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[Development of cyst-like cells of the flagellate Leptomonas oncopelti in the midgut of the hemipteran Oncopeltus fasciatus].

The structure of cyst-like cells of Leptomonas oncopelti (Trypanosomatidae) found in the midgut of the bug Oncopeltus fasciatus (Lygaeidae) was examined with light and electron microscopy. The formation of "cysts" begins with an unequal division of active flagellates with promastigote configuration. Cytokinesis starts on the lateral side of the flagellate, and then the cleavage furrow moves toward the apical end of the cell. The anterior part of a smaller daughter cell, referred to as cell C1, remains associated with the flagellum of maternal promastigote. C1 divides twice to give rise first to two equivalent cells (C2), and then to four morphologically similar cells (C3). C2 join with each other, and afterwards C3 attach between themselves as well via short cytoplasmic outgrowths, which appear instead flagella. In the point of outgrowth attachment of only one C2 and then of only one C3 to maternal flagellum zonal desmosomes occur. C1--C3 of L. oncopelti are similar to so-called straphangers (cyst-like parasites attached to the flagellum of maternal promastigote) known in some species of the genera Leptomonas and Blastocrithidia. Basal bodies are present in C1 and C2 but not in C3. DNA fibrils in the kinetoplast lack their common circular configuration, they progressively condense to form a disordered mass. C3 chromatin becomes denser to acquire eventually a characteristic "labyrinthine structure" looking like a huge bundle of whorled filaments 3-5 nm width. Inside this bundle there are channels of 10-12 nm in diameter filled with karyoplasm. On becoming ovoid, C3 are separated from the maternal promastigote flagellum and differentiate into mature "cysts". Straphangers C1--C3 and mature "cysts" lack any visible outer extracellular protective envelope (cyst wall). Instead, these cells have a cortical complex made of a reinforced plasmatic membrane underlined by a layer of a dense granular cytoplasm free of subpellicular microtubules. The mature "cyst" endoplasm shows a high electron density, and because of this identification of the majority of cellular organelles is next to impossible. Nevertheless, in both C3 and mature "cysts" some unusual membranes are seen composed of two electron lucent layers, with a single electron dense layer in between.

Animals↗

Problems in the comparative physiology of some trypanosomatid flagellates.

There is considerable evidence that trypanosomatid species vary in their metabolic characteristics, that they vary in the mechanisms by which they control these characteristics, and that a single species may vary metabolically without varying structurally. Studies to date from a number of laboratories also indicate there is still reason to believe that metabolic characteristics of trypanosomatid flagellates, as manifest in culture, are at least correlated (in some cases) with the behavior of the flagellate species in the metazoan host. It appears that our major tasks for the next several years are 1. to discover the extent to which these correlations are manifestations of characteristics required for life in the metazoan host, 2. to discover the extent to which these correlations are manifestations of characteristics which determine infection site within the metazoan host, 3. to discover which if any metabolic control mechanisms contribute to the multiplicity of clinical infections often seen in this group of protozoa, and 4. to discover the critical links between energy metabolism, the control of that metabolism, and life inside a host or host cell.

Culture Media↗

[Phagocytosis and endocytosis in the colorless flagellate Thaumatomonas lauterborni].

Phago- and endocytosis have been studied in the colourless flagellate T. lauterborni using electron microscope. The coated pits are formed on the dorsal surface of the cells and in the flagellar pocket; then they are transformed into coated vesicles and transported into the ventral part of the cell loosing their clathrin coat. The storing of small vesicles in the ventral groove region is constant. To begin to feed a flagellate stops and produces within several seconds long ramified filopodia from the ventral groove. These filopodia serve to phagocyte bacteria. Small ventral vesicles represent the membrane pull which is necessary for a quick formation of the vast surface of filopodia. By means of peroxidase reaction in was shown that these vesicles were of endocytotic origin, rather than being the product of the Golgi apparatus functioning.

