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Biomedical subjects

H Rahmann

Publications and source records attributed to H Rahmann.

At least 109 records · Page 6Linked to original sources

Lead inhibits the voltage-activated calcium current of Aplysia neurons.

Lead is a potent and reversible inhibitor of the voltage-dependent calcium current of Aplysia neurons in a concentration range (1-1000 microM) similar to that which elicits toxic effects in man. The threshold for inhibition is 1 microM and the dissociation constant about 90 microM. The inhibition is due to reduction of the peak current amplitude, and not due to alteration of the voltage dependence of the activation or inactivation. The effect of lead is specific to the calcium current as the delayed rectifier potassium and the sodium currents are not affected by concentrations of lead (200 microM) which give near-maximal inhibition. The calcium current of Aplysia neurons appears similar to the current flowing through the mammalian L-type calcium channel. This suggests that inhibition of the investigated calcium channel may contribute to the toxic effects of lead in mammals.

Animals↗

Cytochemical localization of high-affinity Ca2(+)-ATPase activity in synaptic terminals.

High-affinity Ca2(+)-ATPase activity in the optic tectum of the brain of cichlid fish was cytochemically localized using cerium ions to precipitate phosphate. Activation of the enzyme with micromolar instead of millimolar calcium concentrations (i.e., physiological cytoplasmic instead of extracellular concentrations) resulted in intracellular localization of reaction product attached to the cytoplasmic side of plasma membranes and to synaptic vesicles. The plasmalemmal enzyme activity was concentrated in synaptic regions. Synaptic vesicles in some terminals exhibited high amounts of ATPase activity, whereas others were free of reaction product. By use of electron energy-loss spectroscopy (EELS) and electron spectroscopic imaging (ESI) techniques, even small amounts of cerium-containing precipitates could be analyzed and precisely localized. The cytochemical observations are in good agreement with biochemical findings and therefore indicate that the calcium pump of neuronal plasma membranes can be successfully localized.

Animals↗

Ultrastructural localization of gangliosides, calcium and a high-affinity Ca(2+)-ATPase in nerve terminals: a contribution to the possible functional role of gangliosides.

By means of newly developed electron microscopical techniques (electron spectroscopic imaging, ESI; electron energy loss spectroscopy, EELS; immunogold labelling) a specific accumulation of endogenous calcium within the synaptic cleft and a distinct localization of a high-affinity Ca(2+)-ATPase at the inner sides of the pre- and postsynaptic membrane of nerve cells from fish brain have been demonstrated. Additionally, a differentiation-dependent expression of polysialoganglioside epitopes on the outer surface of nerve terminals in clustered arrangements was demonstrated using their ultracytochemical detection by means of the monoclonal antibody Q211. These results which are in agreement with parallel biochemical investigations on modulatory effects of exogenous gangliosides on a high-affinity Ca(2+)-ATPase in the CNS of vertebrates support our hypothesis that Ca(2+)-ganglioside complexes act as modulators for the processes of synaptic transmission and long-term neuronal adaptations.

Animals↗

Comparative monolayer investigations of surface properties of negatively charged glycosphingolipids from vertebrates (gangliosides) and invertebrates (SGL-II, lipid IV).

The surface properties of four negatively charged glycosphingolipids from vertebrates, the sialo-glycosphingolipids (= gangliosides) GM1, GD1a, GT1b and a sulfo-glycosphingolipid (= sulfatide), and of the two negatively charged glycosphingolipids from lower invertebrates, the glucurono-glycosphingolipid Lipid IV and the aminophosphono-glycosphingolipid SGL-II were investigated in monolayers at the air/water interface. The molecular peculiarities under investigation were surface pressure (pi) and surface potential (delta V) which are described for Lipid IV and SGL-II for the first time. The surface pressure/area isotherms of all glycosphingolipids were typical of a liquid-expanded monolayer and, with the exception of SGL-II, exhibited a phase transition to a liquid-condensed state at surface pressures above 20 mN/m. The surface potential/molecular area data found for gangliosides in the closely packed state at pi = 30 mN/m (GM1: delta V = -17 mV; GD1a: delta V = -35 mV; GT1b: delta V = -39 mV) showed only a slight influence of the additional number of negatively charged residues. For Lipid IV, the surface behavior was very similar to GM1 both possessing one negative group per molecule, whereas in SGL-II also the surface potential data (delta V = +173 mV) were different compared with GD1a both possessing two negative groups per molecule. The addition of Ca2+ condensed the monolayers of all glycolipids and increased the potential in the direction to more positive values, but these findings were less effective in SGL-II films. On the basis of monolayer results presented here, in biological membranes of invertebrates especially Lipid IV might play a similar role as the ganglioside GM1 in vertebrate cells.

