scholarly journals Calcium control of waveform in isolated flagellar axonemes of chlamydomonas

1980 ◽  
Vol 86 (2) ◽  
pp. 446-455 ◽  
Author(s):  
M Bessen ◽  
RB Fay ◽  
GB Witman
Keyword(s):  
Basal Bodies ◽  
Membrane Matrix ◽  
Flagellar Membrane ◽  
Matrix Fraction ◽  
Almost All

The effect of Ca(++) on the waveform of reactivated, isolated axonemes of chlamydomonas flagella was investigated. Flagella were detached and isolated by the dibucaine procedure and demembranated by treatment with the detergent Nonidet; the resulting axomenes lack the flagellar membrane and basal bodies. In Ca(++)-buffered reactivation solutions containing 10(-6) M or less free Ca(++), the axonemes beat with a highly asymmetrical, predominantly planar waveform that closely resembled that of in situ flagella of forward swimming cells. In solutions containing 10(-4) M Ca(++), the axonemes beat with a symmetrical waveform that was very similar to that of in situ flagella during backward swimming. In 10(-5) M Ca(++), the axonemes were predominantly quiescent, a state that appears to be closely associated with changes in axomenal waveform or direction of beat in many organisms. Experiments in which the concentrations of free Ca(++), not CaATP(--) complex were independently varied suggested that free Ca(++), not CaATP(--), was responsible for the observed changes. Analysis of the flagellar ATPases associated with the isolated axonemes and the nonidet- soluble membrane-matrix fraction obtained during preparation of the axonemes showed that the axonemes lacked the 3.0S Ca(++)-activated ATPase, almost all of which was recovered in the membrane-matrix fraction. These results indicate that free Ca(++) binds directly to an axonemal component to alter flagellar waveform, and that neither the 3.0S CaATPase nor the basal bodies are directly involved in this change.

Identification of proteins associated with microtubule-organizing centres and filaments in the oral apparatus of the ciliate Paramecium tetraurelia

1990 ◽  
Vol 97 (3) ◽  
pp. 553-563
Author(s):  
GUY KERYER ◽  
FRANCINE IFTODE ◽  
MICHEL BORNENS
Keyword(s):  
Monoclonal Antibody ◽  
Spatial Organization ◽  
Buccal Cavity ◽  
Ultrastructural Localization ◽  
Basal Bodies ◽  
Paramecium Tetraurelia ◽  
Single Polypeptide ◽  
Almost All ◽  
Structure And Activity

In an effort to study the assembly of microtubules in ciliates we have isolated the oral apparatus of Paramecium tetraurelia, a major microtubuie-organizing centre (MTOC) and identified several proteins localized in the fibrillar system associated with basal bodies. Using a monoclonal antibody raised against human centrosomes (CTR210) and a polyclonal antibody (OF1) directed against a 87x103Mr protein of the oral apparatus of Tetrahymena, we observed the same decoration of the oral apparatus of Paramecium, both in situ and after isolation. Ultrastructural localization further showed the presence of the antigens in the fibrillar network that forms a layer under almost all the buccal cavity in close apposition to the basal bodies. CTR210 recognized two sets of polypeptides of Mr72 and 80x103, whereas OF1 recognized a single polypeptide of Mr87x103. Only the 80x103Mr polypeptide was also decorated with the monoclonal antibody MPM-2, previously shown to decorate the oral apparatus of Paramecium and known to react with phosphorylated epitopes in a large variety of MTOCs. All the proteins identified with the three antibodies are insoluble at high ionic strength, display several isoforms and apparently belong to the same fibrillar material. The function of this material in the spatial organization, the structure, and activity of the MTOCs is discussed.

scholarly journals Diversity of Biodeteriorative Bacterial and Fungal Consortia in Winter and Summer on Historical Sandstone of the Northern Pergola, Museum of King John III’s Palace at Wilanow, Poland

2021 ◽  
Vol 11 (2) ◽  
pp. 620
Author(s):  
Magdalena Dyda ◽  
Agnieszka Laudy ◽  
Przemyslaw Decewicz ◽  
Krzysztof Romaniuk ◽  
Martyna Ciezkowska ◽  
...  
Keyword(s):  
Seasonal Changes ◽  
Microbial Community Composition ◽  
Fungal Communities ◽  
Bacterial Strains ◽  
Microbial Biodiversity ◽  
Acidophilic Bacteria ◽  
Almost All ◽  
Generation Sequencing ◽  
King John

