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At least 19 recordsLinked to original sources
PubMed1978

Selective inhibition of potassium contracture in guinea pig taenia coli by ruthenium red.

Effects of ruthenium red on isotonic KCl induced contracture (K-contracture), cellular 45Ca uptake and 45 Ca binding to surface membranes were examined in the smooth muscle cells of guinea pig taenia coli. These results were compared with those using lanthanum (La3+). The tonic component of the K-contracture was selectively inhibited by 1 mM ruthenium red. In contrast, 1 mM La3+ inhibited the phasic component of the K-contracture to a large extent. Use of 1 mM ruthenium red selectively inhibited the tonic component of K-contracture and caused a marked decrease in cellular 45Ca uptake in that component of K-contracture. In contrast, 1 mM La3+ largely inhibited the phasic component and caused a significant decrease in cellular 45Ca in that component. According to Scatchard plot analysis, there are two kinds of Ca2+ binding sites of high and low affinity, respectively, on the surface membrane of the taenia coli. One mM ruthenium red suppressed those of low affinity more strongly than those of high affinity. In contrast, 1 mM La3+ suppressed high affinity sites more markedly than low affinity sites. Based on these results, it seems possible to conclude that ruthenium red mainly blocks the initial binding sites linked with Ca2+ influx which is related to the production of the tonic component while La3+ blocks those sites related to the phasic component of the K-contracture of guinea pig taenia coli.

Animals↗
PubMed1979

Potentiation and inhibition of ganglionic transmission by ruthenium red.

The effect of ruthenium red, 2.5 to 5 muM, on ganglionic transmission in rat superior cervical ganglia and frog abdominal ganglia were studied in vitro. In rat ganglia, ruthenium red caused a spontaneous firing of ganglia cells, and an increase in the amplitude and duration of the compound action potential following a single stimulus volley. However, transmission following a conditioning volley or a repetitive stimulus train to the preganglionic nerve was depressed up to 60 sec. The asynchronous firing caused by bethanechol was potentiated by ruthenium red. In the frog, ruthenium red caused repetitive firing of ganglion neurons following either orthodromic or antidromic stimulation. It is suggested that the potentiation of the single potential and the spontaneous firing are due to a ruthenium red-induced increase in intracellular calcium concentration. The depression of transmission may be due to a temporary depletion of readily releasable acetylcholine. It is also suggested that ruthenium red has an effect on the postsynaptic membrane.

Action Potentials↗
PubMed1976

Muscle satellite cells in urodele amphibians: faciliatated identification of satellite cells using ruthenium red staining.

The ruthenium red (RR) stained forelimb musculature of three species of urodeles Triturus (Notophthalmus) viridescens, Amblystoma maculatum, Amblystoma opacum in various stages of growth were examined with the electron microscope for the presence of satellite cells. It was found that RR staining facilitated greatly the identification of satellite cells. In young larvae of all three species satellite cells were detected with a frequency of 29% to 48% per total number of nuclei. In adult Triturus and Amblystoma maculatum satellite cells were no longer detected; instead "pericytes" as described by Hay ('74) were seen with a frequency of 12% and 3% respectively. During metamorphosis of Triturus satellite cells, with part of their myofiber-satellite cell intercellular space filled with basement membrane material, occurred at a peak frequency. The cells presumably are intermediate in the formation of "pericytes." At ten days after metamorphosis satellite cells and intermediate cells were no longer detected and the limb musculature contained only "pericytes" similar to the ones observed in adult newts. The significance of the presence of satellite cells in relation to limb regeneration and muscle regeneration is discussed.

Ambystoma↗
PubMed1976

Convulsions or flaccid paralysis induced by ruthenium red depending on route of administration.

Ruthenium red was administered to mice and cats intracranially or intraperitoneally. In mice, intracisternal administration produced status epilepticus and tonic convulsions. In contrast, intraperitoneal administration induced total flaccid paralysis lasting several hours. These effects of Ruthenium red were partially blocked by the simultaneous administration of CaCl2. EDTA, at doses much greater than those of Ruthenium red, produced effects similar to those of the dye, which were also blocked by CaCl2 administration. In cats, intraventricular or intrahippocampal administration of Ruthenium red through a permanently implanted cannula produced after a few minutes subclinical paroxysmal activity in all brain regions recorded. After several hours the animals developed typical grand mal seizures. Intraperitoneal injection of Ruthenium red to cats did not affect the EEG but markedly depressed muscular activity. Administration of carbachol to the latter animals produced myoclonic responses. These results are discussed in relation to the inhibitory effect of Ruthenium red on Ca2+ transport and binding to membranes, and to the role of this cation on neurotransmitter release.

