Induction and ionic basis of slow wave potentials in seedlings of Pisum sativum L.

Slow wave potentials (SWPs) are transient depolarizations which propagate substantial distances from their point of origin. They were induced in the epidermal cells of pea epicotyls by injurious methods such as root excision and heat treatment, as well as by externally applied defined steps in xylem pressure (Px) in the absence of wounding. The common principle of induction was a rapid increase in Px. Such a stimulus appeared under natural conditions after (i) bending of the epicotyl, (ii) wounding of the epidermis, (iii) rewatering of dehydrated roots, and (iv) embolism. The induced depolarization was not associated with a change in cell input resistance. This result and the ineffectiveness of ion channel blockers point to H(+)-pumps rather than ion channels as the ionic basis of the SWP. Stimuli such as excision, heat treatment and pressure steps, which generate SWPs, caused a transient increase in the fluorescence intensity of epicotyls loaded with the pH-indicator DM-NERF, a 2',7'-dimethyl derivative of rhodol, but not of those loaded with the pH indicator 2',7'bis(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF). Matching kinetics of depolarization and pH response identify a transient inactivation of proton pumps in the plasma membrane as the causal mechanism of the SWP. Feeding pump inhibitors to the cut surface of excised epicotyls failed to chemically simulate a SWP; cyanide, azide and 2,4-dinitrophenol caused sustained, local depolarizations which did not propagate. Of all tested substances, only sodium cholate caused a transient and propagating depolarization whose arrival in the growing region of the epicotyl coincided with a transient growth rate reduction.     

Isolation and characterization of two cDNA clones encoding ATP-sulfurylases from potato by complementation of a yeast mutant

Sulfur plays an important role in plants, being used for the biosynthesis of amino acids, sulfolipids and secondary metabolites. After uptake sulfate is activated and subsequently reduced to sulfide or serves as donor for sulfurylation reactions. The first step in the activation of sulfate in all cases studied so far is catalyzed by the enzyme ATP-sulfurylase (E.C. 2.7.7.4.) which catalyzes the formation of adenosine-5′-phosphosulfate (APS). Two cDNA clones from potato encoding ATP-sulfurylases were identified following transformation of a Saccharomyces cerevisiae mutant deficient in ATP-sulfurylase activity with a cDNA library from potato source leaf poly(A)⁺ RNA cloned in a yeast expression vector. Several transformants were able to grow on a medium with sulfate as the only sulfur source, this ability being strictly linked to the presence of two classes of cDNAs. The clones StMet3-1 and StMet3-2 were further analyzed. DNA analysis revealed an open reading frame encoding a protein with a molecular mass of 48 kDa in the case of StMet3-1 and 52 kDa for StMet3-2. The deduced polypeptides are 88% identical at the amino acid level. The clone StMet3-2 has a 48 amino acid N-terminal extension which shows common features of a chloroplast transit peptide. Sequence comparison of the ATP-sulfurylase Met3 from Saccharomyces cerevisiae with the cDNA StMet3-1 (StMet3-2) reveals 31% (30%) identity at the amino acid level. Protein extracts from the yeast mutant transformed with the clone StMet3-1 displayed ATP-sulfurylase activity. RNA blot analysis demonstrated the expression of both genes in potato leaves, root and stem, but not in tubers. To the best of the authors' knowledge this is the first cloning and identification of genes involved in the reductive sulfate assimilation pathway from higher plants.     

Efficient generation of chimaeric mice using embryonic stem cells after long-term culture in the presence of ciliary neurotrophic factor

The aim of our study was to evaluate whether ciliary neurotrophic factor (CNTF) can substitute for leukaemia inhibitory factor (LIF) in maintaining pluripotential embryonic stem (ES) cells in culture. Two subclones of D3 ES cells were used to assess cell proliferation and differentiation in the presence of CNTF, LIF or Buffalo rat liver (BRL) cell-conditioned medium, or in the absence of exogenous differentiation inhibiting factors. ES cells maintained in medium supplemented with CNTF for up to four weeks were injected into blastocysts to investigate theirin vivo pluripotency in terms of chimaera formation. CNTF inhibited ES cell differentiation in a dose-dependent manner. The most effective concentration was 10 ng CNTF per ml of medium. The effects of CNTF on ES cell differentiation and proliferation were comparable to those of LIF at the same concentration. BRL cell-conditioned medium was less effective at preventing ES cell differentiation but induced their proliferation very markedly. Both ES cell clones efficiently formed chimaeras after long-term culture with CNTF as the only differentiation inhibiting agent. The ability of these ES cells to colonize the germ-line is the ultimate proof that CNTF can preserve the pluripotency of ES cells.     

