Search PubMed⌕ Search

Biomedical subjects

H Rahmann

Publications and source records attributed to H Rahmann.

At least 55 records · Page 3Linked to original sources

Histochemical investigations on the influence of long-term altered gravity on the CNS of developing cichlid fish: results from the 2nd German Spacelab Mission D-2.

The effect of long-term (10 days) altered gravitational conditions upon succinate dehydrogenase (SDH) reactivity in total brains as well as in individual brain nuclei of developing cichlid fish larvae had been investigated by means of semiquantitative histochemical methods (densitometric grey value analysis). Increasing accelerations from near weightlessness (spaceflight) via 1g controls to 3g hyper gravity (centrifuge) resulted in slightly increasing "all over the brain" (total brain) SDH reactivity. When focusing on distinct neuronal integration centers within the same brains in order to find the anatomical substratum of the gross histochemical data, significant effects of altered gravity only within vestibulum related brain parts were obtained.

Animals↗

Influence of altered gravity on the cytochemical localization of cytochrome oxidase activity in central and peripheral gravisensory systems in developing cichlid fish.

Cichlid fish larvae were reared from hatching to active free swimming under different gravity conditions: natural environment, increased acceleration in a centrifuge, simulated weightlessness in a clinostat and near weightlessness during space flight. Cytochrome oxidase activity was analyzed semiquantitatively on the ultrastructural level as a marker of regional neuronal activity in a primary, vestibular brainstem nucleus and in gravity receptive epithelia in the inner ear. Our results show, that gravity seems to be positively correlated with cytochrome oxidase activity in the magnocellular nucleus of developing fish brain. In the inner ear the energy metabolism is decreased under microgravity concerning utricle but not saccule. Hypergravity has no effect on cytochrome oxidase activity in sensory inner ear epithelia.

Animals↗

Imaging of thick sections of nervous tissue with energy-filtering transmission electron microscopy.

Electron microscopy of nervous tissue requires on the one hand nanometre resolution for the analysis of fine structures of nerve cell contacts, for instance synaptic vesicles, synaptic membranes and associated organelles. On the other hand, the visualization of the three-dimensional organization of nervous tissue on the level of dendrites and neurites is essential for the understanding of neuronal integration and also for a stereological evaluation of quantitative parameters such as size and shape of synaptic contact zones, number and distribution of synaptic vesicles, organization of cytoskeleton and distribution of organelles like mitochondria and endoplasmic reticulum. Therefore, it is necessary to have access to the fine structure and to the spatial organization within one sample. Energy-filtering transmission electron microscopy (EFTEM) enables the imaging of sections up to 1 micron thickness with a high resolution because the chromatic error due to inelastic scattering is avoided by selecting electrons within a restricted energy-loss range for imaging. The contrast depends strongly upon the section thickness, the selected energy range and the composition of the sample, so that optimum imaging conditions can be found for each object. Different staining protocols enable either a high global contrast or a selective staining of peculiar tissue properties. The three-dimensional organization can be visualized with stereopairs or with extended tilt series, which shows that characteristic structures as the synaptic junctions are detectable only within a narrow range of orientations to the electron beam. This is especially important for quantitative approaches with stereological tools which profit generally from the fact that a wide range of section thickness is available with EFTEM. EFTEM is therefore a powerful tool for the imaging of thick sections of biological materials with attractive possibilities of contrast tuning and advantages for stereological quantifications. The main benefit is the rapid and effective visualization of the three-dimensional organization of cells and tissues.

Animals↗

Comparative electronmicroscopical investigations on the influences of altered gravity on cytochrome oxidase in the inner ear of fish: a spaceflight study.

