Cortical actin movements during the first cell cycle of the Caenorhabditis elegans embryo

1996 ◽  
Vol 109 (2) ◽  
pp. 525-533 ◽  
Author(s):  
S. Hird
Keyword(s):  
Cell Cycle ◽  
Caenorhabditis Elegans ◽  
Fluorescence Microscopy ◽  
Actin Microfilaments ◽  
Cortical Actin ◽  
Daughter Cells ◽  
Actin Cortex ◽  
Asymmetrically Distributed ◽  
Cytoplasmic Determinants

The first division of the Caenorhabditis elegans embryo is unequal, generating daughter cells with distinct fates. The differences between the cells are believed to result from the partitioning of cytoplasmic determinants during the first cell cycle. Actin microfilaments play a critical, but poorly defined, role in this event. In this paper, the actin cortex in live embryos is studied during cytoplasmic localisation by fluorescently labelling microfilaments in oocytes and then using in vivo fluorescence microscopy to observe their behaviour. This reveals that there is a concerted movement of cortical actin to the anterior of the embryo at the time cytoplasmic localisation takes place. Furthermore, it is demonstrated that endogenous foci of F-actin are asymmetrically distributed following this event; these structures have previously been seen in fixed cortices. A model for the participation of the actin cytoskeleton in cytoplasmic localisation is presented based on these results.

Bcl-2 protein localizes to the chromosomes of mitotic nuclei and is correlated with the cell cycle in cultured epithelial cell lines

1994 ◽  
Vol 107 (2) ◽  
pp. 363-371
Author(s):  
Q.L. Lu ◽  
A.M. Hanby ◽  
M.A. Nasser Hajibagheri ◽  
S.E. Gschmeissner ◽  
P.J. Lu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Epithelial Cell ◽  
Cell Lines ◽  
Cytoplasmic Localization ◽  
Human Epithelial Cell ◽  
Daughter Cells ◽  
Epithelial Cell Lines ◽  
Cell Immortalization

bcl-2 gene expression confers a survival advantage by preventing cells from entering apoptosis. In contrast to the previously described cytoplasmic localization of Bcl-2 in epithelial cells in vivo, in this study we have demonstrated, in a series of human epithelial cell lines, that Bcl-2 also localizes to mitotic nuclei. Both immunocytochemical and immunoelectron microscopical examinations localize this protein to nuclei and in particular to chromosomes. Nuclear Bcl-2 expression in these cell lines is correlated with the cell cycle. There is relatively strong expression during mitosis, most intense during prophase and metaphase, declining in telophase and then the protein becomes undetectable soon after separation of the two daughter cells. The expression and distribution of Bcl-2 is influenced by treatment with excessive thymidine. These results indicate that Bcl-2 may protect the cells from apoptosis occurring during mitosis and suggest a possible role for the protein in cell immortalization.

scholarly journals Kinetochore protein Spindly controls microtubule polarity in Drosophila axons

2020 ◽  
Author(s):  
Urko del Castillo ◽  
Hans-Arno J. Müller ◽  
Vladimir I. Gelfand
Keyword(s):  
Intracellular Transport ◽  
Critical Role ◽  
Cytoplasmic Dynein ◽  
Neuronal Cultures ◽  
Cortical Actin ◽  
Larval Brain ◽  
Primary Neuronal Cultures ◽  
Actin Cortex ◽  
Microtubule Polarity

AbstractMicrotubule polarity in axons and dendrites defines the direction of intracellular transport in neurons. Axons contain arrays of uniformly polarized microtubules with plus-ends facing the tips of the processes (plus-end-out), while dendrites contain microtubules with minus-end-out orientation. It has been shown that cytoplasmic dynein, targeted to cortical actin, removes minus-end-out microtubules from axons. Here we have identified Spindly, a protein known for recruitment of dynein to kinetochores in mitosis, as a key factor required for dynein-dependent microtubule sorting in axons of Drosophila neurons. Depletion of Spindly affects polarity of axonal microtubules in vivo and in primary neuronal cultures. In addition to these defects, depletion of Spindly in neurons causes major collapse of axonal patterning in the third-instar larval brain as well as severe coordination impairment in adult flies. These defects can be fully rescued by full-length Spindly, but not by variants with mutations in its dynein-binding site. Biochemical analysis demonstrated that Spindly binds F-actin, suggesting that Spindly serves as a link between dynein and cortical actin in axons. Therefore, Spindly plays a critical role during neurodevelopment by mediating dynein-driven sorting of axonal microtubules.Significance StatementNeurons send and receive electrical signals through long microtubule-filled neurites called axons and dendrites. One of the main structural differences between axons and dendrites is how their microtubules are organized. Axons contains microtubules with their plus-ends out while microtubules in dendrites are organized with mixed or plus-end-in orientation. Dynein, the main minus-end microtubule motor, anchored to cortical actin filaments in the axons is responsible for the uniform microtubule polarity in axons. However, it is unknown how dynein is recruited to the actin cortex in axons. The major finding of this work is that Spindly, a protein involved in anchoring dynein to kinetochores during cell division, has a second important function in interphase cells recruiting dynein to the actin cortex in axons.

