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Biomedical subjects

G Danscher

Publications and source records attributed to G Danscher.

At least 73 records · Page 4Linked to original sources

How to detect gold, silver and mercury in human brain and other tissues by autometallographic silver amplification.

Gold, silver, mercury and zinc bind chemically to sulphide or selenide ions and create crystal lattices that can be detected in histological sections by a silver amplification technique called autometallography (AMG). The technique specifically magnifies such nanometer-sized catalytic crystals. For each metal, a detailed protocol has been worked out. If several different AMG metals/metal molecules are present in the same tissue, it is possible to distinguish one from another. The AMG technique is based on the capability of small crystal lattices of the aforementioned metals and metal molecules to initiate AMG silver amplification. Electrons released from adhering hydroquinone molecules reduce silver ions that are integrally connected with the crystal lattices. In this manner, particles consisting of only a few atoms of, say, gold, or molecules of mercury selenide (Figure 1), can be silver amplified to a size at which they can be detected in the electron microscope, or even further to dimensions that can be observed in the light microscope. Thus the AMG technique opens up the possibility of visualizing gold, e.g. in the nervous system of rheumatic patients who have been treated with aurothiomalate. Mercury can similarly be visualized in tissues from individuals who have been exposed to mercury, either through leaching from amalgam dental fillings, through eating fish, or by occupational exposure, and silver in the central (CNS) and peripheral nervous systems (PNS) and other tissues from individuals exposed to silver in one form or another. In the future, the possibility of demonstrating vesicular zinc, a particular pool of endogenous zinc that is found in terminals of zinc-enriched neurons (ZEN neurons), might prove valuable for pathological interpretation of diseases such as Alzheimer's disease. The vesicular zinc, present in some of the synaptic vesicles of ZEN neuron terminals, is most impressively demonstrated by AMG in telencephalic structures. It is becoming increasingly indisputable that vesicular zinc is related to synaptic activity influencing or modulating facilitatory synapses. ZEN neurons are probably a sub-population of glutaminergic neurons. A technique for the post-mortem demonstration of vesicular zinc in terminals of ZEN neurons in human brains is therefore urgently required.

Animals↗

The von Kossa reaction for calcium deposits: silver lactate staining increases sensitivity and reduces background.

The classical von Kossa method has been modified: the high silver nitrate concentration in the original was replaced by 0.05% silver lactate with hydroquinone remaining the reducing agent of choice. The present modification stained calcification nodules with a sensitivity comparable to the original von Kossa reaction, but resulted in a reduced background staining in cultured osteoblasts. The method works well also with plastic- or paraffin-embedded tissue sections.

Calcification, Physiologic↗

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.

Animals↗

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.

Animals↗

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.

Animals↗

Retrograde tracing of zinc-containing neurons by selenide ions: a survey of seven selenium compounds.

The autometallographic retrograde tracing of zinc-containing neurons by intracerebral injection of sodium selenite (Na2SeO3), introduced by Danscher in 1982, has recently been described in more detail. Intracerebral injections of both sodium selenide (Na2Se) and sodium selenite (Na2SeO3) have been successfully used; however, sodium selenite had a rather toxic effect on the injected tissue. In the present study, we tested seven different selenium compounds to find the most suitable compound for retrograde tracing of zinc-positive pathways. Among the tested compounds, sodium selenide (Na2Se) caused insignificant necrosis within the injection site and was easily transported retrogradely when handled anaerobically. Sodium selenide is therefore recommended as the compound of choice.

Animals↗

Densitometric analysis of the local bleaching of the Neo-Timm staining pattern following intrahippocampal injection of diethyldithiocarbamate.

Treatment with certain metal chelating agents causes a time-dependent bleaching of the Neo-Timm staining pattern of zinc visualized in synaptic vesicles. In the present study, the extent and time course of the reversible chelation of hippocampal vesicular zinc was investigated following intrahippocampal injection of the chelating agent diethyldithiocarbamate. The carbamate (1.0 microliters 45 mg ml-1, 200 mM) was injected unilaterally into the hippocampal region of adult rats, which were allowed to survive 15 min-6 h before sacrifice. Control animals either received injections of distilled water or were untreated. Computerized optical densitometry was performed on cryostat sections of brains stained with the Neo-Timm method. Injection of diethyldithiocarbamate into the hippocampal region resulted in a localized bleaching of the Neo-Timm staining pattern. The extent of the bleaching varied with time being most pronounced at 15 min survival and gradually decreasing with time. After 6 h survival, a faint bleaching of the injected hippocampal region was barely seen. Computerized optical densitometry confirmed and extended the observations providing a semi-quantitative measure of zinc in synaptic vesicles.

