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Biomedical subjects

F Wunderlich

Publications and source records attributed to F Wunderlich.

At least 73 records · Page 4Linked to original sources
PubMed1982

Intraerythrocytic development of Plasmodium knowlesi: structure, temperature- and Ca2+-response of the host and parasite membranes.

Thin-sectioning and freeze-etching electron microscopy were applied to explore the structure and the temperature- and Ca2+-response of the different host and parasite membranes during intraerythrocytic development of Plasmodium knowlesi in Macaca mulatta. The plasma membrane of uninfected erythrocytes is temperature- and Ca2+-responsive: chilling to 4 degrees C and exposure to 5 mM Ca2+ induces a slight decrease in IMP-frequency and the emergence of small IMP-devoid patches on P-faces. On parasite infection, the erythrocyte membrane becomes modified as indicated by an enhanced temperature-response and the appearance of caveolae, ca. 70-90 nm in diameter. The frequency of these caveolae is increased in schizont-infected erythrocytes. Moreover, electron dense plaques, ca. 40 nm in width, appear just beneath the erythrocyte membrane in late trophozoites and schizonts, thus indicating a further modification of the host cell membrane during parasite development. The membrane of the parasitophorous vacuole, derived from the host plasma membrane, dramatically reduces the IMP-frequency especially on the P-face upon parasite infection. This leads to an apparent reversal of the IMP-distribution persisting throughout the whole infection cycle. The parasite plasma membrane forms local compaction domains with the nuclear envelope in ca. 30% of the ring-stages and trophozoites, which disappear in late trophozoites and schizonts. Moreover, the IMP-frequency on plasma membrane fracture faces almost doubles during parasite development. Chilling induces a decrease in the IMP-frequency on P-faces of the plasma membrane. Surprisingly, however, the parasite plasma membrane and the vacuolar membrane respond to externally applied Ca2+ with almost a doubling of the IMP-frequency. The different parasite endomembranes also undergo characteristic changes during parasite development.

Animals↗
PubMed1980

In vitro ribosomal ribonucleoprotein transport upon nuclear expansion.

The interdependence of nuclear rRNA release and nuclear size is investigated in macronuclei isolated from Tetrahymena. Nuclei are induced to contract and to expand, without any structural disintegration of the nuclear envelope, by final Ca2+/Mg2+ (3:2) concentrations of 5 and 1.5 mM, respectively. Upon expansion, the average volume of nuclei increases from 600 +/- 42 to 811 +/- 76 micron3. Concomitantly, nuclei begin to release RNA following saturation kinetics. This RNA release stops immediately upon nuclear contraction. Similar to the in vivo situation, only advanced rRNA processing products are released in the form of ribosomal precursor particles, as identified in detail by polyacrylamide gel electrophoresis and rate zonal and isopycnic density gradient centrifugation. Three particle ty9es are released having average buoyant densities of 1.495, 1.470, and 1.532 g/cm3, exhibiting average sedimentation coefficients of 62, 62, and 35 S, and containing the immediate precursor to the 25S rRNA, 26S rRNA, and 17S rRNA, respectively. Tje rRNP release if ATP independent and noncoincident with the release of endogenous nuclear Pi, though it is Be2+ sensitive. Our data are compatible with the views that nuclear expansion is the prerequisite rather than the cause for the rRNP release and that nuclear pore complex associated ATPases play only, if at all, a minor role in nucleocytoplasmic exchange of rRNP.

Animals↗
PubMed1980

Nuclear envelope - plasma membrane - compaction domain in malaria parasites.

A novel compaction domain between the nuclear envelope and the plasma membrane is observed in intraerythrocytic Plasmodium knowlesi and Plasmodium chabaudi. The plasma membrane and the two nuclear membranes stick very closely together thus forming a heptalamina domain. This has an outer diameter of 14.3 +/- 1.1 nm and can be as long as 2/um. Fracture faces in this region appear to be devoid of the typical membrane-intercalating-particles. This compaction domain occurs only in ring-stages and early to mid trophozoites, but never in schizonts.

Animals↗
PubMed1978

Expansion and apparent fluidity decrease of nuclear membranes induced by low Ca/Mg. Modulation of nuclear membrane lipid fluidity by the membrane-associated nuclear matrix proteins?

