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PubMed1997

[Analysis of the swimming pattern and the velocity of bacteria using video tracking method].

The swimming patterns and the velocities of several flagellated bacteria were measured by a computer assisted video tracking method. The moving path of the individual bacterium revealed that the bacterium frequently changed its swimming direction and velocity. The velocity among bacterial strains varies widely. In low viscous environment. Campylobacter jejuni has characteristic swimming pattern with frequent changes in their swimming direction. As the viscosity increase, C. jejuni increases its velocity at a little higher viscosity of 3 centipoise (cP) and secondly increases at about 40 cP. Different from other flagellated bacteria, the swimming pattern of C. jejuni in these two velocity peaks were changed. C. jejuni exhibited continuously forward moving path in the first peak, but in the second it repeated back and forth swimming pattern. We thus assumed that C. jejuni may use a different swimming mode in high viscous media from the original mode mediated by the propelling force of the flagella. This method is useful for a detail analysis of bacterial movement and moving patterns in different environmental conditions.

Bacterial Physiological Phenomena↗
PubMed2026

The genome sequence of Hyocomium armoricum (Brid.) Wijk & Margad. (Hypnales: Hypnaceae).

We present a genome assembly of Hyocomium armoricum (Flagellate Feather-moss; Streptophyta; Bryopsida; Hypnales; Hypnaceae). The genome sequence has a total length of 333.60 megabases. Most of the assembly (99.59%) is scaffolded into 11 chromosomal pseudomolecules. The mitochondrial sequence has a length of 104.43 kilobases and the plastid genome assembly has a length of 123.84 kilobases. This assembly was generated as part of the Darwin Tree of Life project, which produces reference genomes for eukaryotic species found in Britain and Ireland.

Flagellate Feather-moss↗
PubMed1996

Effect of microcystin on growth of single species and on mixed natural populations of heterotrophic nanoflagellates.

Natural populations of heterotrophic nanoflagellates were reduced in numbers and in situ growth rates during a toxic Microcystis bloom in a eutrophic lake. Microcystin was found in the particulate phase (algae) and in the dissolved phase (water) with maximum concentrations of 414 micrograms g freeze-dried algal material-1 and 141 micrograms l-1 lake water, respectively. An average reduction in growth rates of 49% was found when comparing the growth estimates before and after the toxic bloom had peaked. Similar reductions were found in laboratory experiments when growing mixed flagellate populations from two different systems with microcystin, but without predators. Concentrations of 10 micrograms toxin 1-1 reduced the growth rates by 36-41% (significantly different from the controls) and 1 microgram toxin 1-1 reduced the growth rates by 24-28% (not significant). Thus, natural populations of heterotrophic nanoflagellates seem very sensitive to microcystin. The growth characteristic of two cultured species was also tested in the presence of microcystin. Both the cultured species, Heteromita globosa and Spumella sp., grew well at 1-10 micrograms microcystin 1-1 and Spumella sp. was able to grow at 50 micrograms microcystin 1-1. However, the growth curves indicate that the decline in numbers during the stationary phase occurred faster in the presence of microcystin. The ecological consequences of a highly sensitive protozoan community may be that larger zooplankton species (copepods and cladocerans) are affected by reduced availability of food.

Animals↗
PubMed1996

Tubulin post-translational modifications in the primitive protist Trichomonas vaginalis.

Using several specific monoclonal antibodies, we investigated the occurrence and distribution of different post-translationally modified tubulin during interphase and division of the primitive flagellated protist Trichomonas vaginalis. Immunoblotting and immunofluorescence experiments revealed that interphasic microtubular structures of T. vaginalis contained acetylated and glutamylated but non-tyrosinated and non-glycylated [Brugerolle and Adoutte, 1988: Bio Systems 21: 255-268] tubulin. Immunofluorescence studies performed on dividing cells showed that the extranuclear mitotic spindle (or paradesmosis) was acetylated and glutamylated, which contrast with the ephemeral nature of this structure. Newly formed short axostyles also contained acetylated and glutamylated tubulin suggesting that both post-translational modifications might take place very early after assembly of microtubular structures. Our results indicate that acetylation and glutamylation of tubulin appeared early in the history of eukaryotes and could reflect the occurrence of post-translational modifications of tubulin in the primitive eukaryotic cells. These cells probably had a highly ordered cross-linked microtubular cytoskeleton in which microtubules showed a low level of subunit exchange dynamics.

