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M Melkonian

Publications and source records attributed to M Melkonian.

48 records · Page 3Linked to original sources
PubMed1987

Basal body reorientation mediated by a Ca2+-modulated contractile protein.

A rapid, Ca2+-dependent change in the angle between basal bodies (up to 180 degrees) is associated with light-induced reversal of swimming direction (the "photophobic" response) in a number of flagellated green algae. In isolated, detergent-extracted, reactivated flagellar apparatus complexes of Spermatozopsis similis, axonemal beat form conversion to the symmetrical/undulating flagellar pattern and basal body reorientation (from the antiparallel to the parallel configuration) are simultaneously induced at greater than or equal to 10(-7) M Ca2+. Basal body reorientation, however, is independent of flagellar beating since it is induced at greater than or equal to 10(-7) M Ca2+ when flagellar beating is inhibited (i.e., in the presence of 1 microM orthovanadate in reactivation solutions; in the absence of ATP or dithiothreitol in isolation and reactivation solutions), or when axonemes are mechanically removed from flagellar apparatuses. Although frequent axonemal beat form reversals were induced by varying the Ca2+ concentration, antiparallel basal body configuration could not be restored in isolated flagellar apparatuses. Observations of the photophobic response in vivo indicate that even though the flagella resume the asymmetric, breaststroke beat form 1-2 s after photostimulation, antiparallel basal body configuration is not restored until a few minutes later. Using an antibody generated against the 20-kD Ca2+-modulated contractile protein of striated flagellar roots of Tetraselmis striata (Salisbury, J. L., A. Baron, B. Surek, and M. Melkonian, 1984, J. Cell Biol., 99:962-970), we have found the distal connecting fiber of Spermatozopsis similis to be immunoreactive by indirect immunofluorescence and immunogold electron microscopy. Electrophoretic and immunoblot analysis indicates that the antigen of S. similis flagellar apparatuses consists, like the Tetraselmis protein, of two acidic isoforms of 20 kD. We conclude that the distal basal body connecting fiber is a contractile organelle and reorients basal bodies during the photophobic response in certain flagellated green algae.

Calcium↗
PubMed1984

Striated flagellar roots: isolation and partial characterization of a calcium-modulated contractile organelle.

We report the isolation of striated flagellar roots from the Prasinophycean green alga Tetraselmis striata using sedimentation in gradients of sucrose and flotation on gradients of colloidal silica. PAGE in the presence of 0.1% SDS demonstrates that striated flagellar roots are composed of a number of polypeptides, the most predominant one being a protein of 20,000 Mr. The 20,000 Mr protein band represents approximately 63% of the Coomassie Brilliant Blue staining of gels of isolated flagellar roots. Two-dimensional gel electrophoresis (isoelectric focusing and SDS PAGE) resolves the major 20,000 Mr flagellar root protein into two components of nearly identical Mr, but of differing isoelectric points (i.e., pl's of 4.9 and 4.8), which we have designated 20,000-Mr-alpha and 20,000-Mr-beta, respectively. Densitometric scans of two-dimensional gels of cell extracts indicate that the 20,000-Mr-alpha and -beta polypeptides vary, in their stoichiometry, between 2:1 and 1:1. This variability appears to be related to the state of contraction or extension of the striated flagellar roots at the time of cell lysis. Incubation of cells with 32PO4 followed by analysis of cell extracts by two-dimensional gel electrophoresis and autoradiography reveals that the more acidic 20,000-Mr-beta component is phosphorylated and the 20,000-Mr-alpha component contains no detectable label. These results suggest that the 20,000-Mr-alpha component is converted to the more acidic 20,000-Mr-beta form by phosphorylation. Both the 20,000-Mr-alpha and -beta flagellar root components exhibit a calcium-induced reduction in relative electrophoretic mobilities in two-dimensional alkaline urea gels. Antiserum raised in rabbits against the 20,000-Mr protein binds to both the 20,000-Mr-alpha and 20,000-Mr-beta forms of the flagellar root protein when analyzed by electrophoretic immunoblot techniques. Indirect immunofluorescence on vegetative or interphase cells demonstrate that the antibodies bind to two cyclindrical organelles located in the anterior region of the cell. Immunocytochemical investigations at ultrastructural resolution using this antiserum and a colloidal gold-conjugated antirabbit-IgG reveals immunospecific labeling of striated flagellar roots and their extensions. We conclude that striated flagellar roots are simple ion-sensitive contractile organelles composed predominantly of a 20,000 Mr calcium-binding phosphoprotein, and that this protein is largely responsible for the motile behavior of these organelles.

