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  • 11
    ISSN: 1432-2048
    Keywords: Binding loci (elicitor) ; Elicitor ; Epipolarization microscopy ; Protoplast (parsley, soybean)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We describe a method which allows the visualization of elicitor-binding loci at the surface of plant protoplasts. Prerequisites for this method are the preparation of protoplasts under conditions of minimal proteolysis, and the availability of antibodies against either the elicitor itself or against the fluorescein portion of elicitor-fluorescein conjugates. Silver enhancement is used to amplify the visibility of 5-nm gold particles which are attached to an appropriate secondary antibody. Bound elicitor can then be visualized by epipolarization microscopy. This method, designated SEIG-EPOM (for silver enhanced immunogold as viewed by epipolarization microscopy), has been applied to protoplasts of parsley (Petroselineum cirspum L.), using the Phytophthora megasperma elicitor, and soybean (Glycine max Merr.) using polygalacturonic acid elicitor isolated from citrus pectin. We have been able to estimate the number of specific binding loci as being less than 100 per protoplast. Such loci possibly represent clusters of individual elicitor-receptor complexes. Structurally related elicitors have been shown to compete effectively for binding sites. The latter are sensitive to proteolysis, as is the elicitation response of protoplasts.
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  • 12
    ISSN: 1432-2048
    Keywords: Key words: Brassica (inflorescence) ; H+-pyrophospha-tase ; (immunogold detection) ; Plasma membrane ; Proton pumping ; Vacuolar H+-ATPase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Using a polyclonal antiserum specific for the tonoplastic H+-pyrophosphatase (tPPase), significant amounts of antigenic polypeptides of the correct molecular mass were detected in Western blots of plasma membrane isolated from cauliflower (Brassica oleracea L.) inflorescence by phase-partitioning and subsequent sucrose density centrifugation. Potassium iodide-stripped plasma membranes continued to give a strong positive signal, indicating that the PPase antigen detected was not a result of contamination through soluble PPase released during homogenisation. The same preparation contained negligible vacuolar (v)H+-ATPase activity and the A subunit of the vATPase could not be detected by immunoblotting. Plasma membrane fractions exhibited a proton-pumping activity with ATP as substrate, but such an activity was not measurable with pyrophosphate, although the hydrolysis of this substrate was recorded. By contrast, pyrophosphate supported proton pumping in tonoplast-containing fractions. Immunogold electron microscopy confirmed the presence of PPase at the plasma membrane as well as at the tonoplast, trans Golgi network, and multivesicular bodies. The density of immunogold label was higher at the plasma membrane than at the tonoplast, except for membrane fragments occurring in the lumen of the vacuoles which stained very conspicuously.
    Type of Medium: Electronic Resource
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Planta 141 (1978), S. 83-92 
    ISSN: 1432-2048
    Keywords: Cell wall regeneration ; Chlamydomonas ; Protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts from Chlamydomonas smithii prepared by the action of C. reinhardii gamete autolysine have been studied with respect to cell wall regeneration. “Natural” protoplasts within sporangia were also investigated for purposes of comparison. In both cases a new cell wall is completed within 2–3 h of the onset of regeneration. The first visible stages of wall regeneration are to be seen after 40–60 min as a fine fringe outside of the plasmalemma. The development of the typical “central triplet” follows within the next 1 h. Cell wall regeneration is reversibly inhibited by cycloheximide (10μg ml-1) and reversibly disturbed by concanavalin A (50 μg ml-1). Actinomycin D at concentration over 100μg ml-1 also inhibit but the inhibition is irreversible and peculiar membrane effects are observed. Chelators (ethylenediamine tetraacetic acid; ethyleneglycol-bis-aminoethyl ether) and 2-deoxyglucose slightly retard or have no effect on cell wall regeneration.