Animals↗

Enteritis in turkeys associated with an unusual flagellated protozoan (Cochlosoma anatis).

Outbreaks of enteritis associated with an unusual flagellated protozoan occurred in six California turkey flocks during the summer of 1992. Certain morphological and ultrastructural details of the parasite, which resembles Cochlosoma anatis, are illustrated with scanning and transmission electron micrographs. The flagellate attached to the intestinal mucosa by means of a sucker-like apparatus, and circular impressions of the sucker were created on the surface epithelium. Histological lesions were characterized by blunting and fusion of villi; cellular infiltration of the lamina propria with lymphocytes, plasma cells, histiocytes, and heterophils; and increased numbers of mitotic figures in crypt epithelium.

Animals↗

[The classification of the morphological forms of flagellates in the family Trypanosomatidae].

The partial revision of the generally accepted classification of the morphological forms of trypanosomatid flagellates (Hoare, Wallace, 1966) is proposed. Only "flagellar" characteristics without evaluation of the form of the body and systematic position of the flagellates are used in the new variant of the classification. Six basic morphological forms of trypanosomatids have been included in proposed scheme: amastigotes, endomastigotes, promastigotes, opisthomastigotes, epimastigotes, trypomastigotes.

Animals↗

The position of flagellated protists in the system of lower eukaryotes.

The division of Eukaryota into 8 kingdoms united into 3 superkingdoms is briefly discussed with the rejection of the hypothesis of symbiogenesis and of primary nature of the dinokaryon. It is asserted that Pascher's main hypothesis remains valid. The heterotrophic flagellated and amoeboid forms must be included into different groups of flagellated beings. Some of them: Saprolegniomycetes, Hyphochytridiomycetes, Slopalinata, Cryptaxohelid Heliozoa have been already put into chromophytan branch. Others differ from autotrophic groups more sharply and their taxonomic position may be stated now only tentatively and will be specified as a result of future studies.

Animals↗

[The cell biology of amebas and ameba-flagellates--parasites of man and animals].

The majority of parasitic amoebae and amoeba-flagellates are facultative parasites of animals and humans and only a few of them are obligate parasites (see reviews: Sopina, 1997; Visvesvara, Stehr-Green, 1990). Among the latter Entamoeba histolytica and among the former Naegleria fowleri and several species of Acanthamoeba are most dangerous for humans. It is still unclear whether Balamuthia mandrillaris, pathogenic for monkeys and humans, may be either obligate or facultative parasite (Visvesvara et al., 1993). Endolimax nana and Iodamoeba butschlii are commensals of humans and some animals. This review is devoted to cell biology of the above genera to provide a better understanding of cell-biological aspects of interrelations between these parasites and their hosts. In the Russian text-books on medical and veterinary parasitology these parasitic protozoa have never been regarded in terms of cell biology, and the available information seems to become out of date. This review is aimed to fill these gaps. Evidence on the ultrastructure of the above parasites, in particular that of their mitotic chromosomes, mechanisms of attachment to substratum of many of these, of locomotion and endocytosis, is provided and discussed in addition to the problem of differentiation of Naegleria amoebae into flagellates.

Amebiasis↗

Description of a Giardia varani-like flagellate from a water monitor, Varanus salvator, from Malaysia.

A Giardia varani Lavier, 1923-like flagellate was found in the feces of a captive water monitor, Varanus salvator, originally caught wild from an unknown location in Malaysia. The parasite is similar in size and shape to Giardia lamblia, except that median bodies are rare and cysts are binucleate. A description of both the trophozoite and cyst stage of this flagellate is provided.

Animals↗

[Parasitic amoebae and amoebo-flagellates from the Lobosea and heterolobosea classes].

