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.     

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.     

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.     

Spontaneous cytotoxicity of macrophages against pancreatic islet cells

Activated peritoneal macrophages were found to lyse syngeneic [3H]leucine-labeled pancreatic islet cells or rat insulinoma cells after 15 h of coculture at 37 degrees C. Lysis was verified by electron microscopic analysis. Islet cell lysis was dependent on the T:E ratio and was comparable with P815 and L929 tumor cells used as targets. The cytotoxic activity was localized in the adherent fraction of Corynebacterium parvum activated peritoneal cells and was destroyed by incubation of cells with macrophage-toxic silica particles. Syngeneic thyrocytes and hepatocytes were found to be resistant to the cytolytic action of activated macrophages. It has been shown previously that macrophages contribute to pancreatic islet inflammation. The present in vitro analysis demonstrates that macrophages can function as effector cells in islet destruction.     

A method for measuring xylem hydraulic conductance and embolism in entire root and shoot systems

Current methods for determining the influence of xylem cavitationon hydraulic conductance are limited to unbranched stem or rootsegments with hydraulic conductances above c. 2 mmol s^–1MPa^–1. Lower conductances and/or highly branched systemsare encountered in seedlings, arid-land shrubs, herbs, and distalportions of shoot and root systems of trees. In order to quantifythe hydraulic impact of cavitation in such systems, existingtechniques have been modified. Branched shoot or root systemswere prepared for measurement by removal of leaves, or roottips, respectively. The shoot or root system was enclosed ina vacuum chamber with the proximal end protruding and suppliedwith perfusing solution. Flow through the xylem was inducedby chamber vacuum. Hydraulic conductance was determined fromthe slope of the flow rate versus pressure relationship. Xylemembolism was quantified from the increase in hydraulic conductancefollowing high pressure (100 kPa) perfusion of solution throughthe plant. Examples are provided of the application of the methodto cavitation studies in the cold desert shrub Artemisia tridentata. Key words: Hydraulic conductance, xylem cavitation, embolism, whole root/shoot system     

Molecular and genetic characterisation of German families with paramyotonia congenita and demonstration of founder effect in the Ravensberg families

Eighteen German families with a history of paramyotonia congenita (PC) were characterised by genetic und mutational analysis at the SCN4A locus, which encodes the -subunit of the adult skeletal muscle sodium channel. We concentrated our analysis primarily on these families to test the hypothesis that a predominance of one common mutation occurs in all German PC families and that this mutation arose in a common ancestor originating in the North-West of the country. The present eighteen PC families exhibit two different mutations (R1448C and R1448H) on various SCN4A dinucleotide repeat haplotypes and therefore the majority of the mutations probably occurred independently. However, the R1448H mutation is extremely frequent in the North-West of Germany (Ravensberger Land) on a specific SCN4A microsatellite haplotype, indicating a founder effect within this subpopulation. Our results suggest that the R1448C/R1448H mutations are by far the most common to be associated with the PC phenotype in the German population.This paper is dedicated to Professor Dr. Dr. Peter Emil Becker on the occasion of this 85th birthday     

A sensitive RNase protection assay using33P labeled antisense riboprobes

This article presents for the first time a modified protocol for RNase protection analysis that allows the substitution of³²P with³³P without loss of the high sensitivity of this method achieved with³²P. With this protocol, we were able to detect at least 1 pg of specific mRNA. In the RNase protection analysis³³P labeled riboprobes are more advantageous with regard to an easier handling and better resolution.