The pericellular boundary layer modulates acetylcholine receptor stability in cultured myotubes

Degradation of acetylcholine receptors in cultured chicken myotubes was measured by release into the medium of radioactivity from 125I-labeled alpha-bungarotoxin. Disturbance of the pericellular boundary layer by stirring of the culture medium shortened the half-life of receptor in the membrane from 24 to 12 h. The effect could not be explained by dissociation of toxin-receptor complexes or by conditioning of the bulk phase of the medium. The rates of synthesis and degradation of total cell protein and the degradation of lactoperoxidase-iodinated surface protein were not affected by medium stirring. The loss of glucosamine-labeled material from the cells was enhanced by stirring, however, and this resulted entirely from the increased shedding of high molecular weight glycosubstances from the cells. Cells in stirred cultures contained lower levels of surface coat material stainable with colloidal thorium. These results indicate that glycosubstances of the pericellular matrix protect ACh receptors from degradation.     

Evolutionary changes in non-histone chromosomal proteins within the Drosophila melanogaster group revealed by monoclonal antibodies

Monoclonal antibodies directed against nonhistone chromosomal proteins of D. melanogaster were tested for crossreactivity with the homologous antigens of various Drosophila species. — By indirect immunofluorescence it could be shown that three antibodies react only with polytene chromosomes of species of the D. melanogaster subgroup, and only much less with chromosomes of other species of Drosophila. — With chromosomes of various other species of the Sophophora or Drosophila radiations only a reaction at background level could be observed. — The results suggest that the three antibodies react with different antigenic determinants of a single protein whose conformation changed rather fast during evolution of the Drosophilidae.     

Ein Modell zur örtlich-zeitlichen Signalübertragung im visuellen System des Menschen auf der Basis der linearen Systemtheorie kontinuierlicher Medien

Zusammenfassung Als Modell für die Übertragung örtlich-zeitlich variabler Helligkeitsmuster durch das visuelle System des Menschen wird ein dreidimensionales, homogenes neuronales Geflecht betrachtet. Die Ausbreitung der neuronalen Erregung erfolgt anisotrop in afferenter und lateraler Richtung und wird durch eine modifizierte Wärmeleitungsgleichung beschrieben, deren Hauptmerkmal ein regenrativer Signalanteil ist. Die zeitabhängige Lösung beschreibt u.a. die zeitliche Übertragungscharakteristik für flimmernde Gitter im gesamten Frequenzbereich. Die stationäre Lösung beschreibt die örtliche Übertragungscharakteristik für Gitter, wobei auch die Struktur rezeptiver Felder mit inhibitorischem Umfeld deutlich erkennbar ist. Die Struktur der Lösung läßt verstehen, wie sich mit zunehmender Tiefe im neuronalen Geflecht der Übergang von der Ortsselektivität zur Ortsfrequenzselektivität vollzieht.

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A model for the spatial-temporal signal transmission properties of the human visual system based on the linear system theory of continuous media

In this paper, a three dimensional homogeneous neural web serves as a model for the transmission of spatial-temporal luminous pattern by the human visual system. The propagation of neuronal excitation in afferent and lateral direction is assumed to be anisotropic. It is described by a modified heat conduction equation a main feature of which is a regenerative signal component. The time-dependent solution describes f.e. the temporal transmission characteristics for flickering grids in the whole frequency range. The stationary solution contains the spatial transmission characteristics for grids as well as the structure of receptive fields with inhibitory surrounds. The structure of the solution makes it under-standable how the transition from spatial selectivity to spatial-frequency-selectivity develops with growing depth of the neuronal web.