Different cited evidences on parasitic amoebae and amoebo-flagellates belonging to the Lobosea and Heterolobosea classes (Pages, 1987) have been reviewed. Special attention is paid to various degree of their adaptation to parasitic mode of life, which ranges from a parasitism on a border with commensalism to true parasitism (both facultative and obligatory ones). Besides the coprophilous and commensal species, the number of true parasites among the Lobosea and Heterolobosea classes is comparatively small. In many cases, both facultative and obligatory parasites cause the death of their hosts. Apparently this strongly pronounced pathogenicity of parasitic amoebae and amoebo-flagellates suggest a recent origin of such parasite-host systems. Pathogens of amoebic dysentry, primary amoebic meningoencephalitis and granulomatous amoebic encephalitis are specially considered. In the Russian text-books on a parasitology the information on most of them, except Entamoeba hystolytica, is either totally absent or very scare and out of date.

Amoeba↗

Purification and characterization of basal apparatuses from a flagellate green alga.

Basal apparatuses consisting of two basal bodies and several attached fibers were isolated from the naked green flagellate Spermatozopsis similis by detergent extraction and mechanical disintegration. Sucrose density centrifugation yielded highly enriched basal apparatuses as shown by electron microscopy. SDS-PAGE revealed the absence of histones, indicating the removal of nuclear contaminations from the isolated basal apparatuses. A mass spectrometric analysis of the carboxyterminal peptides of alpha tubulin documented detyrosination and glutamylation as posttranslational modifications and showed that some 5% of the alpha tubulin carries a polyglutamyl side chain which can reach at least 17 residues in length. Monoclonal antibodies raised against the purified basal apparatuses were used to characterize novel components in the basal apparatus. A 210-kD component identified by mAB BAS (basal apparatus of Spermatozopsis) 1.4 was localized in the flagellar transitional region by immunogold electron microscopy. Antibody BAS 16.4 reacted with two high molecular weight bands (approximately 265 and 240 kD) in Western blotting and decorated a fiber attached to the proximal end of the basal bodies. Immunofluorescence staining of isolated cytoskeletons with these mABs demonstrated that the antigens are also present in the basal apparatuses of Chlamydomonas reinhardtii and Dunahella bioculata. These antibodies are useful tools for the molecular cloning of components from the basal apparatus.

Amino Acid Sequence↗

Use of computer-assisted motion analysis for quantitative measurements of swimming behavior in peritrichously flagellated bacteria.

An assay was developed which identifies individual bacterial tumbles and so allows rapid, quantitative measurements of tumble frequency in free-swimming bacteria. Tumble frequency is modulated by cells to enable chemotaxis. Mutations in the chemotaxis signal transduction pathway typically have phenotypes of altered tumble frequency. The purpose of this assay is to quantitatively measure steady-state tumble frequency to enable comparisons of mutant strain phenotypes. It was developed using Escherichia coli but should be applicable to other species with a peritrichous flagellation pattern, such as Salmonella typhimurium. Tumbles are defined by a combination of the parameters rate of change of direction and swimming speed, with a rapid change of direction defining the beginning of a tumble and increased swimming speed defining the end. These parameters have previously been shown to be correlated with tumbles in general but not used to identify discrete tumble events. The computer assay was validated by comparing its results with manual observations by eye. The assay was intended to be most sensitive to swimming patterns similar to wild type so as to resolve subtle changes which would result from partial-function mutations. It quantitatively detects extreme behavioral phenotypes as well and can be modified to increase resolution at either extreme if necessary.

Chemotaxis↗

Independent mechanisms are utilized for the coordinate and transient accumulation of two differentiation-specific mRNAs during differentiation of Naegleria gruberi amoebae into flagellates.