The shape of mitochondria and the number of mitochondrial nucleoids during the cell cycle of Euglena gracilis

1989 ◽  
Vol 93 (3) ◽  
pp. 565-570
Author(s):  
YASUKO HAYASHI ◽  
KATSUMI UEDA
Keyword(s):  
Cell Cycle ◽  
Fluorescence Microscopy ◽  
Cell Volume ◽  
Ethidium Bromide ◽  
Euglena Gracilis ◽  
Daughter Cell ◽  
Initial Value ◽  
Daughter Cells ◽  
Total Length ◽  
Mitochondrial Nucleoids

The shape of mitochondria and the number of mitochondrial nucleoids in Euglena cells were examined throughout the cell cycle by fluorescence microscopy. Both photoheterotrophic and heterotrophic cells contained a network of mitochondria that did not divide into fragments at any stage of the cell cycle. Mitochondrial nucleoids could be clearly detected in the mitochondria by staining with ethidium bromide and with DAPI. Half of the mitochondrial nucleoids entered each daughter cell during cytokinesis. Nucleoids in the newly produced daughter cells increased in number as the cells increased in size. The number of nucleoids reached double the initial value in cells at the stage just prior to mitosis. The total length of the mitochondrial net was proportional to the cell volume.

scholarly journals The Complex Cell Cycle of the Dinoflagellate Protoctist Crypthecodinium Cohnii as Studied In Vivo and by Cytofluorimetry

1991 ◽  
Vol 100 (3) ◽  
pp. 675-682 ◽  
Author(s):  
YVONNE BHAUD ◽  
JEAN-MARIE SALMON ◽  
MARIE-ODILE SOYER-GOBILLARD
Keyword(s):  
Cell Cycle ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
S Phase ◽  
Vegetative Cells ◽  
Crypthecodinium Cohnii ◽  
Daughter Cells ◽  
The Times

The complete cell cycle of the dinoflagellate Crypthecodinium cohnii Biecheler 1938 was observed in vivo in a synchronized heterogeneous population, after DAPI staining of DNA. In a given population, the relative nuclear DNA content in each class of cell was measured using a new numerical image-analysis method that takes into account the total fluorescence intensity (FI), area (A) and shape factor (SF). The visible degree of synchronization of the population was determined from the number of cells with a nuclear content of 1C DNA at ‘synchronization’, time 0. One method of synchronization (method 1), based on the adhesiveness of the cysts, gave no better than 50% synchronization of the population; method 2, based on swimming cells released from cysts cultured on solid medium, gave 73% of cells with the same nuclear DNA content. A scatter plot of data for FI versus A in the first few hours after time 0 showed that the actual degree of synchronization of the population was lower. The length of the C. cohnii cell cycle determined in vivo by light microscopy was 10, 16 or 24 h for vegetative cells giving two, four or eight daughter cells, respectively. Histograms based on the FI measurements showed that in an initially synchronized population observed for 20 h, the times for the first cell cycle were: G1 phase, 6 h; S phase, 1 h 30 min; G2+M, 1h 30 min, with the release of vegetative cells occurring 1 or 2h after the end of cytokinesis. The times for the second cell cycle were G1+S, 3h; G2+M, 2h. FI and A taken together, suggested that the S phase is clearly restricted, as in higher eukaryotes. A and SF, taken together, showed that the large nuclear areas were always in cysts with two or four daughter cells. FI and SF, taken together, showed that the second S phase always occurred after completion of the first nuclear division. Our data concerning the course of the cell cycle in C. cohnii are compared with those from earlier studies, and the control of the number of daughter cells is discussed; this does not depend on the ploidy of the mother cell.

scholarly journals The Role of Actin in Spindle Orientation Changes during the Saccharomyces cerevisiae Cell Cycle