Acetylation↗
PubMed1996

Centrin is a conserved protein that forms diverse associations with centrioles and MTOCs in Naegleria and other organisms.

Centrin, a approximately or equal to 20 kDa calcium-binding protein also known as caltractin, is a component of centrosome-associated algal flagellar roots capable of calcium-mediated contraction, and is also found in the centrosomes of vertebrate cells. Our analysis of a centrin gene from a protist, the amoeboflagellate Naegleria gruberi, reveals conserved features that distinguish centrins from calmodulin. Antibodies to bacterially expressed Naegleria centrin, which also recognize yeast Cdc31p, were employed to localize centrin immunoreactivity in selected organisms possessing specialized microtubule-organizing centers (MTOCs) or accessory structures. There is a striking morphological diversity of such structures. In the simplest associations, as found in Naegleria flagellates and vertebrates tracheal epithelium, centrin is intimately associated with the cylinder of the basal bodies. In cells with unfocused mitotic spindles, Naegleria amoebae and onion root tips, no localization of centrin was detected. In Dictyostelium discoideum and Saccharomyces cerevisiae, which lack centrioles, centrin immunoreactivity was observed as punctate cytoplasmic bodies but not associated with spindle pole MTOCs. In Paramecium multimicronucleatum, centrin immunoreactivity is localized to the infraciliary lattice, previously shown to exhibit calcium-mediated contraction. In Vorticella microstoma, known for the calcium-induced rapid contraction of its stalk, centrin immunoreactivity is localized to the contractile spasmoneme and myonemes. Similar antigens from Paramecium and Vorticella are detected by anti-centrin and anti-spasmin. The pattern of localization of centrin immunoreactivity supports the conjecture that a contractile system involving centrin, initially associated with centriolar structures, was recruited during evolution to build specialized organelles in different organisms and cell types.

Amino Acid Sequence↗
PubMed1997

SF-assemblin in Chlamydomonas: sequence conservation and localization during the cell cycle.

Previously, SF-assemblin has been identified as the filament-forming component of the striated microtubule-associated fibers (SMAFs), which emerge from the basal bodies in several green flagellates. We have sequenced cDNAs coding for SF-assemblin from Chlalmydomonas reinhardtii and C. eugametos. Comparison of the deduced amino acid sequences with the previously described green algal SF-assemblins shows identities between 54 and 71%, indicating a strong drift in sequence. Cells of C. reinhardtii were analyzed by double immunofluorescence using polyclonal anti-SF-assemblin and anti-alpha-tubulin. In interphase cells, SF-assemblin is associated with all four microtubular flagellar roots. During mitosis the SF-assemblin-based cytoskeleton is reorganized; it divides in prophase and is reduced to two dot-like structures at each spindle pole in metaphase. During anaphase, the two dots present at each pole are connected again. In telophase we observed an asymmetrical outgrowth of new fibers. These observations suggest a role for SF-assemblin in reestablishing the microtubular root system characteristic of interphase cells after mitosis.

Amino Acid Sequence↗
PubMed1998

SF-assemblin, striated fibers, and segmented coiled coil proteins.

The flagellar basal apparatus of many flagellate green algae contains noncontractile striated microtubule-associated fibers (SMAFs). The SMAFs consist of 2-nm protofilaments and are predominantly built from striated fiber (SF)-assemblin, an acidic 33-kDa protein. In this review we summarize the present knowledge concerning the biochemical properties of SF-assemblin and the molecular architecture of the SMAFs, provide evidence for homologous proteins and similar filament systems in other eukaryotes, and, finally, discuss possible biological functions of SF-assemblin.

Algal Proteins↗
PubMed1999

Locomotory waves of Koruga and Deltotrichonympha: flagella wag the cell.

