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M Lebert

Publications and source records attributed to M Lebert.

29 records · Page 2Linked to original sources

Behavioral mutants of Euglena gracilis: functional and spectroscopic characterization.

Three mutant strains of the phytoflagellate Euglena gracilis Z have been characterized in order to analyze the signal perception and signal transduction pathways involved in photo- and gravitaxis. Using the fluorescence of the chromophoric groups believed to be involved in photoperception (flavins and pterins) a method was developed for an in situ and in vivo detection of the paraxonemal body, the proposed location of the photoreceptor molecules. Two of the mutant strains, 1224-5/9f and 1224-5/1f, do not possess a stigma and also lack a paraxonemal body, as indicated by fluorescence measurements. The third strain, FB, has a small stigma, but only some cells contain a paraxonemal body. In contrast to the present hypothesis on photoorientation of Euglena, all strains were able to orient with respect to the light direction. However, the mutant strains did not show any orientation at low irradiances. At medium and high irradiances the strains 1224-5/9f and 1224-5/1f oriented perpendicular to the light direction (diaphototaxis) while cells of strains of FB showed partly negative phototaxis and partly diaphototaxis. Diaphototaxis was never observed in the wild type strain. Strains 1224-5/9f and 1224-5/1f showed normal graviresponses compared with the wild type. Astasia longa, a nonphtototactic relative of E. gracilis, as well as strain FB were both negative and positive gravitactic at all culture ages tested. This result confirmed the hypothesis that the paraxonemal body is not directly involved in graviperception.

Animals↗

Effects of hypergravity on the photosynthetic flagellate, Euglena gracilis.

Euglena gracilis, a unicellular, photosynthetic flagellate, orients itself by means of gravi- and phototaxis to reach and stay in regions optimal for survival and growth. An improved version of the slow rotating centrifuge microscope, NIZEMI, was used to test wild type and mutant strains for their responses to hypergravity. Wild type cells could actively move against the acceleration vector up to 8.5 gn and were centrifuged down at higher rates. Even at 10.5 gn, the highest value tested, cells were still negative gravitactically oriented as shown by video images. In contrast, all mutant strains as well as Astasia longa, a close relative of Euglena, could move against the acceleration vector under all conditions tested. With increasing accelerations the mean orientation of the populations shifted according to a vectorial addition of gravity and acceleration. The r-value, a statistical measure of the orientation of a population, increased with moderately increased acceleration rates and decreased at higher values. While wild type Euglena and two of the three mutant strains tested were exclusively negative gravitactically, in the third strain as well as in Astasia longa half of the population reacted negative gravitactically and the other half positive gravitactically. This variation of the wild type behavior was observed at moderate acceleration rates. At high accelerations the cells became exclusively positive gravitactic. The obtained results are discussed on the basis of the current model explaining gravitaxis.

Acceleration↗

Long-term cultivation of the flagellate Euglena gracilis.

Euglena gracilis, a unicellular photosynthetic flagellate, serves as a model system in signal transduction research. To further study its complex gravitaxis, experiments under microgravity are desirable. In preparation for long-term experiments on a space station, an autonomous cultivation unit has been developed and the culture conditions and surveillance methods have been established. The running time of more than 600 d under closed conditions with light as the only source of energy confirmed the stability of the Euglena population and gave new insights into its behavior. Physicochemical parameters such as oxygen concentration, temperature and pH as well as physiological parameters including cell density, motility, gravitactic orientation and pigmentation were recorded on a frequent basis. The suitability of the botanical bioreaction to serve as an oxygen supplier for animals in a closed system was demonstrated.

Animals↗

Disintegration of phycobilisomes in a rice field cyanobacterium Nostoc sp. following UV irradiation.

The effects of ultraviolet-B (UV-B, 280-315 nm) irradiation on survival, growth and phycobilisome assembly have been studied in the N2-fixing cyanobacterium Nostoc sp. Survival and growth were virtually arrested after 120 min of UV irradiation. The phycobiliprotein fractions showed a decrease in absorption and fluorescence as well as a shift towards shorter wavelengths indicating the impairment of energy transfer from phycobiliproteins to the photosynthetic reaction centers. This was further supported by SDS-PAGE analysis of the fractions which revealed a loss in high molecular mass rod-core and core-membrane linker polypeptides. Also the low molecular mass phycobilin (alpha beta) monomers decreased, showing that the supra-molecular organization of the phycobilisomes disintegrated during UV irradiation.