The regional metabolic activity in the otolithic sensory epithelia of the inner ear of a cichlid fish (Oreochromis mossambicus) was investigated on light- and electronmicroscopical level using the cytochemical method for detection of cytochrome oxidase activity. In adult animals a characteristic distribution of mitochondria with high enzyme activity was found in sensory and non-sensory cells of otolithic sensory epithelia, which was correlated with regions with a high energy demand. These findings were the basis for studies on the influence of long-term altered gravity conditions in developing larvae: hypogravity (10(-4) g in spaceflight), normal gravity (1 g in a centrifuge in space and 1 g on earth) and hypergravity (3 g in a laboratory centrifuge). Cytochrome oxidase activity was quantified in different parts of the sensory hair cell synapse in the vestibular sensory epithelia utricle and saccule: apical and basal cytoplasm, postsynaptic area of the afferent synapse and presynaptic region of the efferent synapse. Our results show that the energy metabolism of utricle, but not of saccule is decreased after microgravity exposure during the 2nd German Spacelab Mission D-2. However, a general effect of the spaceflight is detectable in both sensory epithelia. Long-term exposure to increased acceleration (3 g) had no effects on cytochrome oxidase activity in inner ear sensory epithelia.

Animals↗

An atlas of the distribution of NADPH-diaphorase in the brain of the highly derived swordtail fish Xiphophorus helleri (Atherinoformes:Teleostei).

The distribution of nicotinamide adenine dinucleotide phosphate-diaphorase (or -dehydrogenase, NADPHDH, a marker for nitric oxide synthase, NOS) positive neurons was demonstrated histochemically in the central nervous system of the swordtail fish Xiphophorus helleri, a highly derived telost of the atherinomorph outgroup. All nuclei of the swordtail fish, comprising almost all nuclei that have been described so far regarding the brains of telosts in general, were investigated. The results obtained clearly indicate, that NADPHDH positive neurons in the swordtail fish mostly are restricted to evolutionary primitive brain areas such as to most dorsomedial, dorsocentral and ventral portions of the telencephalon, to the preoptic area and to some thalamic, hypothalamic and pretectal nuclei of the diencephalon including the pituitary and the pineal organ, to the nuclei of the mesencephalic and myelencephalic cranial nerves, to the myelencephalic reticular formation and to the Mauthner cells. Some positive neurons were also observed within the mesencephalon, including the cerebellar body and its valve. Among non-neuronal cells, especially ependymocytes were strongly stained. It is the particular goal of the present study to provide a complete account on NADPHDH in the brain of a fish species, since, firstly, the NADPHDH/NOS system becomes increasingly important with regard to the understanding of the molecular basis of memory formation, and, secondly, fishes generally are more and more intensively studied concerning neurobiological approaches.

Animals↗

Correlation of altered gravity and cytochrome oxidase activity in the developing fish brain.

The mitochondrial enzyme, cytochrome oxidase, was localized cytochemically in the nucleus magnocellularis, a primary relay nucleus of vestibular information within the area octavolateralis in the fish brain. Cichlid fish larvae were analyzed after long-term exposure (9 days) to altered gravity situations: increased acceleration in a centrifuge (3 g) and near weightlessness during space flight. Controls (1 g) were reared under identical conditions in the centrifuge but without rotation on earth or with an acceleration resulting in gravity of 1 g in space shuttle. Quantification of highly reactive mitochondria reveals a correlation of gravity and cytochrome oxidase activity: low enzyme activity in respect to 1 g controls under near weightlessness conditions and an increased activity after hyper-g exposure in a centrifuge. This gravity effect on the energy metabolism of vestibular nuclei of developing cichild fish seems to reflect adaptational processes in response to gravity stimulation.

Animals↗

Unusual brain nucleus in the diencephalon of a perciform (nototheniid) fish from Antarctica.

This cytoarchitectonic analysis on the rostral diencephalon of antarctic perciform fishes confirms the previously established perciform diencephalic body plan. However, it resulted in the detection of an additional, large and conspicuous brain nucleus (unusual nucleus, UN) in the antarctic nototheniid Pleuragramma antarcticum. This nucleus has not been seen before in any other perciform. The UN is probably homologous to a visually related nucleus (named nucleus rostrolateralis and lateral entopeduncular nuclear area, respectively), that has so far only been found in two most primitive (osteoglosso-form) and one most highly derived (atherinoform) species of teleost. Our present results indicate that this brain region, which belongs to the ancient diencephalic organization of ray-finned fishes, is expressed in the brains of fishes due to their particular requirements regarding life styles rather than due to their phylogenetical relationships.