Calcium↗
PubMed1982

The functional analysis of the flagellar apparatus in green algae.

The flagellar apparatus of green algae is a complex organelle of great structural diversity. Diversity exists with respect to numbers of flagella, flagellar appendages (e.g., scales), origin of flagella in relation to cell symmetry, basal body and transitional region ultrastructure, fibrous structures interconnecting basal bodies and flagellar root systems. The flagellar apparatus performs several different functions in a single cell and also probably functions differently in different green algal genera. In addition to being an organelle which moves the cell through an aqueous environment the flagellar apparatus reacts to the mechanical, chemical, gravitational and light stimulation of the cell. It also plays an important role during sexual fusion of isogamous, anisogamous and oogamous species of green algae. Furthermore it contains several microtubule-organising centres (MTOCs) and in most flagellated green algae these MTOCs organise the whole microtubular system of the cell. In this review the different strategies which are currently employed to evaluate the functional significance of various structures of the flagellar apparatus in green algae are discussed. Emphasis is given to those structures which are not directly engaged in generating the motive force of flagella.

Cell Membrane↗
PubMed1981

Comparative ultrastructure of eyespot membranes in gametes and zoospores of the green alga Ulva lactuca (Ulvales).

Eyespot membranes in zoospores, and male and female gametes of the green alga Ulva lactuca, were studied comparatively by the freeze-fracture technique. The plasmalemma and the outer chloroplast envelope membrane overlying the eyespot lipid globules are specialized in all 3 types of reproductive cells. In the eyespot region the protoplasmic face (PF) of the outer chloroplast envelope membrane contains significantly more intramembraneous particles (IMP) compared to membrane areas outside the eyespot: in female gametes there are 2.5 times more IMP/micrometers 2, in zoospores 3 and in male gametes about 4. Small size-class IMP (4--6 nm diameter) are particularly abundant on both fracture faces of the outer chloroplast membrane, but size-class distribution is not significantly different between membrane areas inside and outside the eyespot region. The total number of IMP/eyespot on the PF of the outer chloroplast membrane was calculated to be 4900 in male gametes, 5500 in female gametes and 11 200 in zoospores. The results are discussed in accordance with the view that these membrane specializations participate in photoreception relating to green algal phototaxis. Evidence is presented that there is a correlation between IMP numbers per eyespot in the outer chloroplast envelope membrane and the different phototactic behaviour of gametes compared to zoospores in Ulva.

Chlorophyta↗
PubMed1981

Sterol-deficient domains correlate with intramembrane particle arrays in the plasma membrane of Chlamydomonas reinhardii.

The planar distribution of 3-beta-hydroxysterols in the plasmalemma of the green flagellate Chlamydomonas reinhardii has been studied with the freeze-fracture technique using the polyene antibiotic filipin and the saponin tomatin as cytochemical markers. Filipin-sterol complexes were predominantly observed as 25 to 30 nm protuberances on the E-face of the plasmalemma with corresponding invaginations on the P-face. Generally filipin-sterol and tomatin-sterol complexes were randomly distributed, but were virtually absent from sites of intramembrane particle arrays (IMP-arrays, i.e. the flagellar necklace, the flagellar bracelet including strut arrays, the eyespot membrane). The results suggest that IMP-arrays of Chlamydomonas are deficient in 3-beta-hydroxysterols and may therefore be regarded as special lipid arrays. IMP-arrays might require an altered lipid environment for proper function. Some functional aspects of IMP-arrays in Chlamydomonas are discussed.

Cell Membrane↗
PubMed1980

Ultrastructural aspects of basal body associated fibrous structures in green algae: a critical review.

Ultrastructural aspects of fibrous structures associated with basal bodies of green algae are critically discussed. It is apparent that variation among these structures is much greater than in microtubular flagellar root systems and it is therefore suggested that fibrous structures may be more useful than microtubular roots in elucidating phylogenetic relationships within the Chlorophyceae sensu Stewart and Mattox and the Prasinophyceae sensu Christensen. Two main types of fibrous structures are distinguished: (1) Connecting fibres (these connect different basal bodies); (2) Fibrous roots (these originate at basal bodies and terminate somewhere else in the cell). Fibrous roots are of two types: (a) microtubular-root associated striated fibres (striation pattern 25-35 nm; system I-fibres); (b) striated fibres composed of a bundle of filaments (filament diameter: 5-10 nm; striation pattern greater than 80 nm; system II fibres). Numbers, disposition and substructure of connecting fibres and fibrious roots are variable in different genera of green algae. In the experimental secretion new observations on fibrous roots in the ulvalean genus Enteromorpha as well as preliminary information on fibrous structures in Carteria obtusa and Bryopsis lyngbyei are included. Functional and evolutionary aspects of fibrous structures associated with the flagellar apparatus of green algae are discussed.