The aim of the presented investigation was to describe seasonal changes of microbial community composition in situ in different biocenoses on historical sandstone of the Northern Pergola in the Museum of King John III’s Palace at Wilanow (Poland). The microbial biodiversity was analyzed by the application of Illumina-based next-generation sequencing methods. The metabarcoding analysis allowed for detecting lichenized fungi taxa with the clear domination of two genera: Lecania and Rhinocladiella. It was also observed that, during winter, the richness of fungal communities increased in the biocenoses dominated by lichens and mosses. The metabarcoding analysis showed 34 bacterial genera, with a clear domination of Sphingomonas spp. across almost all biocenoses. Acidophilic bacteria from Acidobacteriaceae and Acetobacteraceae families were also identified, and the results showed that a significant number of bacterial strains isolated during the summer displayed the ability to acidification in contrast to strains isolated in winter, when a large number of isolates displayed alkalizing activity. Other bacteria capable of nitrogen fixation and hydrocarbon utilization (including aromatic hydrocarbons) as well as halophilic microorganisms were also found. The diversity of organisms in the biofilm ensures its stability throughout the year despite the differences recorded between winter and summer.

The Membrane Skeleton of Pseudomicrothorax

1991 ◽  
Vol 100 (4) ◽  
pp. 707-715 ◽  
Author(s):  
IRM HUTTENLAUCH ◽  
ROBERT K. PECK
Keyword(s):  
Spatial Organization ◽  
Membrane Skeleton ◽  
Basal Bodies ◽  
Structural Elements ◽  
Or Groups ◽  
Positive Immunoreactivity ◽  
Cortical Membranes ◽  
Cytoskeletal Elements

The membrane skeleton, or epiplasm, is part of the structurally complex ciliate cortex. It is thought to have skeletal functions concerning the spatial organization of cortical elements such as the basal bodies. Here we report the biochemical and immunological characterization of some components of the purified epiplasm of Pseudomicrothorax dubius. The epiplasm proteins consist of two quantitatively major groups of proteins, one of 76–80x103Mr, the other of 11–13x103Mr, which appear to be the principal structural elements of the epiplasm, and a series of minor components of 62–18x103Mr. Based upon lectin labeling and glycosidase treatment, some of the latter have been identified as glycoproteins. Using affinity-purified antibodies specific for individual glycoproteins or groups of glycoproteins, we were able to localize them in situ by immunoelectron microscopical methods. This in situ localization demonstrates that the glycosylated epitopes, unlike the glycoresidues of membrane proteins, are distributed throughout the entire epiplasmic layer rather than being restricted to regions adjacent to the cortical membranes. Thus, these proteins represent glycosylated, cytoskeletal elements. At least one of these glycoproteins (Mr 62x103) shows positive immunoreactivity with a monoclonal antibody (Pruss anti-IFA) recognizing most intermediate filament (IF) proteins, indicating that IF proteins might be present in protozoan cytoskeletons.

scholarly journals Genotypic diversity of S. mutans in dental biofilm formed in situ under sugar stress exposure

2007 ◽  
Vol 18 (3) ◽  
pp. 185-191 ◽  
Author(s):  
Rodrigo Alex Arthur ◽  
Cínthia Pereira Machado Tabchoury ◽  
Renata de Oliveira Mattos-Graner ◽  
Altair A. Del Bel Cury ◽  
Adriana Franco Paes Leme ◽  
...  
Keyword(s):  
Genotypic Diversity ◽  
Negative Control ◽  
Stress Exposure ◽  
Chain Reaction ◽  
Dental Biofilm ◽  
Polymerase Chain ◽  
Fermentable Carbohydrates ◽  
Almost All ◽  
Frequent Exposure