During the differentiation of Naegleria gruberi amoebae into flagellates, four differentiation-specific (DS) mRNAs are transiently and coordinately accumulated. Three of the four DS mRNAs, Class II, III, and IV, encode alpha-tubulin, beta-tubulin, and flagellar calmodulin, respectively. The protein product of the Class I mRNA has not been identified. We examined the effects of inhibition of protein synthesis on transcription and accumulation of beta-tubulin mRNA and Class I mRNA to understand the mechanism of coordinate regulation. Inhibition of protein synthesis at the beginning of differentiation completely blocked transcription of the beta-tubulin gene. Addition of cycloheximide at 30 or 40 min after initiation of differentiation inactivated transcription of the beta-tubulin gene in less than 10 min as judged by nuclear run-on experiments. However, once differentiation had proceeded for more than 50 min, inhibition of protein synthesis did not inactivate transcription of beta-tubulin mRNA was more active in cycloheximide-treated cells than in control cells. Cycloheximide treatment at the initiation of the differentiation also blocked transcription of the Class I gene. However, addition of the drug after 30 min had no significant effect on the transcription of the Class I gene. Cycloheximide treatment also increased the half-lives of beta-tubulin and Class I mRNA drastically. These data suggest that: (1) the transient accumulation of the two DS mRNAs during differentiation are regulated by changing both the rate of transcription and the stability of the mRNAs; (2) protein synthesis is required for the transcriptional and post-transcriptional regulations; (3) the transcriptional regulation mechanisms of the beta-tubulin gene and that of the Class I gene are distinct; and (4) the transcription of the beta-tubulin gene is regulated by different mechanisms during differentiation.

Animals↗

Structure and expression of a gene encoding the large subunit of ribulose-1,5-bisphosphate carboxylase (rbcL) in the colourless euglenoid flagellate Astasia longa.

A gene encoding the large subunit of ribulose-1,5-bisphosphate carboxylase (Rubisco) was identified on a circular 73 kb DNA from the colourless euglenoid flagellate Astasia longa. The rbcL gene of Astasia extends over 3968 bp. It is a split gene interrupted by seven introns as compared to nine intervening sequences in the rbcL gene of the phylogenetically related Euglena gracilis. Coding sequences as well as the positions of the introns within this gene are highly conserved in comparison with the Euglena rbcL except that two introns are missing in Astasia. The alignment of the amino acid sequences deduced from the nucleotide sequences of rbcL of Astasia and Euglena shows 82% identical amino acids whereas 15% of the amino acids represent conservative changes. A 1.5 kb transcript of the rbcL gene was revealed by northern blot analysis of Astasia RNA. By immunoblot analysis the gene product of rbcL was detected as a 53 kDa polypeptide. Genes for components of the chloroplast transcriptional and translational systems encoded by chloroplast DNA of plants and green algae are conserved on the 73 kb DNA of Astasia [24, 25, 26]. From our finding that Astasia obviously is capable of synthesizing the Rubisco large subunit one must conclude that these genes are expressed and form functional plastid transcriptional and translational systems.

Amino Acid Sequence↗

Responses of the photosynthetic flagellate, Euglena gracilis, to hypergravity.

Motility and orientation has been studied in the unicellular photosynthetic flagellate, Euglena gracilis, using real time image analysis capable of tracking up to 200 cells simultaneously in the slow rotating centrifuge microscope (NIZEMI) which allows one to observe the cells' swimming behavior during centrifugation accelerations between 1 g and 5 g. At 1 g the cells show a weak negative gravitaxis, which increases significantly at higher accelerations up to about 3 g. Though most cells were capable of swimming even against an acceleration of 4.5 g, the degree of gravitaxis decreased and some of the cells were passively moved downward by the acceleration force; this is true for most cells at 5 g. The velocity of cells swimming against 1 g is about 10% lower than that of cells swimming in other directions. The velocity decreases even more drastically in cells swimming against higher acceleration forces than those at 1 g. The degree of gravitactic orientation drastically decreases after short exposure to artificial UV radiation which indicates that gravitaxis may be due to an active physiological perception rather than a physical effect such as an asymmetry of the center of gravity within the cell.

Acceleration↗

The role of cAMP in flagellation of Salmonella typhimurium.