     

Derivative chromosomal structures from a ring chromosome 4

Summary This report describes the results from cultured lymphocytes studied at metaphase, anaphase, and interphase from an individual with a ring chromosome 4. A ring was present in 90.1% of metaphases. Special attention was directed towards the occurrence of derivative chromosomal structures, such as partially duplicated and triplicated rings, tricentric rings, chains of 3 interlocked rings, rod-shaped chromosomes, pulverized rings, and others.The clinical features of the individual (small stature and impaired mental development, hypoplastic thumbs, ptosis palpebrae hypoplastic external male genitalia, abnormal dermatoglyphic pattern) did not conform to a specific phenotype.

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Zusammenfassung Die Lymphocyten eines Patienten mit Ringchromosom 4 wurden in der Meta-, Ana-und Interphase beobachtet. 90,1% der Metaphasen wiesen einen Ring auf. Besondere Aufmerksamkeit wurde abnormen, aus dem Ring entstandenen Chromosomen zugewandt. Es fanden sich u. a.: partielle Doppel-und Dreifachringe, Ringe mit 3 Zentromeren, Ringe aus 3 miteinander verketteten Ringen, stabförmige Chromosomen und pulverisierte Ringe.Die Symptome des Patienten (Minderwuchs, Schwachsinn, Hypoplasie der Daumen, Ptosis palpebrae, Hypoplasie des äußeren genitale, abnorme Dermatoglyphen) waren zum größten Teil unspezifish.

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This work was supported in part by grants from the Deutsche Forschungsgemeinschaft and is part of a thesis by R.N. for a medical doctorate.     

Developmental changes of nerve growth factor and its mRNA in the rat hippocampus: Comparison with choline acetyltransferase

Previous experiments have demonstrated that in the septo-hippocampal system choline acetyltransferase (ChAT) is induced by nerve growth factor (NGF) (Gnahn et al. (1983) Dev. Brain Res. 9, 45-52) and that hippocampal NGF and mRNANGF levels are correlated with the density of cholinergic innervation (Korsching et al. (1985) EMBO J. 4, 1389-1393). In the present investigation we have compared the developmental changes of ChAT, NGF, and mRNANGF levels in this system. During the postnatal development of the hippocampus the time courses of NGF and ChAT were well correlated including the most rapid increase between P12 and P14. This increase in hippocampal NGF was preceded by a corresponding increase in mRNANGF. The developmental changes in hippocampal NGF levels were also closely reflected by corresponding changes in the septum. This, together with previous observations (Korsching et al., 1985) that the adult septum, in spite of relatively high NGF levels, does not contain measurable quantities of mRNANGF, suggests that the NGF levels in the septum are determined by the quantity of NGF transported retrogradely from the field of innervation rather than by local synthesis. During the prenatal period hippocampal NGF levels were relatively high, whereas the mRNANGF was below the level of detection. Since the ingrowth of septal fibers, and with that also the removal of NGF by retrograde transport, begins around birth, the relatively high prenatal NGF levels probably result from an accumulation produced by a small copy number of mRNANGF prior to the removal of NGF by retrograde axonal transport. It is concluded that the correlation of the developmental changes in NGF and mRNANGF with the ChAT activity in the hippocampus further supports the concept of a physiological role of NGF in the central nervous system.     

A satellite cell mitogen from crushed adult muscle

Single fiber-satellite cell units from skeletal muscle of adult rats were used to study the regulation of satellite cell proliferation. The satellite cells remained quiescent during culture in serum-containing medium but could be induced to enter the cell cycle by exposure to a saline extract of crushed adult muscle. The activity in the extract has a molecular weight greater than 30K and is heat and trypsin sensitive. The mitogenic activity does not result from transferrin. Little or no activity was obtained from crushed extracts of heterologous tissues. Proliferation of myogenic cells from rat embryos was also stimulated by the muscle mitogen but growth of muscle fibroblasts was not enhanced. The time response of satellite cell proliferation after exposure to the muscle mitogen showed that the cells enter DNA synthesis after a lag period of 18 hr and proliferate with a generation time of 12 hr. This confirms that satellite cells in adult muscle are in G0, or an extended G1. The mitogen is also effective in stimulating muscle growth and myoblast fusion in vivo when injected into 1-week-old rat pups. These experiments suggest that muscle regeneration is initiated by the release of an endogenous mitogen from traumatized muscle.