    Type of Medium: Electronic Resource
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  • 14
    ISSN: 1432-2048
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cytokinesis in the unicellular chlorococcalean alga Eremosphaera viridis de Bary has been investigated by electron microscopy of thin sections. The new plasmalemmata of the daughter cells in this organism form centrifugally within a phycoplast. Unlike other cell division systems each new plasmalemma is formed, not by the fusion of vesicles, but rather by the fusion of “open membranes” which are characteristically heavily stained. Measurements of these “open membranes” reveal that they are 11 nm thick with a central 4,5 nm unstained portion. The possible origin of these “open membranes” as burst-open vesicles has been suggested from the presence of intensely straining vesicles in the vicinity of the cell equator. Calculations of vesicle and “open membrane” surface areas support this contention.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Biological cybernetics 14 (1973), S. 71-83 
    ISSN: 1432-0770
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Computer Science , Physics
    Description / Table of Contents: Zusammenfassung 1. Es wird eine Folge von vier Modellen für das System der sakkadischen Augenbewegungen vorgeschlagen. Die Modelle werden von Stufe zu Stufe komplexer und bilden die experimentell gefundenen Antworten auf zunehmend kompliziertere Zielbewegungen nach. Bei der Konzeption der Modelle wird der Akzent auf Vereinbarkeit mit den strukturellen und funktioneilen Gegebenheiten der Neurologie gelegt. In jeder Stufe werden die Elemente dieser Modelle so gewählt, daß sie möglichst genau neuro-anatomischen Strukturen antsprechen und daß ihr Verhalten sich mit dem neurophysiologisch nachgewiesenen oder wahrscheinlich gemachten deckt. 2. Durch Ableitung von oculomotorischen Neuronen beim wachen Affen wurde die Dynamik des mechanischen Systems, bestehend aus den äußeren Augenmuskeln und dem Bindegewebe, in dem der Augapfel gelagert ist, erfaßt. Die Übergangsfunktion dieses Systems ist in die Modelle eingearbeitet. 3. Neuere Untersuchungen an den Strukturen des Hirnstamms, die für sakkadische Augenbewegungen verantwortlich sind, lassen im prämotorischen Apparat eine Anordnung vermuten, die im wesentlichen zwei neuronale Netzwerke enthält: einen Integrator und einen Pulsgenerator. Diese Schaltungen werden in den Modellen verwandt. 4. Nach Einarbeitung der obengenannten Änderungen in bestehende Modelle des sakkadischen Systems wird die Annahme, daß die visuelle Information durch das Nervensystem diskontinuierlich abgetastet wird, überflüssig. Die Modelle enthalten keine Abtastung, obwohl in Folge des sakkadischen Pulsgenerators ihr Verhalten als Ganzes noch immer das einer getasteten Regelung ist. 5. Das Grundmodell wird so modifiziert, daß sein Verhalten mit den Augenbewegungen übereinstimmt, die experimentell als Antwort auf kombinierte Sprung- und Rampenbewegungen des Ziels gefunden werden. Dies geschieht, indem der nach einer Reaktionszeit zu erwartende Fehler aufgrund des Momentanfehlers und seines Differentialquotienten geschätzt wird. 6. Parallele Datenverarbeitung ist eine allgemein bekannte Eigenschaft des Nervensystems. Durch Kombination dieser Eigenschaft mit einer zufälligen Entscheidungsschwelle wird das Modell so erweitert, daß es sich mit den experimentellen Befunden auch bei solchen Doppelsprüngen des Ziels deckt, bei denen der Sprungabstand kleiner als 0.2 sec ist. 7. Abschließend wird ein Modell vorgestellt, das ein Kontinuum von parallelen Datenverarbeitungskanälen einschließt und damit die retinotope räumliche Organisation des visuellen Systems sowie die tecto-bulbären motorischen Signale nachbildet. Es handelt sich hierbei um ein Denkmodell, das weder realisiert noch geprüft wurde. Es wird vielmehr dazu verwendet, komplexere Formen von Augenbewegungen zu diskutieren, wie sie z. B. aufzutreten scheinen, wenn der Entscheidungsprozeß zwischen den Hemisphären wechseln muß. Ebenso wird erörtert, wie das System schnelle Korrektursakkaden auslösen kann, deren Latenzen bis zu 85 msec kurz sind.
