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Patterns of cell division and expression of asymmetric cell fate determinants in postembryonic neuroblast lineages of Drosophila.

We have studied the division of postembryonic neuroblasts (Nbs) in the outer proliferation center (OPC) and central brain anlagen of Drosophila. We focused our attention on three aspects of these processes: the pattern of cellular division, the topological orientation of those divisions, and the expression of asymmetric cell fate determinants. Although larval Nbs are of embryonic origin, our results indicate that their properties appear to be modified during development. Several conclusions can be summarized: (i) In early larvae, Nbs divide symmetrically to give rise to two Nbs while in the late larval brain most Nbs divide asymmetrically to bud off an intermediate ganglion mother cell (GMC) that very rapidly divides into two ganglion cells (GC). (ii) Symmetric and asymmetric divisions of OPC Nbs show tangential and radial orientations, respectively. (iii) This change in the pattern of division correlates with the expression of inscuteable, which is apically localized only in asymmetric divisions. (iv) The spindle of asymmetrically dividing Nb is always oriented on an apical-basal axis. (v) Prospero does not colocalize with Miranda in the cortical crescent of mitotic Nbs. (vi) Prospero is transiently expressed in one of the two sibling GCs generated by the division of GMCs. The implications of these results on cell fate specification and differentiation of adult brain neurons are discussed.

Animals↗

Influence of fusicoccin on the control of cell division by auxins.

An indirect stimulation of cell division by fusicoccin is demonstrated. The distribution of 2,4-dichlorophenoxyacetic acid molecules between Acer pseudoplatanus cells and their culture medium is strongly modified by a fusicoccin treatment, through the extracellular acidification induced by the toxin. As a consequence, a stimulation of cell division is observed when the intracellular auxin concentration is sufficiently increased to reach the threshold control level (Leguay JJ, J Guern 1977 Plant Physiol 60: 265-270).Independently of this indirect action on cell division, the number of cells per cluster is decreased and the volume of the cells increased in fusicoccintreated cells which show a typical enlargement response.

Journal Article↗

Heterotrimeric G proteins and regulation of size asymmetry during cell division.

The generation of daughter cells of different fate and size depends on the orientation, positioning and morphology of the mitotic spindle. In both C. elegans and Drosophila, heterotrimeric G proteins have emerged as central and conserved regulators of this process. Although the same molecular players are involved in worms and flies, there are clear differences in the mechanisms used. Interestingly, recent work in mammalian cells suggests that heterotrimeric G proteins may control spindle positioning in higher organisms during symmetric and asymmetric cell divisions.

Animals↗

Cell division and cellular morphology of the chick retinal pigmented epithelial cells in culture: a time-lapse analysis.

OBJECTIVE: To investigate the patterns of cell division, movement and shape during early stages of development of the chick embryo retinal pigmented epithelial (RPE) cells and to evaluate the morphology of dissociated embryonic cells with regard to their proliferation capacity. METHODS: We conducted this study at the Department of Histology and Embryology, Celal Bayar University, Manisa, Turkey, between 2002 and 2003. We isolated the cells from chick embryos. We analyzed the images of the embryonic cells originated from neuroepithelia using a computer-based time-lapse acquisition system attached to a differential interference contrast microscope. RESULTS: Retinal pigmented epithelial cells, despite being dissociated, depict a colony-type growth. Cells in the periphery of the colony and those outside the colony showed a tendency to proliferate and migrate and retained contact with the neighboring cells during division. Characteristics of cytokinesis were separation from the neighboring cell while retaining an attachment point, became rounded, moved up and started to shake and ascend to disseminate to the substrate to complete the division. The round-up stage was non-significantly shorter when the cell was closer to the center of the colony. Cells that were in the periphery of, or outside the colony had a round-up time of over one hour while cytokinesis-to-adhesion time was around 5 minutes. However, when we found the cells in the center of the colony, the times were half-an-hour and 1.5 hours for the daughter cells, a 2-fold difference between daughter cells with regard to the duration of attachment. CONCLUSION: Cell division, migration and proliferation are complex procedures influenced by growth factors, cell adhesion, matrix molecules underneath and the signal mechanisms and can be studied in detail using time-lapse microscopy, immunohistochemistry and confocal microscopy.