Bacterial Proteins↗

[Granulomatous endocarditis caused by streptococcus].

Two cases of granulomatous endocarditis are reported. The patients developed aortal endocarditis refractory to antibiotics. Therefore, aortal valve replacement was performed. In both cases, Streptococcus viridans was demonstrated in culture and bacterioscopically to be the cause of infection. Histological examination of the valves showed characteristic endocarditis with fibrinoid necrosis and histiocytic granulomas. Streptococci were found in the cytoplasm of macrophages. The possible causes of this special form of infectious endocarditis are discussed.

Adolescent↗

Characterization of Halobacterium halobium mutants defective in taxis.

Mutant derivatives of Halobacterium halobium previously isolated by using a procedure that selected for defective phototactic response to white light were examined for an array of phenotypic characteristics related to phototaxis and chemotaxis. The properties tested were unstimulated swimming behavior, behaviorial responses to temporal gradients of light and spatial gradients of chemoattractants, content of photoreceptor pigments, methylation of methyl-accepting taxis proteins, and transient increases in rate of release of volatile methyl groups induced by tactic stimulation. Several distinct phenotypes were identified, corresponding to a mutant missing photoreceptors, a mutant defective in the methyltransferase, a mutant altered in control of the methylesterase, and mutants apparently defective in intracellular signaling. All except the photoreceptor mutant were defective in both chemotaxis and phototaxis.

Bacterial Proteins↗

Methyl-accepting taxis proteins in Halobacterium halobium.

Methyl-accepting taxis proteins were identified and characterized in Halobacterium halobium, an archaebacterial species that is both chemotactic and phototactic. The data suggest direct involvement of methylation and demethylation in mechanisms of both chemotaxis and phototaxis and identify adaptation as the sensory process in which those reactions are likely to be involved. Analysis by electrophoresis and fluorography revealed methyl-accepting species, of apparent Mr between 90,000 and 135,000, that exhibited characteristics of sensory components. Those methyl-3H-labeled species were absent in a mutant blocked in taxis. Methylation of specific bands increased after positive chemostimuli and decreased after negative stimuli. Other methyl-3H-labeled bands, from 17 to 29 kd, exhibited features of biosynthetic intermediates, not of sensory components. Assay of rates of demethylation by measuring release of volatile forms of radiolabeled methyl groups revealed transient changes following chemo- or photostimuli that persisted for periods roughly equivalent to adaptation times. Negative chemostimuli induced increased rates of demethylation, as expected from fluorographic analysis, but positive chemostimuli also resulted in an increase. Photostimuli of either sign were followed by increases in rates of demethylation of shorter duration and lesser magnitude than chemostimuli-induced increases, a relationship that corresponded to differences in adaptation time.

Bacterial Proteins↗

Analysis of photoreceptor proteins of microorganisms by gradient gel electrophoresis and other biochemical separation methods.

Photoreceptor proteins for photoorientation in microorganisms are usually membrane bound and can be isolated by standard biochemical methods. Three examples are shown: the flagellates Euglena gracilis, Peridinum gatunense and the slime mold Dictyostelium discoideum. The photoreceptor of Euglena is attached to the basis of the flagellum and is composed of at least four chromoproteins which can be separated by gradient sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) fast protein liquid chromatography (FPLC) and isoelectric focusing (IEF); it contains pterins and a flavin as chromophoric groups. The photoreceptor of Peridinium absorbs in the red wavelength band. Though not yet identified in detail, multiple receptors are probably involved, as indicated by fluorescence spectroscopy. Dictyostelium shows positive and negative phototaxis in its amoebal form and exclusively positive phototaxis in its pseudoplasmodial form. It is still open to discussion whether the two stages use separate photoreceptors. From amoebae two photoreceptor pigments have been isolated, showing an absorption which resembles the action spectrum, one membrane bound with a molecular mass of 45 kDa and one cytoplasmic fraction with a molecular mass of 27 kDa.

Animals↗