily located in glial cells and that the pattern of deposition of AMG-Hg is the same as that for the morphological changes observed in rats intoxicated with organic mercury.

A stereological study of dorsal root ganglion cells and nerve root fibers from rats exposed to mercury vapor.

Although mercury vapor is known to produce tremor and peripheral neuropathy, neuropathological studies of the effects of the vapor are few in number. The aim of the present study has been to evaluate the effect of mercury vapor on the morphology of the dorsal root ganglion and the spinal nerve roots. Adult male rats were exposed to mercury vapor for 33 days. The exposed rats developed somatic signs of intoxication and became increasingly irritable. The total numbers and volumes of A- and B-cell perikarya in the dorsal root ganglia, the total number of myelinated axons in the roots, and the cross-sectional areas of axon and myelin in the nerve roots were estimated using unbiased stereological principles. The mean cross-sectional area of myelin associated with nerve fibers in dorsal nerve roots of the exposed group was significantly reduced by 20% (2P=0.014). A tendency towards a reduction was seen in axon area of myelinated nerve fibers in the dorsal nerve roots (2P=0.087) and in the total numbers and mean volume of A-cell perikarya (2P = 0.059 and 2P=0.087, respectively). No differences between the two test groups were found for any of the parameters measured in B-cells and ventral nerve roots. It is concluded that mercury vapor, in a dose sufficient to produce intoxication, induces only minor changes in dorsal root ganglion and nerve roots in rats.

Selective degeneration of dorsal root ganglia and dorsal nerve roots in methyl mercury-intoxicated rats: a stereological study.

The components of the nervous system of rats that are most critically affected by methyl mercury are still a matter of debate. A recent stereological study of rats with typical symptoms resulting from methyl mercury intoxication demonstrated that the morphology of cerebellar granule cells and Purkinje cells were unchanged at the light microscopic level, even though there was pronounced degeneration of myelinated axons in dorsal nerve root nerves. In the present study, unbiased stereological methods were used to quantify morphological changes in the dorsal root ganglion, and dorsal and ventral nerve roots of the rats used in the previous study. The rats were treated with methyl mercury (2 mg daily/kg, per os) for a 19-day period that was followed by a 32-day period without treatment. The means of the total numbers of A-cell and B-cell perikarya in the dorsal root ganglion of the intoxicated rats were reduced by 60% and 24%, respectively. The mean volume of A-cell perikarya in rats of the experimental group was reduced by 22%, whereas the mean volume of B-cell perikarya was the same in the two groups. In the experimental group, the total number of myelinated axons in the dorsal nerve roots was reduced by 60%, whereas no difference was found in the ventral nerve roots. The areas of axon and myelin sheath, dorsal and ventral nerve roots were not affected. This study demonstrates that extensive loss of dorsal root ganglion cells and myelinated axons in dorsal nerve roots precedes light microscopical changes in the ventral nerve roots and the cerebellum of rats intoxicated with methyl mercury.

[Arrhythmogenic right ventricular cardiomyopathy]. / Arytmogen højre ventrikel kardiomyopati.

Arrhythmogenic right ventricular dysplasia is a rare cardiomyopathy, but a frequent cause of ventricular tachyarrhythmia and sudden cardiac death among young otherwise healthy individuals. This article contains a review of the current knowledge on epidemiology, diagnosis, symptoms and signs as well as theories on etiology and pathogenesis, prognosis and treatment. The aim is to draw attention to the disease as a cause of syncope, ventricular tachycardia and sudden cardiac death.

A stereological study of dorsal root ganglion cells and nerve root fibers from rats treated with inorganic mercury.

Unbiased stereological methods have been used to quantify the effects of inorganic mercury on the morphology of the fifth lumbar dorsal root ganglion cells and nerve root fibers. Adult male Wistar rats were treated with intraperitoneal injections of mercuric chloride (0.15 mg daily) for 30 days. The total numbers and mean volumes of A- and B-cell perikarya were estimated using the optical fractionator and the vertical rotator techniques. The total numbers of myelinated axons in the ventral and the dorsal roots were estimated with the two-dimensional fractionator technique and the areas of axon and myelin were estimated using the two-dimensional nucleator technique. No differences were found for any parameters in experimental and control animals, indicating that inorganic mercury intoxication alters neither the numbers or sizes of dorsal root ganglion cells and nerve root fibers nor the amount of myelin associated with the nerve fibers.

The effect of selenium on the localization of autometallographic mercury in dorsal root ganglia of rats.