We investigated the nature of the locomotory waves of Koruga and Deltotrichonympha, flagellates living symbiotically in the hindgut of the Australian termite Mastotermes darwiniensis. The locomotory waves consist of two components: metachronal waves of flagellar beating and undulations of the cell surface, which propagate synchronously with the same wavelength, frequency, and velocity. We asked, do body waves cause flagellar waves, or vice versa? Using video microscopy and selective inhibitors and drugs, we found that (1) the amplitude of flagellar waves remains constant independent of variations in the amplitude of body waves, (2) flagellar waves can occur in the complete absence of body waves, (3) flagellar waves can induce body waves on swollen regions, (4) inhibition of flagellar beating by dynein inhibitors causes disappearance of body waves, and (5) cytochalasin D induces changes in cell shape but does not inhibit locomotory waves. Therefore, flagellar waves are not produced passively by an active contractile system in the cell cortex; instead, metachronally beating flagella exert waves of pressure that induce passive undulations of a pliant cell surface. These results support Machemer's [1974] theoretical analysis of the data of Cleveland and Cleveland [1966: Arch. Protistenk. 109:39-63], who believed the opposite.

Animals↗
PubMed1979

Systemic leishmaniasis mimicking malignant histiocytosis.

A 22-year-old man with fever, hepato-splenomegaly and severe pancytopenia is described in whom the histologic features of the spleen, liver and lymph nodes were consistent with malignant histiocytosis. Characteristic Leishman-Donovan bodies were demonstrated on a bone marrow aspirate, and the diagnosis of visceral leishmaniasis was confirmed by culturing the flagellated forms of the pathogen, and by an excellent response to sodium stibogluconate. In view of the similarity in clinical and histologic appearance, visceral leishmaniasis must be considered in the differential diagnosis of malignant histiocytosis in geographic areas where leishmaniasis is still endemic.

Adult↗
PubMed2001

Applicability of growth rate, cell shape, and motility of Euglena gracilis as physiological parameters for bioassessment at lower concentrations of toxic substances: an experimental approach.

The responses of the growth rate, motility, and cell shape in the green flagellate Euglena gracilis to different concentrations of waste water substances from the pulp and paper industry were tested in a long-term study (7 days). Samples before (uncleaned sample) and after (cleaned sample) the treatment in a cleaning system were studied. The influence of different doses of UV-B radiation on cell shape and motility was also investigated. No statistically significant effects of increasing concentrations of the waste substances both before and after the cleaning process (except inhibition by the undiluted uncleaned effluent) on the growth rate in E. gracilis were observed. Cell shape turned out to be an unreliable physiological parameter for assessing the toxicity at lower concentrations of waste water substances. No significant patterns could be observed in the response of the cell shape to the different concentrations of the waste water substances or to UV-B radiation. Motility has been concluded to be a more sensitive parameter than cell shape. However, no clear patterns were observed in the response of the motility to the different concentrations of the waste water substances studied. Increasing concentrations of the uncleaned sample demonstrated a defense against UV-B radiation, due to the high absorbance in the UV-B range, when effects on motility were examined. We conclude that contrary to the results reported in the literature earlier, cell shape and motility of E. gracilis are not universal physiological parameters for bioassessment at lower concentrations of toxic substances, including metals such as copper and zinc from the pulp and paper industry. The long-duration tests had, in general, higher significance than those of short duration.

Animals↗
PubMed1991

Bacterial motility: handedness and symmetry.

Many bacteria swim by rotating thin helical filaments that extend into the external medium, as with common bacteria, or run beneath the outer membrane, as with spirochetes. Each filament is driven at its base by a motor that turns alternately clockwise and counterclockwise. The motor-filament complex is called a flagellum. Other kinds of bacteria glide, but their organelles of locomotion are not known. Since bacteria are microscopic and live in an aqueous environment, they swim at low Reynolds' number; cyclic motion works (e.g. rotation of a helix) but reciprocal motion does not (e.g. stroking of a singly hinged oar). By measuring concentrations of certain chemicals as they move through their environment, making temporal comparisons and modulating the direction of flagellar rotation, bacteria accumulate in regions that they find more favourable. Studies of bacterial chemotaxis are highly advanced, particularly for the peritrichously flagellated species Escherichia coli. A great deal is known about chemoreception, receptor-flagellar coupling and adaptation. Recently it has been found that E. coli can aggregate in response to signals generated by the cells themselves. Complex patterns form with remarkable symmetries.

Bacterial Physiological Phenomena↗
PubMed1983

Chronobiology at the cellular and molecular levels: models and mechanisms for circadian timekeeping.