Acidic Glycosphingolipids↗

Possible involvement of polysialogangliosides in nerve sprouting and cell contact formation: an ultracytochemical in vitro study.

In Cichlid fish (Oreochromis mossambicus) primary cell cultures from whole brain and optic tectum, the differentiation-dependent distribution of polysialogangliosides on the outer cell surface has been followed on an ultrastructural level. For this, a two-step labeling technique with the monoclonal mouse antibody Q211, recognizing a polysialoganglioside-associated epitope, followed by a secondary IgM antibody, coupled to colloidal gold sols as an electron-dense marker, has been used. The gold grains are not uniformly distributed over the whole cell surface, but rather are clearly arranged clusters. In cells from freshly hatched larvae, both cell bodies and nerve fibers strongly exhibit the polysialoganglioside epitope on their surface. With progressing development, neuronal cell labeling is more and more restricted to nerve fibers and especially to cellular adhesion zones, including synaptic terminals, thus suggesting a functional involvement of polysialogangliosides in nerve sprouting and initiation of both cell-to-extracellular matrix and cell-to-cell contacts.

Animals↗

Effects of exogenous ganglioside and cholesterol application on excitability of Aplysia neurons.

The effects of pressure-ejected gangliosides GM1 and GMix ("Cronassial") and cholesterol dissolved in sea water on the electrophysiological characteristics of Aplysia neurons were studied using voltage-clamp recording techniques. Two types of electrophysiological effects were found. In about 5% of neurons brief pulses (0.1-0.2 sec) of GM1 or GMix elicited fast and large currents associated with an increase in membrane conductance and clear reversal potentials. These currents were similar to those elicited by common neuro-transmitters. Thus it appears that gangliosides may activate a membrane-bound receptor on at least some neurons. Most (about 85%) of the 121 neurons studied showed responses to longer pulses (1.0-2.5 sec) of gangliosides. These responses were much smaller, usually had a relatively slow component, and could be mimicked by application of cholesterol. The currents elicited were either inward or outward and were often biphasic, with an small initial outward component followed by a larger slow inward current. The responses often became larger upon repeated application at short intervals, and long periods of wash were required for recovery. This type of response appears to reflect changes in the electrical properties of the cell induced by incorporation of small amounts of gangliosides or cholesterol into the membrane.

Animals↗

Effect of calcium and temperature on mixed lipid-valinomycin monolayers. A comparison of glycosphingolipids (ganglioside GT1b, sulphatides) and phosphatidylcholine.

The influence of calcium and temperature on pure lipid (bovine brain PC, sulphatides, ganglioside GT1b), valinomycin and mixed lipid-valinomycin monolayers at the air/water interface was studied. In mixed films, evidence was found that the two components were miscible. On the other hand, at higher surface pressures, phase separation occurs in the cases of PC and sulphatides. Measuring the area requirement and the collapse pressure the stability of both lipid and the peptide was increased in particular due to ganglioside-valinomycin interaction. The addition of 10(-5) M calcium into the subphase at 20 and 37 degrees C and surface pressures of 10 and 20 mN/m led to a condensing effect in ganglioside mixtures, with formation of aggregates as indicated also by the nearly ideal behaviour of two component monolayers.