Animals↗

Comparative methodological investigations on the cytochemical localization of calcium in brain and inner ear of cichlid fish.

Four different methods for calcium precipitation are compared in the optic tectum and the inner ear of the cichid fish, Oreochromis mossambicus. Several parameters are investigated concerning their influences on the reaction product. Three procedures (bichromate, fluoride, and oxalate-pyroantimonate) produce fine-grained deposits, often flocculent in the latter method. The fourth method (potassium-pyroantimonate) generates predominantly coarse-grained reaction product. The calcium content of the deposits is always proven with energy-filtering transmission electron microscopy (EFTEM). In both tissues fine-grained reaction product is found in endoplasmic reticulum and synaptic vesicles, and in addition in some mitochondria and at the cytoskeleton. The coarse-grained deposits of the potassium-pyroantimonate method have a more unspecific distribution. This is the only method which produces extracellular deposits in the inner ear, whereas in the optic tectum extracellular precipitates are always present except with the oxalate-pyroantimonate procedure. Two factors have an influence on the reaction product: the duration of fixation and the type of resin. The prolongation of the fixation time up to 24 hours leads to an increase of the reaction product, which also becomes coarse-grained. These observations are corroborated by quantification with image analysis. Furthermore the use of an epoxy resin compared to acrylic resins decreases the amount of reaction product produced. We show that the application of several methods is meaningful in order to understand the calcium properties of the investigated tissue, but it is necessary to optimize a certain method for a given tissue.

Animals↗

Brain gangliosides and memory formation.

It is generally accepted that the process of molecular facilitation of neuronal circuits by means of stabilization of synaptic contacts represents the structural basis for memory formation. At the distinct zone of synaptic contact the following basic events occur: alterations of the electrical field strength at the outer surface of synaptic membranes, conformational changes of membrane-bound functional proteins (ion channels, ion pumps, receptors) and activation of second messenger cascade. Finally a trophic feed-back between synaptic terminals and their cell bodies through retro- and anterograde neuronal transport exists, which guarantees a stabilization of the newly formed synaptic connection. As a controlled Ca(2+)-exchange between the extracellular space (mM Ca(2+)-concentration) and the synaptoplasm (microM Ca2+) is essential for all these events, the present research concentrates on Ca(2+)-mediated primary messenger systems at the outer leaflet of synaptic membranes and on Ca(2+)-mediated modulatory mechanisms. These enable an always efficient electroresponsiveness although the environmental temperature might have changed. In this regard gangliosides being amphiphilic sialic acid containing glycosphingolipids, which are highly accumulated in complex composition in synaptic membranes play an important role. According to specific physico-chemical properties, they are assumed to fulfill the task of neuromodulators in connection with calcium, and thus contribute to the transmission and storage of information. The outcome of a series of experiments derived from neurology, ecophysiology, behavioral sciences, electron microscopy, biochemistry and physical chemistry give strong circumstantial evidence for this concept.

Animals↗

Development and altered gravity dependent changes in glucose-6-phosphate dehydrogenase activity in the brain of the cichlid fish Oreochromis mossambicus.

Glucose-6-phosphate dehydrogenase activity was studied in the brain of the cichlid fish Oreochromis mossambicus during early ontogenetic development. In general a slight but continuous decrease in enzyme activity was found (9.5 +/- 0.5 nmol substrate cleaved per mg protein and per min at developmental stage 13 [= 1 day post hatch at 28 degrees C] to a value of 7.9 +/- 0.6 in adult brain). In order to investigate the possible influence of altered gravity during early ontogenetic brain development, fish larvae were exposed to an increased acceleration of three times earth gravity (3 g) or to functional weightlessness in a fast-rotating clinostat for 7 days. A significant increase of brain G6PDH activity of approx. 15% was found after exposure to hyper gravity, whereas a significant decrease of the enzyme activity, approximately 10%, was detected following functional weightlessness in respect to the corresponding 1 g controls. Analyses concerning the regain of normal control enzyme activity of the larvae revealed dramatic fluctuations within the first 5 h after exposure to an increased acceleration of 3 g. Thereafter, between day 1 and day 3 after exposure, brain glucose-6-phosphate dehydrogenase decreased slowly. At day 3 after exposure no further differences of the hyper-g larvae compared to the controls were found. Only slight changes in total brain glucose-6-phosphate dehydrogenase activity occur during ontogenetic development of cichlid fish. This suggests that a more or less constant enzyme activity is important during brain development, but is reacting very sensitively to changes in the environmental factor gravity.