In situ dental biofilm composition under sugar exposure is well known, but sugar effect on the genotypic diversity of S. mutans in dental biofilm has not been explored. This study evaluated S. mutans genotypic diversity in dental biofilm formed in situ under frequent exposure to sucrose and its monosaccharide constituents (glucose and fructose). Saliva of 7 volunteers was collected for isolation of S. mutans and the same volunteers wore intraoral palatal appliances, containing enamel slabs, which were submitted to the following treatments: distilled and deionized water (negative control), 10% glucose + 10% fructose (fermentable carbohydrates) solution or 20% sucrose (fermentable and EPS inductor) solution, 8x/day. After 3, 7 and 14 days, the biofilms were colleted and S. mutans colonies were isolated. Arbitrarily primed polymerase chain reaction (AP-PCR) of S. mutans showed that salivary genotypes were also detected in almost all biofilm samples, independently of the treatment, and seemed to reflect those genotypes present at higher proportion in biofilms. In addition to the salivary genotypes, others were found in biofilms but in lower proportions and were distinct among treatment. The data suggest that the in situ model seems to be useful to evaluate genotypic diversity of S. mutans, but, under the tested conditions, it was not possible to clearly show that specific genotypes were selected in the biofilm due to the stress induced by sucrose metabolism or simple fermentation of its monosaccharides.

scholarly journals Comparative molecular cytogenetics in Melipona Illiger species (Hymenoptera, Apidae)

Sociobiology ◽  
2018 ◽  
Vol 65 (4) ◽  
pp. 696 ◽  
Author(s):  
Vanderly Andrade-Souza ◽  
Olivia Maria Pereira Duarte ◽  
Cinthia Caroline Cardoso Martins ◽  
Igor Silva Santos ◽  
Márcio Gilberto Cardoso Costa ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Chromosome Number ◽  
Fluorescent In Situ Hybridization ◽  
Molecular Cytogenetics ◽  
Fluorochrome Staining ◽  
Rdna Sites ◽  
Cytogenetic Studies ◽  
Melipona Species ◽  
Almost All

Cytogenetic studies in Melipona are scarce with only 24 species analyzed cytogenetically. Of these, six species had the rDNA sites physically mapped and characterized by Fluorescent in situ Hybridization (fish). The aim of this study was to perform karyotype analyzes on Melipona species from different regions of Brazil, with a greater sampling representative of the Amazonian fauna and using conventional, fluorochrome staining and FISH with heterologous rDNA probes. The predominant chromosome number was 2n = 18, however, the subspecies M. seminigra abunensis and M. s. pernigra showed 2n = 22 chromosomes. The karyotypes were symmetrical, however M. bicolor, M. quadrifasciata, M. flavolineata, M. fuscopilosa, M. nebulosa presented the first pair heteromorphic in length. CMA3+ blocks also exhibited heteromorphism of size and in almost all cases coincided with rDNA sites, except for M. crinita and M. nebulosa, which presented additional non-coincident CMA3+ blocks. The CMA/ rDNA sites were terminal and interstitial in species with high heterochromatic content, and pericentromeric in those species with low heterochromatic content. In addition to pointing out cytogenetic features of cytotaxonomic importance, the reorganization of the genome in Melipona is discussed.

Colonization of depopulated crinoids by symbionts: who comes from the bottom and who from the water column?

2015 ◽  
Vol 95 (8) ◽  
pp. 1607-1612 ◽  
Author(s):  
E.S. Mekhova ◽  
P.Y. Dgebuadze ◽  
V.N. Mikheev ◽  
T.A. Britayev
Keyword(s):  
Water Column ◽  
Field Experiments ◽  
Dispersal Ability ◽  
Long Distance ◽  
Sandy Substrate ◽  
Limited Dispersal ◽  
Set Up ◽  
Almost All ◽  
High Degree

Previous experiments with the comatulid Himerometra robustipinna (Carpenter, 1881) demonstrated intensive host-to-host migration processes for almost all symbiotic species both within host aggregations and among hosts separated by several metres. The aim of this study was to check the ability of symbionts to complete long-distance migrations, by means of two in situ experiments which depopulated the crinoid host. Two different sets of field experiments were set up: exposure of depopulated crinoids (set 1) on stony ‘islands’ isolated from native crinoid assemblages by sandy substrate, and (set 2) in cages suspended in the water column. Hosts from set 1 were exposed for 1, 2, 3 and 4 weeks to assess whether substrate has an influence on the symbionts' long-distance migrations. In set 2 cages were exposed for 10–11 days, aiming to check whether symbionts were able to disperse through the water column with currents. These experiments allow the conclusion that post-settled symbionts can actively migrate among their hosts. Symbionts are able to reach their hosts by employing two different ‘transport corridors’, by drifting or swimming in water column, and by moving on the bottom. Comparison of experimental results allows the division of symbionts into two conventional groups according to the dispersal ability of their post-settled stages: (1) species able to complete long-distance migrations, (2) species unable to migrate or having limited dispersal ability. The finding of the free-living shrimp Periclimenes diversipes Kemp, 1922 in set 2 raises the question about the factors that affect such a high degree of specialization of crinoid assemblages.

