Single potassium channels in membranes of isolated mesophyll barley vacuoles

Summary Voltage-dependent K channels could be identified in on-cell and excised patch-clamp records on membranes of isolated plant cell vacuoles. The current through a membrane patch is dominated by a channel population with a conductance of about 121 pS in symmetrical 250mm KCl solution. The single channel adopts at least two conducting levels the 121-pS state being most frequently observed. The channel shows outward rectification, representing a cation flux into the vacuoles. The rectification appears to be caused by a vanishing open probability and a short channel lifetime at hyperpolarizing voltages. A selectivity ratio of potassium over sodium of about 6 was derived as an estimate. Occasionally, an additional population of K channels with a single-channel conductance of approximately 18 pS is observed. This channel type exhibits outward rectification as well.     

Conditional expression of RPA190, the gene encoding the largest subunit of yeast RNA polymerase I: effects of decreased rRNA synthesis on ribosomal protein synthesis.

The synthesis of ribosomal proteins (r proteins) under the conditions of greatly reduced RNA synthesis were studied by using a strain of the yeast Saccharomyces cerevisiae in which the production of the largest subunit (RPA190) of RNA polymerase I was controlled by the galactose promoter. Although growth on galactose medium was normal, the strain was unable to sustain growth when shifted to glucose medium. This growth defect was shown to be due to a preferential decrease in RNA synthesis caused by deprivation of RNA polymerase I. Under these conditions, the accumulation of r proteins decreased to match the rRNA synthesis rate. When proteins were pulse-labeled for short periods, no or only a weak decrease was observed in the differential synthesis rate of several r proteins (L5, L39, L29 and/or L28, L27 and/or S21) relative to those of control cells synthesizing RPA190 from the normal promoter. Degradation of these r proteins synthesized in excess was observed during subsequent chase periods. Analysis of the amounts of mRNAs for L3 and L29 and their locations in polysomes also suggested that the synthesis of these proteins relative to other cellular proteins were comparable to those observed in control cells. However, Northern analysis of several r-protein mRNAs revealed that the unspliced precursor mRNA for r-protein L32 accumulated when rRNA synthesis rates were decreased. This result supports the feedback regulation model in which excess L32 protein inhibits the splicing of its own precursor mRNA, as proposed by previous workers (M. D. Dabeva, M. A. Post-Beittenmiller, and J. R. Warner, Proc. Natl. Acad. Sci. USA 83:5854-5857, 1986).     

Ion channels activated by osmotic and mechanical stress in membranes of opossum kidney cells

Summary For patch-clamp measurements cultured kidney (OK) cells were exposed to osmotic and mechanical stress. Superfusion of a cell in whole cell configuration with hypotonic media (190 mOsm) evokes strong depolarization, which is reversible by returning to the isotonic bath medium. In the cell-attached configuration the exposure to hypotonic media evokes up to six ion channels of homogeneous single-channel properties in the membrane patch. Subsequently, the channels became activated after a time lag of a few seconds. At an applied membrane potential of 0 mV, the corresponding membrane current is directed inward and shows a transient behavior in the time range of minutes. In the same membrane patch these ion channels can be activated by application of negative hydrostatic pressure. The channel has a single-channel conductance of about 22 pS and is permeable to Na⁺ and K⁺ as well as to Cl^–. It is suggested that volume regulation involves mechanoreceptor-operated ion channels.     

Prediction of F3 row performance from F2 individual plant data in oats

Summary Parameters estimated from a Gardner-Eberhart analysis of the F₂ generation of a six-parent diallel in oats (Avena sativa L.) were used to compare methods for predicting the performance of F₃ row plots. The prediction methods were: (1) individual F₂ plant performance (F2I), (2) parent average plus F₂ plot deviations (PF2), (3) parent average plus weighted F₂ plot deviations (PF2P), (4) best linear unbiased prediction (BLUP) of parent average plus F₂ plot deviations (BPF2), and (5) BLUP plus weighted F₂ deviations (BF2). The F₂ single-plant traits used for prediction were biological yield to predict F₃ biological yield, whole plant and primary tiller grain yield for prediction of F₃ grain yield, and whole plant and primary tiller harvest index (HI) to predict F₃ HI. Prediction methods were evaluated by correlations between predicted and observed F₃ performance. Prediction methods and traits for which correlations were greater than for F2I included: BF2 for biological yield, PF2, PF2P and BF2 for whole plant grain yield, PF2, BPF2, and BF2 for primary tiller grain yield. None had a correlation significantly greater than F2I for either measure of HI, where heritability was large. PF2 is the recommended method for traits with low heritability because of its simplicity and because it had the largest or nearly the largest correlation for each of the yield traits. F2I is the recommended method for traits with larger heritability.Contribution No. 8821 of the U.S. Regional Pasture Research LaboratoryDeceased     

Suppressor factor secreted by T hybridoma established from peripheral blood lymphocytes of a bone marrow transplantation patient. I. Establishment of human T-cell hybridoma and partial characterization of suppressor factor

A stable human T-cell hybridoma was established by cell fusion between activated human peripheral blood lymphocytes from an allogeneic bone marrow transplantation patient and the JD1-17 cell line, a subclone of the human T leukemia Jurkat cell line. This hybrid clone 1-8, which bore the surface phenotype of suppressor cells (CD8+HNK1+), spontaneously secreted a factor which, at high dilutions, suppressed the responses of T and B cells induced by mitogens and alloantigens. This suppressor factor was found to be heat-resistant (56 degrees C, 30 min), stable at alkaline but not acid pH, unaffected by 2-mercaptoethanol, and sensitive to trypsin. Preparative isoelectric focusing revealed an isoelectric point of 5.35. The suppressor activity was selectively absorbed by blast T cells. By gel filtration on Sephacryl S-200 and HPLC, the suppressor activity was found in two peaks corresponding to 40-45 kDa (monomer) and 90-95 kDa (dimer).     