Animals↗
PubMed1979

Physiological dissection of various effects of ruthenium red dye on Paramecium cells.

Polycationic dye ruthenium red, but not alcian blue, if externally applied to Paramecium cells quickly inhibits their phagocytosis. Ruthenium red combined with the cell surface diminishes frequency and duration of ciliary reversals and gradually inactivates the Ca++ gating mechanism. This effect persists for 1-3 after ruthenium red removal from the culture medium.

Animals↗
PubMed1979

Transmitter release: ruthenium red used to demonstrate a possible role of sialic acid containing substrates.

1. The possible function of sialic acid-containing substrates (SACS) in synaptic terminals of Aplysia was studied by intracellular injection of ruthenium red and of neuraminidase. 2. Ruthenium red, a dye known to have sialic acid as a molecular target, blocked transmission irreversibly in both cholinergic (buccal ganglion) and non-cholinergic (cerebral ganglion) synapses. 3. An intracellular site of action is likely because much less ruthenium red was necessary to block transmission when it was injected intracellularly than when it was presented by bath perfusion. 4. Ca2+ spikes recorded in the presence of tetrodotoxin or in Na+-free solution were not modified by ruthenium red or neuraminidase injections or perfusions. It is therefore improbable that these substances blocked transmission by blocking voltage-dependent Ca2+ influx. 5. Strong electrotonic depolarization of a pre-synaptic interneurone in the presence of 10(-4) M-tetrodotoxin caused a sustained post-synaptic response, which was abolished by ruthenium red. This result eliminates axonal conduction block as the principal mechanism of ruthenium red action. 6. Post-synaptic responses to ionophoretically applied acetylcholine (ACh) were not modified by bath perfusion of 2 x 10(-2) M-ruthenium red. 7. Biochemical analysis of pools of [3H]ACh was performed after injection of a precursor, [3H]acetate, into an identified interneurone. Ruthenium red appeared to increase significantly the 'free' (cytoplasmic) ACh pool without any change of 'bound' (vesicular) [3H]ACh-pool. 8. A model is proposed in which SACS act as intracellular Ca2+ receptors involved in transmitter release.

Action Potentials↗
PubMed1979

Ruthenium red as a stain for electron microscopy. Some new aspects of its application and mode of action.

Commercial ruthenium red has been tested for its purity by spectrophotometry. Impurities detected by this method could be abolished by nitric acid-precipitation of ruthenium brown. This substance has no effect on cell surface staining and converts almost completely to ruthenium red under the conditions used in electron microscopy. It was found, by photometric analysis, that in the ruthenium red-osmium tetroxide-cacodylate combination, generally used for cell surface staining, chemical reactions between ruthenium red and osmium tetroxide occur. As aerial oxidation of hexammineruthenium2+ leads to a product with some surface staining capability, it is suggested that an oxidized product of ruthenium red is responsible for binding to cellular components, and that a reduced product of osmium tetroxide gives an additional contrast enhancement. In ruthenium red-osmium dioxide combinations ruthenium red seems to bind to cell surfaces without any molecular alteration, and contrast is gained by the model proposed by Blanquet (1976b). The latter method could open a way for investigating the binding of ruthenium red to certain natural compounds involved in calcium transport, as postulated by a number of authors. Both ruthenium-osmium combinations differ in their cell surface staining ability. The ruthenium red-osmium dioxide combination tends to form distinct subunits, whereas the osmium tetroxide variety stains homogeneously. In combination with osmium dioxide, the surface staining is affected by EDTA, and, in contrast to osmium tetroxide, a successive application of ruthenium red and osmium dioxide as possible.

Blood Platelets↗
PubMed1977

The appearance of the outflow apparatus of the eye after staining with ruthenium red.

The outflow apparatus from adult baboon and rabbit eyes was stained with the inorganic dye ruthenium red. The ruthenium reaction product coated the surface of the trabecular meshwork cells and the canalicular endothelial cells. Deposits also impregnated the various connective tissue elements within the trabeculae and the extracellular spaces of the endothelial meshwork. A fine fibrillar network could also be identified with ruthenium red and this was present in the trabecular cores and the extracellular spaces of the endothelial meshwork. It was considered that the fibrillar network may represent a matrix of glycosaminoglycans and glycoproteins. The significance of these materials in relation to aqueous outflow was discussed.

Animals↗
PubMed1979

The interaction of ruthenium red with surface charges controlling excitation-contraction coupling in frog sartorius.