Localization of actin in the Tetrahymena basal body-cage complex

1992 ◽  
Vol 103 (3) ◽  
pp. 629-641 ◽  
Author(s):  
J.G. Hoey ◽  
R.H. Gavin
Keyword(s):  
Basal Body ◽  
Specific Binding ◽  
Ultrastructural Level ◽  
Basal Bodies ◽  
Muscle Actin ◽  
Cage Complex ◽  
Contractile System ◽  
Chicken Muscle ◽  
Filament Bundles

In the ciliate cytoskeleton, basal bodies are contained within separate, filamentous cages which are closely associated with basal body microtubules. We have used two polyclonal anti-actin antibodies to localize actin within the basal body-cage complex of Tetrahymena. An antiserum against a Tetrahymena oral apparatus fraction enriched for basal body proteins was produced in rabbits. Agarose-linked chicken muscle actin was used to affinity-purify anti-Tetrahymena actin antibodies from the anti-oral apparatus antiserum. Agarose-linked chicken muscle actin was used to affinity-purify anti-chicken muscle actin antibodies from a commercially available antiserum against chicken muscle actin. Both affinity-purified antibodies were monospecific for Tetrahymena actin on immunoblots containing total oral apparatus protein. The anti-actin antibodies were localized to both somatic and oral basal bodies in Tetrahymena by immunofluorescence microscopy. At the ultrastructural level with the immunogold technique, these antibodies labeled actin epitopes in four distinct regions of the basal body-cage complex: (a) basal body walls, (b) basal plate filaments, (c) proximal-end filaments and (d) cage wall filaments. In addition, the antibody labeled filament bundles that interconnect groups of basal bodies (membranelles) within the oral apparatus. Identical labeling patterns were observed with basal bodies in the isolated oral apparatus, basal bodies in the in situ oral apparatus and somatic basal bodies in situ. Quantitative analysis of gold particle distribution was used to demonstrate the specificity of the antibodies for the basal body-cage complex and to show that non-specific binding of the antibodies was negligible. Preadsorption of the antibody with muscle actin effectively eliminated the capacity of the antibody to bind to proteins on immunoblots and to basal body structures with the immunogold labeling technique. These results provide evidence for actin in the basal body-cage complex and raise the possibility of a contractile system associated with basal bodies.

scholarly journals Fluorescence In Situ Hybridization (FISH) Analysis of the Locations of the Oligonucleotides 5S rDNA, (AGGGTTT)3, and (TTG)6 in Three Genera of Oleaceae and Their Phylogenetic Framework

Genes ◽  
2019 ◽  
Vol 10 (5) ◽  
pp. 375 ◽  
Author(s):  
Xiaomei Luo ◽  
Juncheng Liu
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
5S Rdna ◽  
Fish Analysis ◽  
Cytogenetic Map ◽  
Ligustrum Lucidum ◽  
Starting Point ◽  
Central Regions ◽  
Almost All

We report the cytogenetic map for a collection of species in the Oleaceae, and test similarities among the karyotypes relative to their known species phylogeny. The oligonucleotides 5S ribosomal DNA (rDNA), (AGGGTTT)3, and (TTG)6 were used as fluorescence in situ hybridization (FISH) probes to locate the corresponding chromosomes in three Oleaceae genera: Fraxinus pennsylvanica, Syringa oblata, Ligustrum lucidum, and Ligustrum × vicaryi. Forty-six small chromosomes were identified in four species. (AGGGTTT)3 signals were observed on almost all chromosome ends of four species, but (AGGGTTT)3 played no role in distinguishing the chromosomes but displayed intact chromosomes and could thus be used as a guide for finding chromosome counts. (TTG)6 and 5S rDNA signals discerned several chromosomes located at subterminal or central regions. Based on the similarity of the signal pattern (mainly in number and location and less in intensity) of the four species, the variations in the 5S rDNA and (TTG)6 distribution can be ordered as L. lucidum < L. × vicaryi < F. pennsylvanica < S. oblata. Variations have observed in the three genera. The molecular cytogenetic data presented here might serve as a starting point for further larger-scale elucidation of the structure of the Oleaceae genome, and comparison with the known phylogeny of Oleaceae family.