This review considers cellular chronobiology and examines, at least in a superficial way, several classes of models and mechanisms that have been proposed for circadian rhythmicity and some of the experimental approaches that have appeared to be most productive. After a brief discussion of temporal organization and the metabolic, epigenetic, and circadian time domains, the general properties of circadian rhythms are enumerated. A survey of independent oscillations in isolated organs, tissues, and cells is followed by a review of selected circadian rhythms in eukaryotic microorganisms, with particular emphasis placed on the rhythm of cell division in the algal flagellate Euglena as a model system illustrating temporal differentiation. In the ensuing section, experimental approaches to circadian clock mechanisms are considered. The dissection of the clock by the use of chemical inhibitors is illustrated for the rhythm of bioluminescence in the marine dinoflagellate Gonyaulax and for the rhythm of photosynthetic capacity in the unicellular green alga Acetabularia. Alternatively, genetic analysis of circadian oscillators is considered in the green alga Chlamydomonas and in the bread mold Neurospora, both of which have yielded clock mutants and mutants having biochemical lesions that exhibit altered clock properties. On the basis of the evidence generated by these experimental approaches, several classes of biochemical and molecular models for circadian clocks have been proposed. These include strictly molecular models, feedback loop (network) models, transcriptional (tape-reading) models, and membrane models; some of their key elements and predictions are discussed. Finally, a number of general unsolved problems at the cellular level are briefly mentioned: cell cycle interfaces, the evolution of circadian rhythmicity, the possibility of multiple cellular oscillators, chronopharmacology and chronotherapy, and cell-cycle clocks in development and aging.

Acetabularia↗
PubMed2005

A twelve-step program for evolving multicellularity and a division of labor.

The volvocine algae provide an unrivalled opportunity to explore details of an evolutionary pathway leading from a unicellular ancestor to multicellular organisms with a division of labor between different cell types. Members of this monophyletic group of green flagellates range in complexity from unicellular Chlamydomonas through a series of extant organisms of intermediate size and complexity to Volvox, a genus of spherical organisms that have thousands of cells and a germ-soma division of labor. It is estimated that these organisms all shared a common ancestor about 50 +/- 20 MYA. Here we outline twelve important ways in which the developmental repertoire of an ancestral unicell similar to modern C. reinhardtii was modified to produce first a small colonial organism like Gonium that was capable of swimming directionally, then a sequence of larger organisms (such as Pandorina, Eudorina and Pleodorina) in which there was an increasing tendency to differentiate two cell types, and eventually Volvox carteri with its complete germ-soma division of labor.

Animals↗
PubMed1993

Structural characterization of the cyanelle peptidoglycan of Cyanophora paradoxa by 252Cf plasma desorption mass spectrometry and fast atom bombardment/tandem mass spectrometry.

A strategy for the structural characterization of the four major NaBH4-reduced peptidoglycan monomers derived from muramidase-digested peptidoglycan from the cyanelles of the flagellate Cyanophora paradoxa Korschikoff is described. Initial molecular weight determination of these glycopeptides was performed by positive and negative ion plasma desorption mass spectrometry. Due to the presence of two pairs of disaccharide tripeptide and disaccharide tetrapeptide monomers differing in mass by 112 units, respectively, an as yet unknown peptidoglycan modification either at the carbohydrate or at the peptide moiety was assumed. beta-Elimination of the disaccharide unit from the unreduced peptidoglycan monomers yielded the corresponding (modified) N1-lactyltripeptides and -tetrapeptides, respectively. These peptides, N-terminally blocked with lactic acid, unambiguously showed the modification to be located on the peptide moiety. By positive ion fast atom bombardment/hybrid tandem mass spectrometry of the reduced peptidoglycan monomers as well as of the corresponding deglycosylated monomers (= N1-lactylpeptides) the modification was determined to be linked to the glutamic acid moiety. Based on combined data from plasma desorption mass spectrometry, tandem mass spectrometry, accurate mass measurement and amino acid analysis of the acid hydrolysate after derivatization with o-phthaldialdehyde by high-performance liquid chromatography we could establish the structure of the modification as N-acetylputrescine. Finally, the confirmation of the linkage of the glutamic acid to diaminopimelic acid via the gamma-COOH was based on the presence of a-type peptide backbone fragment ions in the positive ion plasma desorption mass spectra of the modified N1-lactylpeptides.

Amino Acid Sequence↗
PubMed2002

Transient concentration of a gamma-tubulin-related protein with a pericentrin-related protein in the formation of basal bodies and flagella during the differentiation of Naegleria gruberi.