Animals↗

Growth and cell division of Mycobacterium avium.

The rates of cell division and of protein, DNA and RNA synthesis upon transition of Mycobacterium avium to and from rich medium were examined. The changes in cell morphology (elongation) were also examined by optical and electron microscopy. Upon transfer from poor to rich medium, the rate of synthesis of RNA increased rapidly, followed by an increase in protein synthesis within 3 h and by an increase in DNA synthesis within 7 h; cell division began after a lag of about 10 h. Upon transfer from rich to poor medium, the preshift rates for protein and DNA synthesis changed to postshift rates after 3 h and 7 h, respectively; RNA synthesis stopped immediately, there was a transient fall in total RNA, and synthesis was resumed at a new rate only after 24 h. After the period of adjustment to new medium, the bacteria entered the postshift growth in which cell size, the increase in cell mass (absorbance at 650 nm) and viable counts, and the rates of synthesis of protein, DNA and RNA were constant. Ultrastructural examination of elongated cells during the adjustment period showed that they had septa at different stages of formation, but no evidence of fragmentation was found. It was concluded that cell division in M. avium was by binary fission, and that the notion of a life-cycle was not supported by present findings.

Bacterial Proteins↗

Planar cell polarity signalling controls cell division orientation during zebrafish gastrulation.

Oriented cell division is an integral part of pattern development in processes ranging from asymmetric segregation of cell-fate determinants to the shaping of tissues. Despite proposals that it has an important function in tissue elongation, the mechanisms regulating division orientation have been little studied outside of the invertebrates Caenorhabditis elegans and Drosophila melanogaster. Here, we have analysed mitotic divisions during zebrafish gastrulation using in vivo confocal imaging and found that cells in dorsal tissues preferentially divide along the animal-vegetal axis of the embryo. Establishment of this animal-vegetal polarity requires the Wnt pathway components Silberblick/Wnt11, Dishevelled and Strabismus. Our findings demonstrate an important role for non-canonical Wnt signalling in oriented cell division during zebrafish gastrulation, and indicate that oriented cell division is a driving force for axis elongation. Furthermore, we propose that non-canonical Wnt signalling has a conserved role in vertebrate axis elongation, orienting both cell intercalation and mitotic division.

Animals↗

discordia mutations specifically misorient asymmetric cell divisions during development of the maize leaf epidermis.

In plant cells, cytokinesis depends on a cytoskeletal structure called a phragmoplast, which directs the formation of a new cell wall between daughter nuclei after mitosis. The orientation of cell division depends on guidance of the phragmoplast during cytokinesis to a cortical site marked throughout prophase by another cytoskeletal structure called a preprophase band. Asymmetrically dividing cells become polarized and form asymmetric preprophase bands prior to mitosis; phragmoplasts are subsequently guided to these asymmetric cortical sites to form daughter cells of different shapes and/or sizes. Here we describe two new recessive mutations, discordia1 (dcd1) and discordia2 (dcd2), which disrupt the spatial regulation of cytokinesis during asymmetric cell divisions. Both mutations disrupt four classes of asymmetric cell divisions during the development of the maize leaf epidermis, without affecting the symmetric divisions through which most epidermal cells arise. The effects of dcd mutations on asymmetric cell division can be mimicked by cytochalasin D treatment, and divisions affected by dcd1 are hypersensitive to the effects of cytochalasin D. Analysis of actin and microtubule organization in these mutants showed no effect of either mutation on cell polarity, or on formation and localization of preprophase bands and spindles. In mutant cells, phragmoplasts in asymmetrically dividing cells are structurally normal and are initiated in the correct location, but often fail to move to the position formerly occupied by the preprophase band. We propose that dcd mutations disrupt an actin-dependent process necessary for the guidance of phragmoplasts during cytokinesis in asymmetrically dividing cells.

Actins↗

A homologue of the bacterial cell division site-determining factor MinD mediates placement of the chloroplast division apparatus.