Frog sartorii were incubated in choline Ringer solution containing different amounts of the cationic dye ruthenium red, and were subsequently superfused with ruthenium red-free solution. The contraction threshold was measured during and after the incubation at different calcium and magnesium concentrations. During incubation in ruthenium red the threshold potential is slowly shifted to more positive values depending on time of incubation and the ruthenium red concentration (10--300 microns). After ca. 40 min of incubation a saturation potential is reached. The threshold shift is already maximal (-38mV threshold potential) at 30 microns of ruthenium ret regardless of the calcium concentration up to 5 mM. Omitting calcium from the incubation solution or adding 0.5 mM magnesium instead of calcium resulted in a more negative saturation potential (-48 mV). Washing the muscle in ruthenium red-free solution for 60 min after the incubation fails to reverse the threshold shift completely. The irreversible component of the threshold shift does not depend on the divalent cation concentration during incubation as long as the saturation value during incubation is more positive than -50 mV. The contraction threshold achieved after incubation with ruthenium red is dependent on the divalent cation concentration with calcium being twice as effective as magnesium. The effect of ruthenium red is greatest at small divalent cation concentrations and not significant at 50 mM. Incubating muscles with 5 units of neuraminidase shifted the concentration threshold to more positive potentials to the same extent as incubation with ruthenium red. Subsequent treatment of the neuraminidase-treated muscles with 30 microns of ruthenium red has no further effect on contraction threshold. The alternative experiment, first incubation with ruthenium red and then treatment with neuraminidase, gives the same results. The results are explained by the interaction of ruthenium red with membrane-bound sialic acid. This interaction is thought to result in a decrease in negative charges which results in a shift of the surface potential and hence of the contraction threshold to more positive potentials.

Animals↗
PubMed1976

Electron microscopy of surface structures of Rickettsia prowazeki stained with ruthenium red.

In studying surface structures of Rickettsia prowazeki (E and Breinl strains) by ruthenium red staining, a microcapsular layer 125-165 A thick, composed of subunits 85--100 A in diameter with a periodicity of 100--120 A as well as the inner layer of the cell wall 40--60 A thick were clearly revealed. In tangential sections of cells, subunits of the microcapsular layer were found in parallel striation arrays. These structures presumably contain acid mucopolysaccharides detectable by ruthenium red staining. Besides, hitherto unreported intracytoplasmic membrane structures were detected in ruthenium red-stained rickettsiae.

Cell Membrane↗
PubMed1977

Age-related change in the neuronal microenvironment: penetration of ruthenium red into extracellular space of brain in young adult and senescent rats.

The volume of the extracellular space, which contributes to the microenvironment of neurons, is diminished in the brains of senescent (as compared to adult) rats and an age-related change in its composition has been hypothesized. To test this hypothesis we have compared the penetration of ruthenium red, a polyanion selectively distributed in the extracellular space, into the dentate gyri of young adult and senescent Fischer 344 rats. Slices of hoppocampal formation were fixed by immersion, first in a glutaraldehyde solution containing ruthenium red, then in a solution of osmium tetroxide containind examined by electron microscopy. Dense particles of ruthenium red reaction product were readily localized in intercellular channels and synaptic clefts and the depth of penetration of ruthenium red in 25-month-old rats, as compared with 3-month-old animals, was found. These data indicate an age-related change in the charge density of the intercellular channels in the dentate gyrus of 25-month-old rats. They suggest a primary age-related change in the charg density of extracellular macromoledules, presumed to be primarily glycosaminoglycans, with a consequent change in water binding capacity and volume of the extracellular space.

Aging↗
PubMed1978

Electronmicroscopic histochemical investigation of the surface of the cell membrane of the renal tubules with the use of ruthenium red and lantanum nitrate.

The renal tubules were investigated with the use of ruthenium red and lanthanum nitrate to titrate the carbohydrate groups which are localized on the differentiated surface of the cell membranes and intercellular spaces. Ruthenium red visualized the surface coat which is tightly bound to the outer lamellae of the cell membrane. Lanthanum nitrate used in this investigations is a valuable marker of the intercellular spaces. The applied markers have visualized in the kidney the canals which are originated from the foldings of membranes of the tubular cells which adhere to the basal membrane. The markers used in combination with the fixative glutaraldehyde-paraformaldehyde give an electrondense envelope of the cell membrane which is more distinct as compared with specimen treated with glutaraldehyde and markers only.

Animals↗
PubMed1975

Distribution of ruthenium red staining material in the cerebral cortex of rat.