Acceleration↗

Influence of ambient temperature on content and composition of brain gangliosides in vertebrates.

The concentration and composition of brain gangliosides have been comparatively investigated by means of one- and two-dimensional high performance thin layer chromatography in several European fish species exhibiting a pronounced capacity for thermal adaptation. The values were compared with those of tropical and Antarctic fish as well as with homeo- and heterothermic mammals. The relative proportion of higher sialylated (more polar) ganglioside fractions was elevated in brains of fish (a) whose habitat was naturally cold or (b) that had been experimentally or seasonally adapted to a cold versus a warm habitat. Furthermore, in adaptation to the cold, more alkali-labile brain ganglioside fractions were found with the following sequence: Antarctic fish (53-67%), cold acclimated fish (45-51%), warm acclimated fish (38-44%), tropical species (< 35%), hamsters in torpor (9%), active hamsters (4%) and pig (3%). These results indicate a possible mechanism at a molecular level to keep the neuronal membrane functional, even under changing temperature conditions.

Adaptation, Physiological↗

Notes on the organization of the rostral diencephalon of the atherinomorph swordtail-fish Xiphophorus helleri.

By using conventional histological methods, the nuclear organization of the rostral diencephalon of the swordtail-fish, Xiphophorus helleri (Poecilidae, Atherinomorpha, Teleostei) was analyzed and compared with those from other teleost species. The subdivisions of the entopeduncular region, especially in their relative position and cytoarchitectonic structure, reveal clear differences between the various teleost orders. A new nucleus, the lateral entopeduncular nuclear area (leNA), found within this region, is described.

Animals↗

Characterization of sialyltransferase-IV activity and its involvement in the c-pathway of brain ganglioside metabolism.

To characterize the sialyltransferase-IV activity in brain tissues, the activities of GM1b-, GD1a-, GT1b-, and GQ1c-synthases in adult cichlid fish and rat brains were examined using GA1, GM1, GD1b, or a cod brain ganglioside mixture as the substrate. The GD1a-synthase activity in the total membrane fraction from cichlid fish brain required divalent cations such as Mg2+ or Mn2+ and Triton CF-54 for its full activity. The Vmax value was 1,340 pmol/mg of protein/h at an optimal pH of 6.5, whereas the apparent Km values for CMP-sialic acid and GM1 were 172 and 78 microM, respectively. Cichlid fish and rat brains also contained GM1b-, GT1b-, and GQ1c-synthase activities. The ratio of GM1b-, GD1a-, and GT1b-synthase activities in fish brain was 1.00:0.89:1.13, respectively, and in rat brain 1.00:0.60:0.63. Incubation of fish brain membranes with a cod brain ganglioside mixture, which contains GT1c, and [3H]CMP-sialic acid produced radiolabeled GQ1c. It is interesting that the adult rat brain also contains an appreciable level of GQ1c-synthase activity despite its very low concentrations of c-series gangliosides. The GD1a- or GQ1c-synthase activity in fish and rat brain was inhibited specifically by coincubation with the glycolipids that serve as the substrates for other sialyltransferase-IV reactions. Thus, the GD1a-synthase activity was inhibited by GA1 and GD1b, but not by LacCer, GM3, or GD3. In a similar manner, the synthesis of GQ1c was suppressed by GA1, GM1, and GD1b, but not by LacCer, GM3, or GD3.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effect of light stimulation on the cytochemical localization of cytochrome C oxidase in photoreceptor cells of cichlid fish.