The autometallographic technique was used to demonstrate the localization of mercury in dorsal root ganglia of adult Wistar rats. The animals were either exposed to mercury vapour, 100 micrograms Hg m-3, 6 h day-1, 5 days per week, or treated with organic mercury in the drinking water, 20 mg CH3HgCl per litre, for 4 weeks. The effect of orally administered sodium selenite on the pattern of intracellular distribution of mercury in these two situations was investigated. In rats exposed to mercury vapour alone, faint staining was present in ganglion cells. The selenite induced a conspicuous increase in the number of stained cells and in the intracellular staining intensity. In rats treated with organic mercury, mercury deposits were detected within ganglion cells and macrophages. The number of mercury-containing cells was increased by co-administration of selenite. In addition, satellite cells, the capsule and vessel walls were faintly stained. Twenty weeks after cessation of the organic mercury treatment, mercury staining was reduced. Again, selenite treatment enhanced staining intensity. When studied using the electron microscope, mercury was restricted to lysosomes, irrespective of treatments. The present study shows that the deposition of autometallographic mercury in the dorsal root ganglia depends on the chemical type of mercury, the co-administration of selenite and the length of the survival period.

Arrhythmogenic right ventricular dysplasia as a cause of sudden death.

Arrhythmogenic right ventricular dysplasia (ARVD) is a poorly understood and often underdiagnosed disorder of the right ventricle, characterized by replacement of myocardium by fibroadipose tissue, arrhythmic manifestations, and sudden death. The disease occurs in families and is inherited as an autosomal dominant trait. This report describes five cases of ARVD identified by autopsy. In three of the cases, sudden death occurred in the young (16-28 years old) during or shortly after exercise. In another case, a 46-year-old man with no relevant medical history was found dead in his bathroom. In the last case, a 57-year-old woman died from pulmonary thromboembolism. In none of the subjects had the disease been diagnosed or suspected before death. Only one (a 21-year-old man) had previous typical symptoms of the disease. Autopsy examination showed right ventricle dilation and, in four cases, cardiomegaly. The right ventricular myocardium of all hearts was almost replaced by adipose tissue and to a variable degree by fibrous tissue, while the left ventricle myocardium demonstrated no, or only scattered, fibro-fatty infiltration. Postmortem diagnosis of ARVD can be important in identifying possible affected family members in order to initiate treatment.

Mercury content in rat teeth after administration of organic and inorganic mercury. The effects of interrupted exposure and of selenite.

Rat molars are indicators of exposure concentration and target organ content in chronic mercury vapor exposure. We wished to study the accumulation and persistence of organic and inorganic mercury in rat teeth and the effect of selenium on mercury retention. Male Wistar rats received either inorganic or organic mercury (with or without addition of selenite), selenite only, or no mercury or selenite (controls) in the drinking water for 4 weeks. Group A was killed after exposure. Group B was killed 20 weeks later. The mercury content was measured by cold-vapor atomic absorption spectrophotometry. The mercury content in the molars in group B was 66% and 77% less than in group A after inorganic and organic exposure, respectively. In the incisors the corresponding reductions were 90% and 97%. Selenite had limited effect on mercury retention in group A and none in group B. We suggest that rat molars and, by inference, human deciduous teeth may serve as indicators of organic and inorganic mercury exposure.

Autometallographic demonstration of mercury in rat molars.

Male, adult Wistar rats were exposed to 500 micrograms/m3 mercury vapor 6 h per day, 5 days a week for 4 wk. They were subsequently killed by transcardial perfusion. The molars were extracted, demineralized, and embedded in resin before sectioning. Autometallographic development was performed according to the method of Danscher & Möller-Madsen. Mercury deposits were found in small amounts in several areas of the pulp, but with larger accumulations of grains in relation to odontoblasts. Mercury also could be seen in odontoblastic processes in the dentin and predentin. Our conclusion is therefore that systemic uptake of mercury vapor leads to accumulation of mercury in the odontoblasts and that the mercury may be transported into the dentin tubules in the odontoblastic process.

Retrograde axonal transport of mercury in rat sciatic nerve.

Mercuric chloride was injected into the triceps surae muscle of adult male Wistar rats. Following a survival period of 1, 2, 3, 6, or 100 days, the rats were euthanized by transcardial perfusion. Sections of lumbar spinal cord segments (L4-L6) with corresponding dorsal root ganglia were subjected to the autometallographical silver-enhancement technique in order to localize mercury at both light and electron microscopical levels. Intramuscular injections of mercury resulted in ipsilateral accumulations of mercury in ventral horn motoneurons and dorsal root ganglion cells after a survival period of 2 days. Mercury deposits were still present when animals were allowed to survive 100 days. The mercury staining was suppressed ipsilateral to the intramuscular mercury injection by a microinjection of colchicine into the sciatic nerve or by ligation of the sciatic nerve. At the ultrastructural level, mercury was detected within lysosomes of target cells. The findings indicate that mercury is transported retrogradely in axons of ventral horn motoneurons and dorsal root ganglion cells.

Detection of mercury in rat spinal cord and dorsal root ganglia after exposure to mercury vapor.