The ultrastructural distribution of ruthenium red staining material is studied in the cerebral cortex of rat. It is established that the material is located equally along the cell surface of the neurons, glial and endothelial cells, and within a number of cell organelles: mitochondria, smooth endoplasmic reticulum, synaptic vesicles and throughout both membranes of the nuclear envelope. Using digestion with hyaluronidase and trypsin, and lipid extraction an attempt is made to determine the participation of some chemical compounds such as acid mucopolysaccharides, glycoproteins, glycolipids and acid polypeptides in the composition of the ruthenium red-positive material. Moreover, the problem of the optimal conditions for the carrying out of ruthenium red staining in the CNS is discussed.

Animals↗
PubMed1975

The effect of ruthenium red and Concanavalin A on the vitelline surface of fertilized and unfertilized rabbit ova.

Fertilized and unfertilized rabbit ova exposed to ruthenium red with the zona intact or after mechanical removal, for the demonstration of mucopolysaccharides on the surface coat. Ova were also exposed to Concanavalin A. Ruthenium red bound strongly to elements of the zona and increased the opacity of the plasmalemma. There was no notable change in staining of the vitellus following fertilization. Although there were ruthenium red stained bodies resembling cortical granules in crypts in the plasmalemma, there was no evidence for cortical granule extrusion of mucopolysaccharides. The Concanavalin reaction was seen over both unfertilized and fertilized ova, demonstrating receptors for terminal alpha-D-mannopyranosyl, alpha-D-glycopyranosyl, beta-D-fructofuranyosyl residues. The deposit on the fertilized egg surface was enormously enhanced indicating a change in the distribution of receptor sites.

Animals↗
PubMed1975

Radioactive ruthenium red accumulation by tumors: a potential scanning agent.

The distribution of 103-Ru-Ruthenium red in tumour bearing animals indicates that Ruthenium red labelled with radioruthenium shows the characteristics of a tumour scanning agent. Its accumulation seems to be related to its specific binding to mucopolysaccharides. In addition, this radiocompound shows a potential value in the study of other pathological states involving the metabolism of mucopolysaccharides and glycoproteins.

Animals↗
PubMed1975

Mechanism of action of "ruthenium red" compounds on Ca2+ ionophore from sarcoplasmic reticulum (Ca2+ + Mg2+)- adenosine triphosphatase and lipid bilayer.

Sarcoplasmic reticulum (Ca2+ + Mg2+)-ATPase was previously shown to have Ca2+-dependent and -selective ionophoric activity when tested in oxidized cholesterol lipid bilayer membranes (Shamoo, A. E., and MacLennan, D. H. (1974) Proc. Natl. Acad. Sci. U. S. A. 71, 3522). ruthenium red, a known inhibitor of (Ca2+ + Mg2+)-ATPase, is found to inhibit the Ca2+-ionophoric activity associated with (Ca2+ + Mg2+)-ATPase. Furthermore, ruthenium red alone acts as an anion-selective ionophore in lipid bilayers with the the following selectivity sequence for anions: l- greater than Cl-, Br- greater than F- greater than NO3-. The PCl-/PNa+ ratio was approximately 4/l. The presence of ruthenium red in excess of Ca2+ ionophore in lipid bilayer experiments converts the cation selectivity of the bilayer due to Ca2+ ionophore into anion selectivity.

Adenosine Triphosphatases↗
PubMed1978

Effects of ruthenium red, A23187 and D-600 on steroidogenesis in Y-1 cells.

The effects of the calcium antagonists ruthenium red and D-600 and the cation ionophore A23187 on steroidogenesis were investigated. Steroidogenesis triggered by corticotrophin and cyclic AMP was inhibited by each of the agents. Incubation of Y-1 cells with an excess of ethyleneglycol-bis-(beta-amino-ethylether)-N,N'-tetraacetic acid (EGTA) abolished the steroidogenic response to corticotrophin while the response to cyclic AMP was unaffected. The ability of ruthenium red and D-600 (1 . 10(-5) M), and A23187 (6 . 10(-6 M) to inhibit a response which does not require the presence of extracellular calcium (cyclic AMP induced steroidogenesis) suggests that they are altering intracellular calcium. Neither of the calcium antagonists nor the cation ionophore inhibited the steroidogenic response to exogenous pregnenolone, thereby suggesting that the cells were still viable. Only when A23187 was used in the presence of a 15-fold increase in extracellular calcium (4.8 mM) was the response to pregnenolone diminished. The data are interpreted as a further indication that, in intact cells, intracellular calcium plays a role in the steroidogenic pathway.

Adrenocorticotropic Hormone↗