BACKGROUND: Chloroplast division in plant cells occurs by binary fission, yielding two daughter plastids of equal size. Previously, we reported that two Arabidopsis homologues of FtsZ, a bacterial protein that forms a cytokinetic ring during cell division, are essential for plastid division in plants, and may be involved in the formation of plastid-dividing rings on both the stromal and cytosolic surfaces of the chloroplast envelope membranes. In bacteria, positioning of the FtsZ ring at the center of the cell is mediated in part by the protein MinD. Here, we identified AtMinD1, an Arabidopsis homologue of MinD, and investigated whether positioning of the plastid-division apparatus at the plastid midpoint might involve a mechanism similar to that in bacteria. RESULTS: Sequence analysis and in vitro chloroplast import experiments indicated that AtMinD1 contains a transit peptide that targets it to the chloroplast. Transgenic Arabidopsis plants with reduced AtMinD1 expression exhibited variability in chloroplast size and number and asymmetrically constricted chloroplasts, strongly suggesting that the plastid-division machinery is misplaced. Overexpression of AtMinD1 inhibited chloroplast division. These phenotypes resemble those of bacterial mutants with altered minD expression. CONCLUSIONS: Placement of the plastid-division machinery at the organelle midpoint requires a plastid-targeted form of MinD. The results are consistent with a model whereby assembly of the division apparatus is initiated inside the chloroplast by the plastidic form of FtsZ, and suggest that positioning of the cytosolic components of the apparatus is specified by the position of the plastidic components.

Amino Acid Sequence↗

Muscle gene activation by induction and the nonrequirement for cell division.

We are investigating the mechanism by which animal cells of an amphibian blastula are induced to differentiate as muscle after contact with blastula vegetal cells. After briefly summarizing previous work on this system, we have asked whether this response of animal cells to vegetal induction requires cell division. Animal and vegetal parts of a blastula were placed in contact with each other, and the resulting conjugates cultured in medium containing a sufficient concentration of colchicine or cytochalasin B to inhibit cell division. Muscle differentiation, as indicated by cardiac actin gene transcription, is induced when cell division is inhibited, though at a substantially reduced rate. However, cytoskeletal actin gene transcription, which does not depend on induction, is also much reduced under the same inhibitory conditions. We conclude that, although the cell division inhibitors seem to reduce all gene transcription, they have no preferential effect on the response to induction, and therefore that this process does not require cytoplasmic or nuclear division.

Actins↗

Using carboxyfluorescein diacetate succinimidyl ester to monitor human NK cell division: analysis of the effect of activating and inhibitory class I MHC receptors.

Human NK cells labelled intracellularly with the fluorescent dye 5- and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) were used to assess the effect of ligating class I MHC receptors on NK cell division. The NK cell lines used in these studies expressed a selection of the killer immunoglobulin-like receptors CD158b and CD158a and the CD94/NKG2 family of C-type lectin receptors. The NK cells were cultured in medium containing recombinant (r)IL-2 and receptors were ligated using plastic bound mAb or using soluble murine IgG mAb and FcRII+ gamma-irradiated murine P815 cells. The results obtained show that ligating class I MHC-activating receptors in either culture system stimulates NK cells to divide. Quantitative analysis of cell division reveals that a substantial loss of NK progenitor cells occurs when NK cell-activating receptors are ligated using plastic bound mAb, consistent with concomitant activation-induced cell death. By contrast, progenitor cell loss is prevented when activating receptors are ligated using soluble mAb and P815 cells, suggesting a role for cellular costimulation in cell survival. When inhibitory receptors are coligated with activating receptors using soluble mAb and P815 cells, NK cell division is inhibited. These results demonstrate the potential importance of the activating and inhibitory class I MHC receptors in regulating NK cell proliferation.

Animals↗

Isolation of Chinese hamster ovary cell mutants requiring the continuous presence of taxol for cell division.