Adult male Wistar rats were exposed to mercury vapor, 50 micrograms Hg/m3, 6 hr/day, 5 days/week, over 1-, 2-, 3-, 4-, 6-, and 8-week periods. Sections from the spinal cord and dorsal root ganglia from spinal levels C1, C5, T6, and L1 were stained with the autometallographical technique and the distribution of mercury deposits described at light and electron microscopical levels. A quantitative analysis of the amount of mercury in blocks of the spinal cord was performed using cold vapor atomic absorption spectrophotometry. After an exposure period of 2 weeks, silver-enhanced mercury grains could be observed in spinal cord neurons located in Rexed laminae IV-X. Ventral horn motoneurons were heavily stained in all of the spinal cord segments. Ependymal cells and glial cells of both the spinal gray and white matter contained cytoplasmatic mercury accumulations in rats exposed to mercury vapor for 4 weeks. In the dorsal root ganglia, only ganglion cells showed a faint mercury staining and the amount of staining was notably less than that seen in the ventral horn motoneurons. At the ultrastructural level, mercury was seen primarily within lysosomes of target cells. The quantitative mercury measurements demonstrated that spinal cords from rats exposed to mercury vapor for 6 or 8 weeks contained a significantly higher concentration of mercury than those from control animals.

Differentiation of silver-enhanced mercury and gold in tissue sections of rat dorsal root ganglia.

Autometallography was used in conjunction with light and electron microscopy to detect traces of gold and mercury in the dorsal root ganglia of rats treated with sodium aurothiomalate and mercuric chloride. In order to differentiate between gold and mercury in tissue sections, the gold accumulations were removed by potassium cyanide, leaving mercury sulphides/selenides as the only possible catalysts for autometallographic development. With this technique, it is now possible to differentiate between all tissue metals capable of initiating the autometallographic process, i.e. gold, vesicular zinc, and sulphides and selenides of mercury and silver.

Autometallographic detection of gold in dorsal root ganglia of rats treated with sodium aurothiomalate.

Ultraviolet light autometallography, a very sensitive method for gold detection, was applied to sections of dorsal root ganglia from adult male Wistar rats treated with intraperitoneal injections of sodium aurothiomalate. Silver-amplified traces of gold were detected within the cytoplasm of ganglion cells, satellite cells, Schwann cells, macrophages, endothelial cells, and fibroblasts throughout the ganglia. Gold was never detected in axons nor myelin sheaths. In the electron microscope, gold deposits were restricted to the lysosomes irrespective of cell type or dosage.

Autometallographic silver-enhancement of colloidal gold particles used to label phagocytic cells.

The present paper demonstrates that colloidal gold silver-enhanced by autometallography (AMG) can be used to label phagocytic cells for light microscopic detection. Cultured macrophages were exposed to 0.5 microliters 6 nm colloidal gold particles for 24 or 48 h. Other cultures were exposed to 25 microliters of the same solution for 1 to 14 days. The staining was found to be stable also when new unmarked cells were applied. The colloidal gold had no adverse effect on the cells. The presented technique might also prove valuable for estimation of the total number of phagocytes in a culture or in an organism by applying labelled cells to culture or organism, and to ascertain the fate of a population of marked cells.

Autometallographic detection of mercury in rat spinal cord after treatment with organic mercury.

Autometallography was used to localize mercury in rat spinal cord after intraperitoneal administration of methylmercuric chloride (200 micrograms CH3HgCl daily). The technique permits small amounts of mercury sulfides and mercury selenides to be visualized by silver-enhancement. Mercury deposits were observed by light microscopy only in neurons. In all of the spinal cord segments selected (first cervical segment, C1; fifth cervical segment, C5; sixth thoracic segment, T6; and first lumbar segment, L1) the mercury was observed with cumulative dosages of 6000 micrograms CH3HgCl and greater. Laminae VII, VIII, and IX contained the majority of stained neurons, whereas laminae IV, V, VI, and X had a relatively lower density of mercury-containing neurons. Stained neurons were confined to specific cell groups, such as Clarke's column, nucleus intermedio-lateralis, nucleus cervicalis centralis, and nucleus dorsomedialis. At the ultrastructural level, mercury deposits were restricted to lysosomes of neurons and occasional accumulations in the lysosomes of ependymal cells.

Mercury in the dorsal root ganglia of rats treated with inorganic or organic mercury.

Autometallographic silver amplification has been used to demonstrate the localization of mercury deposits in rat dorsal root ganglia after repeated intraperitoneal injections of mercuric chloride or methylmercuric chloride. The silver-enhanced mercury deposits were demonstrated with the light and electron microscope. The degree of intracellular staining of the individual cells depended on the mercury compound and total dosage. Ganglion cells (types A and B) and macrophages were found to accumulate mercury after a total dosage of 400 micrograms HgCl2. After 600 micrograms HgCl2, satellite cells, endothelial cells and fibroblasts were additionally found to contain mercury deposits. Treatment with 6000 micrograms CH3HgCl caused faint staining of type A and B ganglion cells and fibroblasts. Macrophages, however, were the most heavily stained cells after treatment with CH3HgCl. Ultrastructurally, mercury was exclusively located in lysosomes. This was irrespective of the cell type and mercury compound used for treatment.