Characterization of a Ca-dependent maxi K channel in the apical membrane of a cultured renal epithelium (JTC-12.P3)

Summary A Ca and potential-dependent K channel of large unit conductance was detected in the apical membrane of JTC-12.P3 cells, a continuous epithelial cell line of renal origin. The open probability of the channel is dependent on membrane potential and cytoplasmic-free Ca concentration. At cell-free configuration of the membrane patch, the open probability shows a bell-shaped behavior as function of membrane potential, which decreases at larger depolarization. With increasing Ca concentration, the width of the bell-shaped curve increases and the maximum shifts into the hyperpolarizing direction. For the first time the kinetics of this channel was analyzed under cell-attached conditions. In this case the kinetics could sufficiently be described by a simple open-closed behavior. The channel has an extremely small open probability at resting potential, which increases exponentially with depolarization. The low probability induces an uncertainty about the actual number of channels in the membrane patch. The number of channels is estimated by kinetic analysis. It is discussed that this K channel is essential for the repolarization of the membrane potential during electrogenic sodium-solute cotransport across the apical membrane.     

Lack of evidence for liver lectins functioning as IgM or IgG-Fc-receptors

We have tested whether mannose- and galactose-specific lectins on liver cells are able to bind antibody-antigen complexes and thus function as Fc-receptors. Rat hepatocytes and liver sinusoidal cells were isolated by collagenase perfusion and differential centrifugation. Rat erythrocytes were coated with purified IgM or IgG from rabbits immunized with rat erythrocytes. Both IgM and IgG coated erythrocytes bound to liver macrophages but not to hepatocytes. The binding of IgM and IgG coated red blood cells to liver macrophages could not be blocked by potent inhibitors for mannose- and galactose-specific macrophage lectins such as mannan, D-mannose-bovine serum albumin, N-acetyl-D-galactosamine, D-galactose-bovine serum albumin, or asialofetuin. Although lectin activity is calcium dependent and trypsin sensitive neither condition blocked rosette formation between liver macrophages and opsonized erythrocytes. Thus mannose- and galactose-specific lectins are not involved in the sequestration of IgM- or IgG-antibody-erythrocyte complexes in the liver.     

Receptor-mediated particle uptake by liver macrophages. The galactose-particle receptor mediates uptake via coated and also non-coated structures

The endocytosis pathways of particles with terminal beta-D-galactosyl groups were studied in isolated rat Kupffer cells by electron microscopy. Colloidal gold particles of sizes 5, 17 and 50 nm were coated with asialofetuin (ASF) and isolated liver macrophages were allowed to bind (at 4 degrees C) or take up (at 37 degrees C) these ligands. Particles of all three sizes were bound via the galactose-particle receptor as shown by carbohydrate inhibition experiments and were ingested effectively. But, whereas ASF-gold particles of sizes 5 and 17 nm are taken up via the coated pit/coated vesicle pathway, the 50 nm particles are not. These enter the cell via non-coated endocytic vacuoles. All three particle sizes are transported to the same lysosomal compartment. These observations demonstrate that at least in macrophages one receptor is capable to mediate endocytosis via two different pathways depending on ligand size and/or valency.     

Die spektrale Transmission des dioptrischen Apparates von Aeschna cyanea Müll.

Zusammenfassung Die spektrale Transmission wurde mit dem UMSP (UniversalMicro-Spektralphotometer) im Bereich von 550–300 nm am fixierten (Carnoy) und frischen dioptrischen Apparat einzelner Ommatidien von Aeschna cyanea Müll. (Odonata) gemessen.Die Transmission der Kristallkegel und der frischen Cornea ist zwischen 550–370 nm nahezu konstant; bei kürzeren Wellenlängen des UV-Bereiches sinkt die Transmission etwas. Erst bei Wellenlängen, die kürzer als 330 nm sind, wird der Transmissionsabfall signifikant.

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The spectral transmission of the dioptric apparatus of aeschna

Summary Spectral transmission was measured by means of a Zeiss Universalmicro-spectralphotometer between 550 and 300 nm on fixed (Carnoy) and also on fresh cornea and crystalline cones of single ommatidia of Aeschna cyanea Müll. (Odonata).Both crystalline cones and fresh corneas (in contrast to fixed corneas) have nearly constant transmission from 550 to 370 nm; in uv-range with shorter wavelengths the transmission decreases slightly. Only at wavelengths shorter than 330 nm is the absorption significant.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.

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Mit Unterstützung der Alexander von Humboldt-Stiftung.