Chlorophyta↗
PubMed1980

Flagellar roots, mating structure and gametic fusion in the green alga Ulva lactuca (Ulvales).

The slightly anisogamous gametes of Ulva lactuca exhibit a cruciate flagellar root system consisting of 4 microtubular roots (4-2-4-2 system) and an elaborate system of fibrous roots associated with the 2-stranded microtubular roots. Two fibres (32-nm striation periodicity; system I fibres) closely underlie each of the 2-stranded roots, while different fibres (150-nm striation periodicity; system II fibres) run parallel to the root microtubules, and are 150-200 nm more internally located. Female gametes have 4 system II fibres, 3 of which are combined into a compound fibre associated with one microtubular root, while the fourth fibre is associated with the opposite root. In male gametes only 2 system II fibres are present, each underlying one of the two 2-stranded roots. A special region of the plasmalemma of both gamete types about 0.5 mum away from the basal bodies and located between 2 adjacent microtubular roots is structurally specialized and acts as a mating structure in gametic fusion. The region is oval-shaped and up to I.I mum long with a maximum diameter of 0.7 mum. A continuous electron-dense boundary layer underlies the plasmalemma at the edges of the mating structure. In both gamete types the mating structure consists of a fuzzy layer of material underlying the plasmalemma and special granules (60 nm diameter) are associated with this layer on its cytoplasmic side. In addition diffuse material overlies the mating structure, especially in male gametes. The mating structure is connected to 3 different kinds of flagellar roots: the boundary layer is linked to a 2-stranded microtubular root and its associated system I fibre; the fuzzy layer of the mating structure is connected with a system II fibre; and in female gametes this is the compound system II fibre. The ultrastructural changes which occur after mixing the 2 gamete types have been followed. Mating structure activation involves contraction of system II fibres (change of striation periodicity to 100 nm), detachment of special granules from the fuzzy layer of the mating structure and their replacement by electron-transparent vesicles at the prospective cell fusion site. Furthermore, release of electron-dense contents from Golgi-derived vesicles in the anterior part of both gamete types precedes cell fusion. Cell fusion is exclusively initiated in a region delimited by the 2 mating structures. After partial dissolution the 2 plasma membranes unite within the mating structure regions. The ultrastructure of gametic fusion in Ulva lactuca is compared to that of other green algae and the significance of flagellar roots in the mating process of green algae is discussed.

Chlorophyta↗
PubMedDate not supplied

Molecular evolutionary analyses of nuclear-encoded small subunit ribosomal RNA identify an independent rhizopod lineage containing the Euglyphina and the Chlorarachniophyta.

The Rhizopoda comprise a diverse assemblage of protists which depend on lobose or filose pseudopodia for locomotion. The biochemical and morphological diversity of rhizopods has led to an uncertain taxonomy. Ribosomal RNA sequence comparisons offer a measure of evolutionary relatedness that is independent of morphology and has been used to demonstrate a polyphyletic origin of the Lobosea. We sequenced complete small subunit ribosomal RNA coding regions from the filose amoebae, Euglypha rotunda and Paulinella chromatophora (Euglyphina) to position these taxa in the eukaryote phylogeny. The neighbor-joining analyses show that E. rotunda and P. chromatophora share a monophyletic origin and are not closely related to any lobose amoebae in our analyses. Instead, the Euglyphina form a robust sister group to the Chlorarachniophyta. These results provide further evidence for the polyphyly of the Rhizopoda and support the creation of a new amoeboid lineage which includes the Euglyphina and the chlorarachniophyte algae; taxa with tubular mitochondrial cristae and filose or reticulate pseudopodia.

Animals↗
PubMedDate not supplied

[Lipid peroxidation processes and the dynamics of shifts in the cholesterol level in plasma and erythrocytes under the influence of low-frequency acoustic oscillations].

Distinct alterations in lipid peroxidation, in content of alpha-tocopherol and cholesterol fractions were observed in rat blood plasma and erythrocyte membranes after long-term low-frequency acoustic oscillations (105 decibels, diurnally, within 3 months). Under these conditions activities of superoxide dismutase and erythrocyte Na+, K+-ATPase were also altered. The rate and direction of the enzymatic activity alterations correlated with the duration of the acoustic oscillation exposure and the patterns studied. Enrichment of blood plasma and erythrocyte membranes with alpha-tocopherol within 2-4 weeks of the acoustic oscillation is considered as an important phenomenon.

Animals↗