A mutational alteration either in adenylate cyclase (cya-) or in cyclic-3'5'-AMP (cAMP) receptor protein (crp-) rendered Salmonella typhimurium incapable of producing flagella. The amount of mRNA specific for flagellin in these mutants was almost negligible when assayed in an in vitro protein synthesizing system. A secondary mutation cfs, partially suppressing the cya- mutation, was identified among the revertants of cya-. A mutation in the same cistron as cfs resulted in a non-flagellate phenotype either by itself or in combination with cfs. The cistron, which was given the gene symbol flaT, was located between flaE and flaL. It was suggested that cAMP receptor protein together with cAMP modulates the gene flaT, which in turn acts as a positive effector on the synthesis of active mRNA specific for flagellin.

Adenylyl Cyclases↗

Organization and nucleotide sequence of ribosomal RNA genes on a circular 73 kbp DNA from the colourless flagellate Astasia longa.

Three tandemly arranged repeats (A, B, C) of 16S and 23S rDNA, and one supplementary (S) 16S rDNA adjacent to the 16S rDNA of repeat A, are present within an 18 kbp segment of a circular 73 kbp DNA from the colourless flagellate Astasia longa. The repeat units are separated by a short region containing a 5S rRNA gene and a gene for tRNA-Val (UAC). Sequence comparisons reveal 78%, 81%, and 67% identical nucleotides of the 23S rDNA (A), the 16S rDNA (B), and the 5S rDNA (A), respectively, with the corresponding genes of the Euglena gracilis chloroplast genome. As in Euglena chloroplasts, the 3'-terminal portion of the 23S rDNA is homologous to the 4.5S rRNA gene of higher plant chloroplast genomes. These results are supportive of a common evolutionary origin for the Astasia 73 kbp DNA and the Euglena 145 kbp chloroplast DNA.

Animals↗

Micromorphology of Gram-negative hydrogen bacteria. I. Cell morphology and flagellation.

The cell morphology, the arrangement and fine structure of flagella and the piliation of the following Gram-negative aerobic hydrogen bacteria have been studied: Alcaligenes eutrophus, Alcaligenes paradoxus, Alcaligenes ruhlandii, Pseudomonas flava, Pseudomonas pseudoflava, Pseudomonas palleronii, Pseudomonas facilis, Aquaspirillum autotrophicum, Paracoccus denitrificans, Corynebacterium autotrophicum, and strains MA 2 and SA 35. The identity of the bacteria was examined by their substrate spectra and type of flagellation. Three types of flagellar fine structure were differentiated. The presence of pili was noted in strains of Alcaligenes paradoxus, Pseudomonas flava, P.pseudoflava, P.palleronii, and P.facilis.

Alcaligenes↗

Polarotaxis, gravitaxis and vertical phototaxis in the green flagellate, Euglena gracilis.

A fully automatic computer-controlled video analysis system has been used to study the movement of the green unicellular flagellate, Euglena gracilis in a horizontal or vertical cuvette. In darkness, in the absence of gaseous gradients, most cells swim straight upwards. While in a horizontal cuvette the transition between positive and negative phototaxis is found at about 1.5 W m-2, an excess of 30 W m-2 is required to reverse the upward swimming (due to the combined stimulus of negative gravitaxis and positive phototaxis) in a vertical cuvette. By studying the swimming direction in horizontal and vertical cuvettes in polarized light irradiated from above or from the side, respectively, the dichroic orientation of the photoreceptor molecules can be determined in three dimensions with respect to the axes of the cell; In a horizontal cuvette, in a linearly polarized beam from above, the cells orient predominantly at an angle of about 30 degrees clockwise off the electric dipole transition moment as seen from above. The behavior in a vertical cuvette with polarized light entering from above indicates that the photoreceptor pigments are dichroically oriented 60 degrees counterclockwise from the flagellar plane (seen from the front end of the cell). Experiments with horizontal polarized light indicate that the photoreceptor transition moment deviates 25 degrees clockwise off the long axis of the cell.

Animals↗