Animals↗

Localization of mercury in CNS of the rat. IV. The effect of selenium on orally administered organic and inorganic mercury.

The distribution and exact cellular localization of mercury in the brain and upper cervical spinal cord of the adult male Wistar rat has been determined using the autometallographic silver-enhancement technique. A detailed atlas of mercury-containing nuclei following oral administration of HgCl2 (20 mg x liter-1 or CH3HgCl (20 mg x liter-1) was prepared. The effect of orally administered Na2SeO3 (2 mg x liter-1) on these patterns was investigated. In animals treated with CH3HgCl, sodium selenite induced a conspicuous increase in mercury staining of nerve cell bodies in specific areas of the central nervous system (CNS) including laminae III-VI in the cerebral cortex, thalamus, hypothalamus, and brain stem nuclei. In the cerebellum, the cortical Purkinje cells and nerve cells in the deep nuclei were targets for appreciable mercury accumulations after CH3HgCl. Again, these deposits were increased by coadministration of selenite. In the spinal cord following administration of CH3HgCl alone, staining was limited to the gray matter. The intensity of this staining was increased by selenite and deposits also appeared in the white matter. Mercury accumulations were present in scattered glia cells in the cuneate and gracile fasciculi. Treatment with HgCl2 alone or in combination with selenite yielded no staining of the Purkinje cells, nor did selenite result in an increase in the density of other stained cell bodies throughout the CNS, as was the case with organic mercury. The most intense neuronal staining was seen in sections taken from rats treated with a combination of CH3HgCl and selenite. Lesser staining was seen in neuroglia, ependymal, and choroidal cells. In the latter two cell types, staining intensity was unaffected by selenite treatment. In HgCl2-treated rats the same cell types were targets for mercury deposits although staining was to a significantly lesser degree. Concurrent treatment with selenite had no visible effect on the staining pattern. Ultrastructurally, the bulk of the mercury was located in lysosomes. Administration of CH3HgCl combined with selenite caused mercury to appear in the nuclei of neurons. Selenium treatment delayed the functional toxic effects of CH3HgCl. Sections prepared from animals treated separately with selenium or demineralized water (used as the solvent for all compounds) were devoid of mercury deposits.

Animals↗

Ultrastructural demonstration of mercury in Sertoli and Leydig cells of the rat following methyl mercuric chloride or mercuric chloride treatment.

The autometallographic silver enhancement technique has been used to demonstrate the ultrastructural localization of mercury in the testes of adult rats. Administration of mercuric chloride or methyl mercuric chloride in the drinking water (20 mg/L for 12 weeks) resulted in intracellular accumulations of mercury in the interstitial Leydig cells as well as in the Sertoli cells of the seminiferous tubules.

Animals↗

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.

Animals↗

Applications of autometallography to heavy metal toxicology.

Application of autometallography (AMG) to histological material from humans and animals exposed to gold, silver and mercury has made it possible to localize these heavy metals at light and electron microscopic levels. Because of high sensitivity of the technique, traces of the three metals have been demonstrated in tissues and cells that had previously not been suspected of containing metals. A chelatable pool of zinc in the synaptic vesicles of the zinc-positive neurones can be demonstrated by AMG in the brain. The well defined staining pattern can be used to estimate volumes of cortical subdivisions. Volumetric studies based on autometallographic differentiation of cortical regions have provided valuable information about the effects of different toxicants. AMG can be combined with new quantitative methods, such as electron energy loss spectroscopy (EELS), electron probe X-ray microanalysis (EPMA) and laser microprobe mass analysis (LAMMA), to enhance detection of AMG metal catalysts with these techniques.

Animals↗