Calcium↗

Axonal transport of intraocularly injected [3H-Sph]-GD1a in the chicken optic system and the fate of the exogenous ganglioside distributed by blood.

Twelve-day-old chicks (White Leghorn) received an injection of 481 kBq (8.1 nmol) of [3H-Sph]-GD1a, which was labeled in its sphingoid, into the right eye. Structures of the injected and the non-injected (control) optic system (retinae, optic nerves, chiasm, optic lobes), the cerebrum, blood liver, kidney, and fly-muscle were analyzed 1, 4, 8 and 14 days later, with respect to total non-volatile radioactivity and to that bound to lower-phase lipids and gangliosides. It was demonstrated that exogenous [3H-Sph]-GD1a was taken up by the retina and mainly catabolized. 3H-label, reincorporated into the lower-phase lipids and gangliosides as well as authentic exogenous [3H-Sph]-GD1a were transported rapidly anterogradely in the entire optic system. [3H-Sph]-GD1a, distributed via the blood stream, was taken up by liver, kidney and muscle and was metabolized faster in these organs than in the retina. The cerebrum and the brain structures of the control optic system incorporated 3H-radioactivity to a much lower extent than the non-neural organs.

Animals↗

Seasonal variability of sialo-glycoconjugates in the brain of the Djungarian hamster (Phodopus sungorus).

The influence of season, photoperiod and ambient temperature on the content of proteins, sialo-glycoproteins and gangliosides and on the composition of gangliosides of three different brain regions (cortex, cerebellum and basalbrain) of the Djungarian dwarf hamster (Phodopus sungorus) had been investigated. Concomittantly changes in body wt and fur colouration were recorded. Dwarf hamsters living under natural photoperiod and ambient temperature conditions ("outside") showed a distinct annual cycle in body wt (summer: about 45 g; winter: about 25 g) and fur colouration (summer: dark grey; winter: whitish). Among the three brain regions the mean concentration of proteins ranged between 120 and 155 mg protein/g wet wt. The sialo-glycoprotein content varied between 260 and 410 micrograms NeuAc/g wet wt, and that of gangliosides between 800 and 1650 micrograms NeuAc/g wet wt. Seasonal fluctuations were not found. The composition of brain gangliosides remained uninfluenced throughout the year in the cerebellum, whereas seasonal variations were observed in cortex and basalbrain. Consequently the concentration ratio of the two major mammalian ganglioside fractions GD1a vs GT1b remained almost stable in cerebellum (0.3). In contrast to this the seasonal values of cortex and basalbrain changed from 0.6 and 0.8 in winter to 0.7 and 1.1 in summer. This indicated a higher polarity of the gangliosides in these brain regions during cold adaptation. The results are discussed with regard to modulatory functions of neuronal gangliosides for the process of synaptic transmission during seasonal adaptation.

Animals↗

Method for the OsO4-free preservation of brain tissue suitable for immunocytochemistry in TEM.

Following conventional OsO4-fixation and embedding of brain tissue in Epoxide resins for ultrastructural investigations, on the one hand, the wellknown good depiction of structures is obtained, on the other hand, however, according to these procedures a considerable loss up to 80% of glycolipid-bound and up to 50% of glycoprotein-bound neuraminic acid of important native compounds has to be put up with. The fixation of brain tissue by 1% glutaraldehyde and 1% paraformaldehyde in 0.1 mol/l Na-K-phosphate-buffer, followed by dehydration in acetone or ethanol at--25 degrees C and embedding in Lowicryl K4M preserves for instance amphiphilic glycolipids (gangliosides) to a high degree (95%) and in addition provides a good depiction of neuronal structures. The incubation of ultrathin sections of carp brain, treated as described above, with the specific antibody Q211 against polar gangliosides gave first evidence for a specific labelling of these amphiphilic glycosphingolipids and their localization within the brain tissue.

Animals↗

Influence of Ca2+ and temperature on the interaction of gangliosides with valinomycin in mixed monolayers at the air/water interface.