Cytochrome oxidase activity was evaluated cytochemically in rod photoreceptor cells in the retina of the cichlid fish Oreochromis mossambicus after different stimulation protocols. The enzyme activity was assessed semiquantitatively by estimating the volume ratio of mitochondria classified according to the intensity of enzyme reactivity. Dark adaptation for 5 hr induced an increase of cytochrome oxidase activity both in vivo and in vitro, i.e., in isolated retinas. Short-term illumination (1 hr) of isolated retinas adapted previously in vivo to darkness caused a significant decrease of enzyme activity, whereas short-term darkening after in vivo light adaptation had no effect. Chemical stimulation for 15 min with increased K+ concentration (20 mM) reduced the enzyme activity, i.e., chemical depolarization did not have the same effect as depolarization induced by darkening. Significant changes in cytochrome oxidase activity were apparent within 1 hr of stimulation, so that this method for analysis of neuronal activity can be applied even in short-term experiments.

Animals↗

Altered gravity affects succinate dehydrogenase reactivity in specific nuclei of fish brain.

The effect of long-term (10 days) altered gravitational conditions upon succinate dehydrogenase (SDH) reactivity in total brain as well as in individual brain nuclei of developing cichlid fish larvae has been investigated by means of semiquantitative histochemical methods (densitometric grey value analysis). Increasing acceleration from near weightlessness (spaceflight) via 1 g controls to 3 g hypergravity (centrifuge) resulted in slightly increased total brain SDH reactivity. When focusing on distinct neuronal integration centres within the same brains in order to find the anatomical substratum of the gross histochemical data, significant effects of altered gravity on vestibulum-related brain parts were obtained. The total brain results may therefore represent the sum of such particular indirect effects but may also comprise in addition a non vestibular-related general and therefore direct influence of altered gravitational conditions, possibly on all cells.

Animals↗

Exogenously applied gangliosides (GM1, GD1a and Gmix) fail to facilitate the induction of long-term potentiation (LTP) in the slices of hippocampus and superior colliculus of the guinea pig.

To confirm the effect of gangliosides on the facilitation of the induction of long-term potentiation (LTP), slices of hippocampus and superior colliculus from guinea pig were prepared. One group of slices was incubated in the standard medium containing gangliosides (GM1, GD1a, or a mixture of gangliosides from bovine brain, each at a concentration of 70 microM) for 2 h. The other group of slices was incubated in the standard medium only. In both groups of slices, tetanic stimulation induced LTP in a similar manner. In low Ca2+ (1 mM) medium, LTP was not induced in either group of slices. Thus, the present experiments could not confirm previous reports indicating that exogenously applied gangliosides facilitate the induction of LTP.

Animals↗

[Brain gangliosides and memory formation].

It is generally accepted that the process of molecular facilitation (= Bahnung) of neuronal circuits by means of stabilization of synaptic contacts represents the structural basis for memory formation. On the basis of physiological, electron microscopic, biochemical, and physicochemical data, a concept is presented speaking in favor of the idea that interactions between brain gangliosides (= amphiphilic sialic acid-containing glycosphingolipids with peculiar physicochemical properties), calcium, and functional membraneous proteins (ion channels, ion pumps, receptors, kinases) play a crucial modulatory role in the transmission and storage of information.

Animals↗

Gravity effects on membrane processes.

Application of the Gouy-Chapman-Debye-Hückel (GCDH) theory to a model membrane in contact with electrolytes of various concentrations and composition predict density variations within an interfacial layer. Assuming that on cellular dimensions hydrodynamics can be applied (the objections are briefly discussed) two types of gravity effects can be defined, 1. convection along the surface of vertically oriented membranes and 2. surface potential variations by layer deformations at horizontally oriented membranes. Both effects should affect transport across the layer to the membrane surface and across the membrane. According to the theoretical predictions first experiments with gramicidin channels incorporated into artificial phosphatidylserine bilayer membranes show a significant difference in single channel currents in vertical and horizontal membranes. The complexity of biological membrane functions requires investigation of isolated membrane surface reactions and transport systems to study the gravisensitivity for each process separately.

Cell Membrane↗