scholarly journals Explaining the ocean's richest biodiversity hotspot and global patterns of fish diversity

2018 ◽  
Vol 285 (1888) ◽  
pp. 20181314 ◽  
Author(s):  
Elizabeth Christina Miller ◽  
Kenji T. Hayashi ◽  
Dongyuan Song ◽  
John J. Wiens
Keyword(s):  
Large Scale ◽  
Fish Diversity ◽  
Relative Importance ◽  
Diversification Rates ◽  
Marine Habitats ◽  
History Of ◽  
Richness Patterns ◽  
Colonization Time ◽  
Almost All

For most marine organisms, species richness peaks in the Central Indo-Pacific region and declines longitudinally, a striking pattern that remains poorly understood. Here, we used phylogenetic approaches to address the causes of richness patterns among global marine regions, comparing the relative importance of colonization time, number of colonization events, and diversification rates (speciation minus extinction). We estimated regional richness using distributional data for almost all percomorph fishes (17 435 species total, including approximately 72% of all marine fishes and approximately 33% of all freshwater fishes). The high diversity of the Central Indo-Pacific was explained by its colonization by many lineages 5.3–34 million years ago. These relatively old colonizations allowed more time for richness to build up through in situ diversification compared to other warm-marine regions. Surprisingly, diversification rates were decoupled from marine richness patterns, with clades in low-richness cold-marine habitats having the highest rates. Unlike marine richness, freshwater diversity was largely derived from a few ancient colonizations, coupled with high diversification rates. Our results are congruent with the geological history of the marine tropics, and thus may apply to many other organisms. Beyond marine biogeography, we add to the growing number of cases where colonization and time-for-speciation explain large-scale richness patterns instead of diversification rates.

Localization and In Situ Activities of Homoacetogenic Bacteria in the Highly Compartmentalized Hindgut of Soil-Feeding Higher Termites (Cubitermes spp.)

1999 ◽  
Vol 65 (10) ◽  
pp. 4497-4505 ◽  
Author(s):  
Anne Tholen ◽  
Andreas Brune
Keyword(s):  
Detection Limit ◽  
Companion Paper ◽  
Homoacetogenic Bacteria ◽  
Appl Environ ◽  
Partial Pressures ◽  
Almost All ◽  
Stimulation Of

ABSTRACT Methanogenesis and homoacetogenesis occur simultaneously in the hindguts of almost all termites, but the reasons for the apparent predominance of methanogenesis over homoacetogenesis in the hindgut of the humivorous species is not known. We found that in gut homogenates of soil-feeding Cubitermes spp., methanogens outcompete homoacetogens for endogenous reductant. The rates of methanogenesis were always significantly higher than those of reductive acetogenesis, whereas the stimulation of acetogenesis by the addition of exogenous H2 or formate was more pronounced than that of methanogenesis. In a companion paper, we reported that the anterior gut regions of Cubitermes spp. accumulated hydrogen to high partial pressures, whereas H2 was always below the detection limit (<100 Pa) in the posterior hindgut, and that all hindgut compartments turned into efficient H2 sinks when external H2 was provided (D. Schmitt-Wagner and A. Brune, Appl. Environ. Microbiol. 65:4490–4496, 1999). Using a microinjection technique, we found that only the posterior gut sections P3/4a and P4b, which harbored methanogenic activities, formed labeled acetate from H14CO3 −. Enumeration of methanogenic and homoacetogenic populations in the different gut sections confirmed the coexistence of both metabolic groups in the same compartments. However, the in situ rates of acetogenesis were strongly hydrogen limited; in the P4b section, no activity was detected unless external H2 was added. Endogenous rates of reductive acetogenesis in isolated guts were about 10-fold lower than the in vivo rates of methanogenesis, but were almost equal when exogenous H2 was supplied. We conclude that the homoacetogenic populations in the posterior hindgut are supported by either substrates other than H2 or by a cross-epithelial H2transfer from the anterior gut regions, which may create microniches favorable for H2-dependent acetogenesis.

Sign in / Sign up

Export Citation Format

Share Document