The effects of Ca2+ and temperature on mixed ganglioside-valinomycin-monolayers at the air/water interface were studied. Surface pressure-area isotherms of the pure gangliosides (GM1, GD1a) exhibited the typical monolayer characteristics. Pressure-area isotherms of the cyclodepsipeptide, valinomycin, were determined. In mixed monolayers, positive and negative deviation from the mean molecular area indicated the two components were miscible. Especially in GD1a mixtures, the addition of 0.01 mM calcium exhibited, with low molar fractions of valinomycin, a demixing effect in the direction of the phase separation of the components.

Calcium↗

Changes in brain gangliosides of the neotene and metamorphic (thyroxine-induced) newt axolotl (Ambystoma mexicanum).

Qualitative and quantitative changes in the concentration of proteins, sialoglycoproteins and gangliosides and in the composition of gangliosides in the brains of the neotene and the thyroxine-induced metamorphic newt axolotl (Ambystoma mexicanum) were investigated. During metamorphosis two polar gangliosides (GT1b and GQ1b) decreased by about 5% each. On the contrary GD1a increased to 10%. Another developmental trend was a slight increase of two other disialogangliosides (GD1b, GD2). Additionally, incorporation profiles (2-8 days) of 14C-N-Ac-mannosamine, the specific precursor for gangliosides, in the brain of neotene and metamorphic axolotls were followed giving evidence of significant changes in the sialoglycoconjugate metabolism of the central nervous system during metamorphosis of this newt.

Ambystoma↗

Brain gangliosides in monotremes, marsupials and placentals: phylogenetic and thermoregulatory aspects.

The concentration and composition of brain gangliosides of 17 mammalian species belonging to the subclasses of Prototheria (monotremes), Metatheria (marsupials), and Eutheria (placentals) were investigated. The mean concentration of brain gangliosides ranges from 525 to 610 micrograms NeuAc/g wet wt in monotremes, 445-900 micrograms in marsupials and from 630 to 1130 micrograms in the placentals. In the phylogenetic series of mammals, a decrease in the complexity of brain ganglioside composition becomes obvious: a drastic reduction in the number of individual ganglioside fractions particularly those of the c-pathway of biosynthesis, took place from the level of monotremes to that of the marsupials and placentals. In monotremes, marsupials and "lower" placentals (insectivores) the percentage of alkali-labile gangliosides is relatively low (between traces and 5%), whereas in the higher evolved mammals it amounts to about 20% of all gangliosides. The ratio of the contents of the two major mammalian ganglioside fractions GD1a and GT1b is generally in the range of 1.0 and even higher; in the heterothermic platypus from the monotremes and in hibernators among the placental mammals, however, it is much lower (about 0.8). These data support the hypothesis that the brain ganglioside composition not only depends on the phylogenetic level of nervous organization (cephalization) but is additionally correlated with the state of thermal adaptation.

Animals↗

Brain gangliosides in birds with different types of postnatal development (nidifugous and nidicolous type).

Developmental profiles of 14 different brain gangliosides were followed from the first day after hatching to the adult stage in two bird species representing different strategies of posthatch development: the nidifugous type (leaving the nest directly post-hatch, e.g. quail) and the nidicolous type (remaining for longer period in the nest, e.g. finch). In the zebra finch, parallel with a striking increase in ganglioside concentration, two main postnatal changes in the ganglioside composition occurred: after hatching, concomittantly to an increased outgrowth of nerve fibers and synaptogenesis, the polysialogangliosides GQ1b and GP decreased in favour of the less polar fractions GD1b, GD1a and GT1b. The second period of changes started with the onset of myelination and was characterized by an increase of GM1 and GM1'. The results obtained for quails were in close agreement with those of chicken, showing only slight postnatal changes due to the nearly completed morphological differentiation. These data show that gangliosides are useful biochemical markers for brain development, indicating successive periods of brain maturation by means of preferential biosynthesis of specific fractions regardless of the type of development.

Age Factors↗