Chinese hamster ovary (CHO) cell mutants resistant to the cytotoxic effects of taxol and requiring the drug for normal growth were isolated in a single step. One of these mutant cell lines, Tax-18, fails to divide in the absence of taxol; instead, the cells become larger, rounder, flatter, and multinucleated. Analysis by flow cytometry indicates that during taxol deprivation there is an accumulation of cells in G2 + M phase but that the cells are able to leak through the block in the absence of cell division and further increase their DNA content beyond the tetraploid amount. This interpretation is confirmed by karyotype analysis and by time-lapse studies that show cells rounded for mitosis two to five times longer than in wild-type cultures or in Tax-18 cultures grown in taxol. The cells finally attempt to undergo cytokinesis, fail, and spread out again, but as larger cells than before. Tax-18 has a normal growth rate and morphology when grown in taxol even at concentrations three to five times below the selecting concentration of the drug. The cells, however, have increased sensitivity to microtubule-disrupting drugs such as colcemid, griseofulvin, and D2O. The mutation for taxol auxotrophy behaves recessively in somatic cell hybridization experiments, and the phenotypic reversion rate is approximately 10(-5) in a nonmutagenized population. Both alpha- and beta-tubulin are present in apparently normal amounts and with normal electrophoretic mobilities on two-dimensional gels. The results suggest that Tax-18 lacks a factor necessary for mitosis and that taxol may be able to substitute for this factor.

Alkaloids↗

The ROOT MERISTEMLESS1/CADMIUM SENSITIVE2 gene defines a glutathione-dependent pathway involved in initiation and maintenance of cell division during postembryonic root development.

Activation of cell division in the root apical meristem after germination is essential for postembryonic root development. Arabidopsis plants homozygous for a mutation in the ROOT MERISTEMLESS1 (RML1) gene are unable to establish an active postembryonic meristem in the root apex. This mutation abolishes cell division in the root but not in the shoot. We report the molecular cloning of the RML1 gene, which encodes the first enzyme of glutathione (GSH) biosynthesis, gamma-glutamylcysteine synthetase, and which is allelic to CADMIUM SENSITIVE2. The phenotype of the rml1 mutant, which was also evident in the roots of wild-type Arabidopsis and tobacco treated with an inhibitor of GSH biosynthesis, could be relieved by applying GSH to rml1 seedlings. By using a synchronized tobacco cell suspension culture, we showed that the G(1)-to-S phase transition requires an adequate level of GSH. These observations suggest the existence of a GSH-dependent developmental pathway essential for initiation and maintenance of cell division during postembryonic root development.

Alleles↗

Evidence for polar positional information independent of cell division and nucleoid occlusion.

We present evidence that, in Escherichia coli, polar positional information is present at midcell independent of known cell division factors. In filamented cells, IcsA, which is normally polar, localizes at or near potential cell division sites. Because the cell pole is derived from the septum, the sites to which IcsA localizes in filaments correspond to future poles. IcsA localization to these sites is independent of FtsZ, MinCDE, septation, and nucleoid occlusion, indicating that positional information for the future pole is independent of cell division and chromosome positioning. Upon IcsA localization to these sites, septation is inhibited, suggesting that IcsA recognition of this polar positional information may influence cell division.

Bacterial Proteins↗

Cell division in Escherichia coli: analysis of double mutants for filamentation and abnormal septation of deoxyribonucleic acid-less cells.

The link between chromosome termination, initiation of cell division, and choice of division sites was studied in Escherichia coli by preparing double mutants. Hybrid mutants containing div52-ts, a cell division initiation mutation, and min, mutations which affect the choice of division sites resulting in the septation of minicells, were characterized. The mutants produced minicells and normal cells coordinately under all conditions studied, although the fraction of minicells is half that of the parental minicell strain. The mutant gradually stopped dividing at both the median and minicell septation sites when transferred from 30 to 41 C in rich medium. A synchronous cell division of filaments was induced 15 min after addition of chloramphenicol to the medium, even at 41 C. Divisions were observed at both normal and minicell sites. These results indicate that div52-ts and min functions share a common step in a cell division pathway. A double mutant containing div52-ts and div27-ts, a dnaB mutant which divides in the absence of DNA synthesis, was characterized. The mutant continues to divide after a shift to the high temperature, although at a reduced rate. The behavior of this hybrid mutant suggests a hypothesis that the chromosome termination signal and div52-ts division initiation signal act on a single membrane site which is altered in div27-ts strains.