    Notes: Abstract 1. A sequence of four models is proposed for the saccadic eye movement control system. The models become increasingly complex as they are made to respond to increasingly more complicated target movements in accordance with experimental results. Compatibility with neurological structure and function is stressed in the formation of the models. In each case, the elements of the models are constructed to conform as closely as possible to neuroanatomical structures and behave in a way that has been established or suggested by neurophysiology. 2. The dynamic behavior of the mechanics of the extraocular muscles and eyeball suspensory tissues has been established by recording from oculomotoneurons in alert monkeys. The transfer function of this mechanical system is used in these models. 3. Recent experiments on the neural circuits in the brain stem that are responsible for saccadic eye movements suggest an arrangement of the premotor circuitry that contains two principal neural networks; an integrator and a pulse generator. This circuitry is used in the models. 4. When the above modifications are made to existing models of the saccadic system, they remove the necessity of supposing that the visual information is sampled by the nervous system. The models do not include a sampler although the saccadic pulse generator still makes the overall system behavior similar to that of a sampled-data system. 5. The basic model is modified to make its behavior agree with experimental eye movement responses to target ramps and step-ramps. This is done by using error and its rate of change to estimate the error that will exist one reaction time in the future. 6. Parallel processing of data is a well recognized property of the nervous system. By utilizing it in combination with a random decision threshold, the model is extended to produce results in agreement with experiments for double-step target movements in which the second step occurs less than 0.2 sec after the first. 7. Finally, a model is presented which incorporates a continuum of parallel processing to represent the retinotopic spatial organization of the visual system and the tecto-bulbar motor commands. The model is conceptual; it was not constructed or tested but is used to discuss more complex eye movement phenomena such as those that appear to occur when the decision process must shift between hemispheres and how the system might produce quick correcting saccades with latencies as short as 85 msec.
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  • 16
    Electronic Resource
    Electronic Resource
    Springer
    Biological cybernetics 49 (1983), S. 127-136 
    ISSN: 1432-0770
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Computer Science , Physics
    Notes: Abstract Single-unit recordings, stimulation studies, and eye movement measurements all indicate that the firing patterns of many oculomotor neurons in the brain stem encode eye-velocity commands in premotor circuits while the firing patterns of extraocular motoneurons contain both eye-velocity and eye-position components. It is necessary to propose that the eye-position component is generated from the eye-velocity signal by a leaky hold element or temporal integrator. Prior models of this integrator suffer from two important problems. Since cells appear to have a steady, background signal when eye position and velocity are zero, how does the integrator avoid integrating this background rate? Most models employ some form of lumped, oositive feedback the gain of which must be kept within totally unreasonable limits for proper operation. We propose a lateral inhibitory network of homogeneous neurons as a model for the neural integrator that solves both problems. Parameter sensitivity studies and lesion simulations are presented to demonstrate robustness of the model with respect to both the choice of parameter values and the consequences of pathological changes in a portion of the neural integrator pool.
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  • 17
    ISSN: 1432-0770
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Computer Science , Physics
    Notes: Abstract The discharge rates of premotor, brain-stem neurons that create eye movements modulate in relation to eye velocity yet firing rates of extraocular motoneurons contain both eye-position and eyevelocity signals. The eye-position signal is derived from the eye-velocity command by means of a neural network which functioins as a temporal integrator. We have previously proposed a network of lateral-inhibitory neurons that is capable of performing the required integration. That analysis centered on the temporal aspects of the signal processing for a limited class of idealized inputs. All of its cells were identical and carried only the integrated signal. Recordings in the brain stem, however, show that neurons in the region of the neural integrator have a variety of background firing rates, all carry some eye-velocity signal as well as the eye-position signal, and carry the former with different strengths depending on the type of eye movement being made. It was necessary to see if the proposed model could be modified to make its neurons more realistic. By modifying the spatial distribution of afferents to the network, we demonstrate that the same basic model functions properly in spite of afferents with nonuniform background firing rates. To introduce the eye-velocity signal a double-layer network, consisting of inhibitory and excitatory cells, was necessary. By presenting the velocity input to only local regions of this network it was shown that all cells in the network still carried the integrated signal and that its cells could carry different eye-velocity signals for different types of eye movements. Thus, this model stimulates quantitatively and qualitatively, the behavior of neurons seen in the region of the neural integrator.