The distribution of two proteins in Naegleria gruberi, N-gammaTRP (Naegleria gamma-tubulin-related protein) and N-PRP (Naegleria pericentrin-related protein), was examined during the de novo formation of basal bodies and flagella that occurs during the differentiation of N. gruberi. After the initiation of differentiation, N-gammaTRP and N-PRP began to concentrate at the same site within cells. The percentage of cells with a concentrated region of N-gammaTRP and N-PRP was maximal (68%) at 40 min when the synthesis of tubulin had just started but no assembled microtubules were visible. When concentrated tubulin became visible (60 min), the region of concentrated N-gammaTRP and N-PRP was co-localized with the tubulin spot and then flagella began to elongate from the region of concentrated tubulin. When cells had elongated flagella, the concentrated N-gammaTRP and N-PRP were translocated to the opposite end of the flagellated cells and disappeared. The transient concentration of N-gammaTRP coincided with the transient formation of an F-actin spot at which N-gammaTRP and alpha-tubulin mRNA were co-localized. The concentration of N-gammaTRP and formation of the F-actin spot occurred without the formation of microtubules but were inhibited by cytochalasin D. These observations suggest that the regional concentration of N-gammaTRP and N-PRP is mediated by actin filaments and might provide a site of microtubule nucleation for the assembly of newly synthesized tubulins into basal bodies and flagella.

Actins↗
PubMed2003

Gliding movement in Peranema trichophorum is powered by flagellar surface motility.

A colorless euglenoid flagellate Peranema trichophorum shows unique unidirectional gliding cell locomotion on the substratum at velocities up to 30 micro m/s by an as yet unexplained mechanism. In this study, we found that (1) treatment with NiCl(2) inhibited flagellar beating without any effect on gliding movement; (2) water currents applied to a gliding cell from opposite sides caused detachment of the cell body from the substratum. With only the anterior flagellum adhering to the substratum, gliding movement continued along the direction of the anterior flagellum; (3) gentle pipetting induced flagellar severance into various lengths. In these cells, gliding velocity was proportional to the flagellar length; and (4) Polystyrene beads were translocated along the surface of the anterior flagellum. All of these results indicate that a cell surface motility system is present on the anterior flagellum, which is responsible for cell gliding in P. trichophorum.

Animals↗
PubMed1994

Glutamylated tubulin probed in ciliates with the monoclonal antibody GT335.

Microtubular networks are extensively developed in many ciliate species. In several of them, we investigate the occurrence of the post-translational glutamylation of tubulin [Eddé et al., 1990: Science 247:82-85; Eddé et al., 1991: J. Cell. Biochem. 46:134-142] using as a probe for such modified tubulin, the monoclonal antibody GT335 [Wolff et al., 1992: Eur. J. Cell Biol. 59:425-432]. Results obtained in Paramecium strongly suggest that both axonemal and cytoplasmic tubulin are glutamylated. As in the vertebrate brain tubulin so far tested, the GT335 epitope is located at the carboxy-terminal fragment of cytoplasmic tubulin removed by subtilisin treatment. Immunoblotting and immunofluorescence experiments reveal that, unlike tubulin acetylation, glutamylation is not restricted to cold-resistant microtubules. In addition, immunofluorescence studies performed on dividing cells show that glutamylation takes place soon after the polymerization of microtubules. Finally, glutamylated tubulin is also detected in the ciliate species Euplotes, Tetrahymena, and Paraurostyla. Together with results obtained on flagellate species, this suggests that tubulin glutamylation came out early in the course of eukaryotic evolution and has been widely exploited in various cellular strategies.

Animals↗
PubMed1989

Algorithm to estimate cell biovolume using image analyzed microscopy.

This paper describes an algorithm for calculating the biovolume of cells with simple shapes, such as bacteria, flagellates, and simple ciliates, from a 2-dimensional digital image. The method can be adapted to any image analysis system which allows access to the binary cell image--(i.e., the pixels, or (x,y) points, composing the cell. The cell image is rotated to a standard orientation (horizontal), and a solid of revolution is calculated by digital integration. Verification and a critical assessment of the method are presented. The algorithm accounts for irregularities in cell shape that conventional methods based on length, width, and geometrical formulas do not.

Algorithms↗