Cell Count↗

Electron microscopic observations of cell division in Mycobacterium vaccae V1.

Cell division of Mycobacterium vaccae was initiated by deposition of new wall material in the cross wall. The surface layers of the old wall remained continuous until septum formation was complete. Subsequently, rupture of the outer cell wall layers occurred circumferentially, leaving rings on the cell wall. The two daughter cells remained connected with each other at the new pole and bent to form V-shaped structures at the connecting point.

Cell Division↗

Cell divisions in the drosophila embryonic mesoderm are repressed via posttranscriptional regulation of string/cdc25 by HOW.

BACKGROUND: Cell-cycle progression is tightly regulated during embryonic development. In the Drosophila early embryo, the levels of String/Cdc25 define the precise timing and sites of cell divisions. However, cell-cycle progression is arrested in the mesoderm of gastrulating embryos despite a positive transcriptional string/cdc25 activation provided by the mesoderm-specific action of Twist. Whereas String/Cdc25 is negatively regulated by Tribbles in the mesoderm at these embryonic stages, the factor(s) controlling string/cdc25 mRNA levels has yet to be elucidated. RESULTS: Here, we show that the repressor isoform of the Drosophila RNA binding protein Held Out Wing [HOW(L)] is required to inhibit mesodermal cell division during gastrulation. Embryos mutant for how exhibited an excess of cell divisions, leading to delayed mesoderm invagination. The levels of the mitotic activator string/cdc25 mRNA in these embryos were significantly elevated. Protein-RNA precipitation experiments show that HOW(L) binds string/cdc25 mRNA. Overexpression of HOW(L) in Schneider cells reduces specifically the steady-state mRNA levels of a gfp reporter fused to string/cdc25 untranslated region (3'UTR). CONCLUSIONS: Our results suggest that in wild-type embryos, string/cdc25 mRNA levels are downregulated by the repressor isoform HOW(L), which binds directly to string/cdc25 mRNA and regulates its degradation. Thus, we are proposing a novel posttranscriptional mechanism controlling cell-cycle progression in the Drosophila embryo.

Animals↗

Increased cell division but not thymic dysfunction rapidly affects the T-cell receptor excision circle content of the naive T cell population in HIV-1 infection.

Recent thymic emigrants can be identified by T cell receptor excision circles (TRECs) formed during T-cell receptor rearrangement. Decreasing numbers of TRECs have been observed with aging and in human immunodeficiency virus (HIV)-1 infected individuals, suggesting thymic impairment. Here, we show that in healthy individuals, declining thymic output will affect the TREC content only when accompanied by naive T-cell division. The rapid decline in TRECs observed during HIV-1 infection and the increase following HAART are better explained not by thymic impairment, but by changes in peripheral T-cell division rates. Our data indicate that TREC content in healthy individuals is only indirectly related to thymic output, and in HIV-1 infection is mainly affected by immune activation.

Anti-HIV Agents↗

The Escherichia coli cell division protein FtsW is required to recruit its cognate transpeptidase, FtsI (PBP3), to the division site.

The bacterial cell division protein FtsW has been suggested to perform two functions: stabilize the FtsZ cytokinetic ring, and facilitate septal peptidoglycan synthesis by the transpeptidase FtsI (penicillin-binding protein 3). We show here that depleting Escherichia coli cells of FtsW had little effect on the abundance of FtsZ rings but abrogated recruitment of FtsI to potential division sites. Analysis of FtsW localization confirmed and extended these results; septal localization of FtsW required FtsZ, FtsA, FtsQ, and FtsL but not FtsI. Thus, FtsW is a late recruit to the division site and is essential for subsequent recruitment of its cognate transpeptidase FtsI but not for stabilization of FtsZ rings. We suggest that a primary function of FtsW homologues--which are found in almost all bacteria and appear to work in conjunction with dedicated transpeptidases involved in division, elongation, or sporulation--is to recruit their cognate transpeptidases to the correct subcellular location.

Artificial Gene Fusion↗