1999 ◽  
Vol 146 (5) ◽  
pp. 1019-1032 ◽  
Author(s):  
Chandra L. Theesfeld ◽  
Javier E. Irazoqui ◽  
Kerry Bloom ◽  
Daniel J. Lew
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Spindle Orientation ◽  
Cortical Actin ◽  
Yeast Saccharomyces Cerevisiae ◽  
Daughter Cells ◽  
M Phase ◽  
Actin Cables

In the budding yeast Saccharomyces cerevisiae, the mitotic spindle must align along the mother-bud axis to accurately partition the sister chromatids into daughter cells. Previous studies showed that spindle orientation required both astral microtubules and the actin cytoskeleton. We now report that maintenance of correct spindle orientation does not depend on F-actin during G2/M phase of the cell cycle. Depolymerization of F-actin using Latrunculin-A did not perturb spindle orientation after this stage. Even an early step in spindle orientation, the migration of the spindle pole body (SPB), became actin-independent if it was delayed until late in the cell cycle. Early in the cell cycle, both SPB migration and spindle orientation were very sensitive to perturbation of F-actin. Selective disruption of actin cables using a conditional tropomyosin double-mutant also led to de- fects in spindle orientation, even though cortical actin patches were still polarized. This suggests that actin cables are important for either guiding astral microtubules into the bud or anchoring them in the bud. In addition, F-actin was required early in the cell cycle for the development of the actin-independent spindle orientation capability later in the cell cycle. Finally, neither SPB migration nor the switch from actin-dependent to actin-independent spindle behavior required B-type cyclins.

Transient localized accumulation of actin in Caenorhabditis elegans blastomeres with oriented asymmetric divisions

Development ◽  
1994 ◽  
Vol 120 (8) ◽  
pp. 2317-2328 ◽  
Author(s):  
J.A. Waddle ◽  
J.A. Cooper ◽  
R.H. Waterston
Keyword(s):  
Caenorhabditis Elegans ◽  
Actin Filaments ◽  
Molecular Basis ◽  
Capping Protein ◽  
Daughter Cells ◽  
Microtubule Polymerization ◽  
Microtubule Attachment ◽  
Asymmetric Divisions ◽  
Cytoplasmic Determinants ◽  
Anterior Posterior

During Caenorhabditis elegans embryogenesis, specific cells in the P1 lineage rotate their duplicated centrosome pair onto the anterior-posterior axis; this rotation is correlated with and necessary for a differential inheritance of cytoplasmic determinants in the daughter cells. Centrosome pair rotation is sensitive to inhibitors of actin and microtubule polymerization and may require microtubule attachment to a specific cortical site. We show that actin and the barbed-end binding protein, capping protein, transiently accumulate at this cortical site, possibly by assembly onto persistent remnants of previous cell divisions. Based on these observations, we propose a model for the molecular basis of centrosome rotation that is consistent with the dependence of rotation on actin filaments and microtubules.

scholarly journals Symmetry breaking in reconstituted actin cortices

eLife ◽  
2014 ◽  
Vol 3 ◽  
Author(s):  
Enas Abu Shah ◽  
Kinneret Keren
Keyword(s):  
Symmetry Breaking ◽  
Break Symmetry ◽  
Model Systems ◽  
Cortical Actin ◽  
Artificial System ◽  
Actin Cortex ◽  
The Rich ◽  
Oil Emulsions

The actin cortex plays a pivotal role in cell division, in generating and maintaining cell polarity and in motility. In all these contexts, the cortical network has to break symmetry to generate polar cytoskeletal dynamics. Despite extensive research, the mechanisms responsible for regulating cortical dynamics in vivo and inducing symmetry breaking are still unclear. Here we introduce a reconstituted system that self-organizes into dynamic actin cortices at the inner interface of water-in-oil emulsions. This artificial system undergoes spontaneous symmetry breaking, driven by myosin-induced cortical actin flows, which appears remarkably similar to the initial polarization of the embryo in many species. Our in vitro model system recapitulates the rich dynamics of actin cortices in vivo, revealing the basic biophysical and biochemical requirements for cortex formation and symmetry breaking. Moreover, this synthetic system paves the way for further exploration of artificial cells towards the realization of minimal model systems that can move and divide.

scholarly journals Cofilin, But Not Profilin, Is Required for Myosin-I-Induced Actin Polymerization and the Endocytic Uptake in Yeast

2002 ◽  
Vol 13 (11) ◽  
pp. 4074-4087 ◽  
Author(s):  
Fatima-Zahra Idrissi ◽  
Bianka L. Wolf ◽  
M. Isabel Geli
Keyword(s):  
Plasma Membrane ◽  
Fluorescence Microscopy ◽  
Actin Cytoskeleton ◽  
Actin Filaments ◽  
Actin Polymerization ◽  
Cortical Actin ◽  
Myosin I ◽  
Robust Evidence