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  • 18
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 164 (1989), S. 621-628 
    ISSN: 1432-1351
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A computer controlled setup is introduced which allows the song analysis of both male and femaleLeptophyes punctatissima during duetting in a laboratory situation. The essential acoustical parameters for the initiation of the male's phonotactic approach towards the stationary female are described. The female responds ‘reflex-like’ to the male song after a remarkably short delay time of about 28 ms. The male only performs phonotaxis if he perceives the female reply above an intensity value of about 50 dB SPL and if the female response falls within a critical ‘time window’ from 25 to a maximum of 55 ms after the onset of his song (Figs. 3 and 5). The sound intensity and overall time delay of the female response can be varied independently, so that the relationship between both parameters and their limitations for maximum phonotaxis distance can be described.
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  • 19
    ISSN: 1432-2048
    Keywords: Coated vesicle ; Cucurbita ; 1,3-β-Glucan synthase ; N-1-naphthylphthalamic acid binding ; Plasma membrane ; Vesicle, coated
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The two plasma-membrane (PM) markers: 1,3-β-glucan synthase and naphthylphthalamic-acid-binding capacity are localized in two peaks of activity on isopycnic Ficoll/D2O gradients prepared from a zucchini (Cucurbita pepo L.) hypocotyl post-microsomal fraction. The denser peak overlaps with the major distribution of clathrin and represents a region of the gradient enriched in coated vesicles (cv). Further purification of the pooled cv-fractions has shown that these PM marker activities are not borne by the cv, but are instead carried by smooth membrane fragments also present in these fractions. As judged from the results of Western blotting with polyclonal antibodies prepared against the 100-kilodalton (kDa) subunit of a PM H+-ATPase and the 70-kDa subunit of a tonoplastic H+-ATPase, these contaminants are of both PM and endomembrane origin. The PM contaminants however, differ from phase-partitioning- and free-flow-electrophoresis-purified PM prepared from microsomal fractions of zucchini hypocotyls in terms of their bouyant density in Ficoll/D2O gradients. Moreover, they do not appear to be present as sealed, outside-out, vesicles. Highly purified cv fractions from zucchini hypocotyls cross-react with subunit antibodies from both vacuolar and PM H +-ATPases. These results are discussed in terms of recent findings on cv ATPases from bovine brain.
    Type of Medium: Electronic Resource
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  • 20
    Electronic Resource
    Electronic Resource
    Springer
    Planta 193 (1994), S. 530-535 
    ISSN: 1432-2048
    Keywords: Amino acid ; Sucrose concentration ; Spinacia (leaf) ; Metabolite compartmentation ; Subcellular volumes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cellular and subcellular volumes in mature leaves of spinach (Spinacia oleracea L. US Hybrid 424) were determined stereologically from light and electron micrographs. Forty-nine-day-old leaves of spinach with a total leaf volume of 1177 μL per mg chlorophyll (Chl) were found to be composed of 3% epidermis, 58% mesophyll, 1% vascular tissue, 5% apoplasm and 32% gas space. In the epidermal cells 89% of the volume was occupied by the vacuole. The mesophyll cells consisted, expressed in mg·Chl−1, of 546 μL (79%) vacuole, 66 μL (9.5%) chloroplast stroma, 24 μL (34%) cytosol, 3.7 μL (0.5%) mitochondria and 2.1 μL (0.3%) nucleus. From previous measurements of the subcellular levels of sucrose, of phosphorylated intermediates of carbohydrate metabolism, of malate, oxoglutarate and various amino acids in illuminated leaves, and the above subcellular volumes, the corresponding subcellular metabolite concentrations have been determined. Of the substances measured, only with malate was the concentration higher in the vacuole than in the cytosol. The concentration of sucrose in the cytosol was 5 times, and that of amino acids even 30 times higher than in the vacuole.
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