Macronuclei isolated from Tetrahymena are contracted in form (average diameter: 10.2 micron) at a final Ca/Mg (3:2)concentration of 5 mM. Lowering the ion concentration to 1 mM induces an expansion of the average nuclear diameter to 12.2 micron. Both contracted and expanded nuclei are surrounded by a largely intact nuclear envelope as revealed by thin-sectioning electron microscopy. Nuclear swelling is accompanied by an expansion of the nuclear envelope as indicated by the decrease in the frequency of nuclear pore complexes from 52.6 to 42.1 pores/micron2 determined by freeze-etch electron microscopy. Contracted nuclear membranes reveal particle-devoid areas (average size: 0.21 micron2) on 59% of their fracture faces at the optimal growth temperature of 28 degrees C. About three-fifths of the number of these smooth areas disappear upon nuclear membrane expansion. Electron spin resonance using 5-doxylstearic acid as a spin label indicates a higher lipid fluidity in contracted than in expa,ded nuclear membranes. Moreover, a thermotropic lipid clustering occurs at approximately 17 degrees C only in expanded nuclear membranes. In contrast to the nuclear membrane-bound lipids, free lipids extracted from the nuclei rigidify with increasing Ca/Mg concentrations. Our findings are compatible with the view that the peripheral layer of the fundamental nuclear protein-framework, the so-called nuclear matrix, can modulate, inter alia, the lipid distribution and fluidity, respectively, in nuclear membranes. We suggest that a contraction of the nuclear matrix's peripheral layer induces a contraction of the nuclear membranes which, in turn, leads to an isothermic lateral lipid segregation within nuclear membranes.

Animals↗
PubMed1977

Effect of temperature on nuclear membranes and nucleo-cytoplasmic RNA-transport in Tetrahymena grown at different temperatures.

The effect of temperature on the nuclear envelope structure and the transport of total RNA and ribosomal subunits from nucleus to cytoplasm was examined in Tetrahymena cells propagated at two different temperatures. Freeze-etch electron microscopy of cells grown at 23 and 18 degrees C detects the emergence of smooth areas on the fracture faces of the nuclear membranes upon lowering the temperature below approximately 15 and approximately 12 degrees C, respectively. Coincident with these freeze-etch changes, a discontinuous decrease is observed in the nucleocytoplasmic RNA-transport; this is probably not due to a cease in RNA-synthesis. Below the thermotropic discontinuity observed in the transport of total RNA in 18 degrees-cells the nucleocytoplasmic transport of the small and large ribosomal subunits is equally retarded. Recent temperature studies on the endoplasmic reticulum membranes of Tetrahymena suggest that the freeze-etch changes in the nuclear membranes are induced by a thermotropic clustering of the membrane lipids. We conclude that this lipid clustering induces the permanent protein constituents in the nuclear envelope pore complexes to change from a relatively "open" into a relatively "closed" state thus causing the observed decrease in RNA-transport.

Biological Transport↗
PubMed1977

Reversibly contractile nuclear matrix. Its isolation, structure, and composition.

From Tetrahymena macronuclei we have isolated a reversibly contractile nucleo-skeleton, i.e., an "expanded" nuclear matrix which reversibly contracts when the total concentration of the bivalent cations, Ca and Mg (3:2), is decreased to 5 mM or increased to 125 mM. During contraction the average diameter of the expanded matrix becomes reduced by about 24%; this corresponds to a volume contraction of about 55%. The reversible contraction of the nuclear matrix does not depend on ATP and cannot be inhibited by salygran. The expanded matrix is obtained by removing carefully from the macronuclei 89.7% of the phospholipid, 99.6% of the DNA, 98.5% of the RNA, and 74.8% of the protein by treatment with Triton X-100 and digestion with DNase and RNase followed by an extraction with 2 M NaCl. Electron microscopy reveals, within the expanded matrix, residual equivalents to the structures characteristic for macronuclei: (a) a residual nuclear envelope with nuclear pore complexes; (b) residual nucleoli at the periphery; (c) a fibrillar internal network. The expanded matrix is essentially composed of proteins (96.2%) and traces of DNA (0.8%), RNA (0.5%), phospholipid (1.6%), and carbohydrates (0.9%). The last, which have been determined by gas chromatography, contain glucose, mannose, and an unidentified sugar in the ratio 1:5.4:5.7. The ratio of acidic to basic amino acids of the expanded matrix is 1.55. Sodium dodecyl sulfate (SDS) gel electrophoresis reveals a predominant protein with a mol wt of 18,000 which is apparently involved in the reversible contractile process. The mechanism of this reversible contraction of the expanded matrix remains to be elucidated, but it differs both from actin-myosin contraction systems and from the contractile spasmoneme system in vorticellids.

Calcium↗