Mutations in the budding yeast myosins-I (MYO3 andMYO5) cause defects in the actin cytoskeleton and in the endocytic uptake. Robust evidence also indicates that these proteins induce Arp2/3-dependent actin polymerization. Consistently, we have recently demonstrated, using fluorescence microscopy, that Myo5p is able to induce cytosol-dependent actin polymerization on the surface of Sepharose beads. Strikingly, we now observed that, at short incubation times, Myo5p induced the formation of actin foci that resembled the yeast cortical actin patches, a plasma membrane-associated structure that might be involved in the endocytic uptake. Analysis of the machinery required for the formation of the Myo5p-induced actin patches in vitro demonstrated that the Arp2/3 complex was necessary but not sufficient in the assay. In addition, we found that cofilin was directly involved in the process. Strikingly though, the cofilin requirement seemed to be independent of its ability to disassemble actin filaments and profilin, a protein that closely cooperates with cofilin to maintain a rapid actin filament turnover, was not needed in the assay. In agreement with these observations, we found that like the Arp2/3 complex and the myosins-I, cofilin was essential for the endocytic uptake in vivo, whereas profilin was dispensable.

Hematopoietic Stem Cell Tracking In Vivo: A Comparison of Short-Term and Long-Term Repopulating Cells

Blood ◽  
1999 ◽  
Vol 93 (6) ◽  
pp. 1916-1921 ◽  
Author(s):  
Sophie M. Lanzkron ◽  
Michael I. Collector ◽  
Saul J. Sharkis
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Bone Marrow ◽  
Cell Tracking ◽  
Short Term ◽  
Hematopoietic Stem ◽  
Daughter Cells ◽  
Limiting Dilution

We have previously demonstrated that we could separate long-term repopulating stem cells from cells that provided radioprotection (short-term repopulating cells) on the basis of size and suggested that this might be due to the quiescent nature of long-term repopulating cells. To further define the activity of these populations, we used a dye (PKH26), which incorporates into the membrane of cells and is equally distributed to daughter cells when they divide. We developed an assay, which allowed us to retrieve PKH26+ long-term and short-term repopulating cells in the hematopoietic tissues of the recipients posttransplant. We were able to recover the labeled cells and determine their cell cycle activity, as well as their ability to reconstitute secondary lethally irradiated hosts in limiting dilution. The results of our assay suggest that long-term repopulating cells are quiescent in the bone marrow (BM) 48 hours after transplant. We were able to detect only a few labeled cells in the peripheral blood posttransplant and even though cells homed to both the spleen and BM, more long-term repopulating cells homed to the marrow and only these cells, which homed to the marrow, were capable of reconstituting lethally irradiated secondary hosts long-term.

scholarly journals Antitumor Activity of Ethanolic Extract ofDendrobium formosumin T-Cell Lymphoma: AnIn VitroandIn VivoStudy

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Ritika Prasad ◽  
Biplob Koch
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Antitumor Activity ◽  
Fluorescence Microscopy ◽  
Ethanolic Extract ◽  
Dependent Manner ◽  
Isolation And Characterization ◽  
Dalton’S Lymphoma ◽  
Dalton's Lymphoma

Dendrobium, a genus of orchid, was found to possess useful therapeutic activities like anticancer, hypoglycaemic, antimicrobial, immunomodulatory, hepatoprotective, antioxidant, and neuroprotective activities. The study was aimed to evaluate the anticancer property of the ethanolic extract ofDendrobium formosumon Dalton’s lymphoma.In vitrocytotoxicity was determined by MTT assay, apoptosis was determined by fluorescence microscopy, and cell cycle progression was analysed using flow cytometry;in vivoantitumor activity was performed in Dalton’s lymphoma bearing mice. The IC50value of ethanolic extract was obtained at 350 μg/mL in Dalton’s lymphoma cells. Fluorescence microscopy analysis showed significant increase in apoptotic cell death in dose- and time-dependent manner which was further confirmed through the resulting DNA fragmentation. Further, flow cytometry analysis showed that the ethanolic extract arrests the cells in G2/M phase of the cell cycle. Thein vivoanticancer activity study illustrates significant increase in the survival time of Dalton’s lymphoma bearing mice on treatment with ethanolic extract when compared to control. These results substantiate the antitumor properties of ethanolic extract ofDendrobium formosumand suggest an alternative in treatment of cancer. Further studies are required regarding the isolation and characterization of bioactive components along with the analysis of molecular mechanism involved.

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