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On the constancy of cell division rate in the root meristem.

This review examines under what circumstances the rate of cell division among cells of the root meristem is known to vary. First, methods are compared that have been used to quantify cell division rate. These can be grouped as being either cytological, in which the rate of accumulation of cells in a particular phase of the cell cycle is determined based on some form of cytological labeling, or kinematic, in which the rate of cell accumulation is determined from the net movement of cells. Then, evidence is reviewed as to whether cell division rates vary between different tissues or cell types, between different positions in the root, or finally between different environments. The evidence is consistent with cells dividing at a constant rate, and well documented examples where cell division rate changes substantially are rare. The constancy of cell division rate contrasts with the number of dividing cells, which varies extensively, and implies that a major point for cell cycle control is governing the exit from the proliferative state at the basal boundary of the meristem.

Biomechanical Phenomena↗

Correlation of cytokinin levels in the endosperms and roots with cell number and cell division activity during endosperm development in rice.

Cell number and cell division activity in rice (Oryza sativa) endosperms are possibly regulated by cytokinin levels in the endosperm and its source in the roots. This study tried to find the possible correlations among them. Six rice genotypes were grown in nutrient solution. Two patterns of endosperm cell division, synchronous and asynchronous, were observed among the genotypes based on the cell division rate of superior and inferior spikelets. Contents of zeatin (Z) + zeatin riboside (ZR) were much higher than those of N6-isopentenyladenine (iP) and N6-isopentenyladenosine (iPR) in both endosperms and roots. Changes in Z + ZR levels in endosperms were significantly correlated with those in roots, and both were very significantly correlated with the cell division rate. Changes in iP + iPR contents in the roots were not significantly correlated with those in the endosperms and the cell division rate. When roots were treated with kinetin, endosperm cell number and grain weight were increased. Such enhancement was more significantly achieved by the root kinetin treatment than by spraying kinetin on leaves and panicles. The results suggest that the cell number and cell division activity in rice endosperms are regulated by cytokinin levels in the endosperm and that root-derived Z + ZR play a pivotal role.

Adenine↗

Wnt signaling and a Hox protein cooperatively regulate psa-3/Meis to determine daughter cell fate after asymmetric cell division in C. elegans.

Asymmetric cell division is a mechanism for achieving cellular diversity. In C. elegans, many asymmetric cell divisions are controlled by the Wnt-MAPK pathway through POP-1/TCF. It is poorly understood, however, how POP-1 determines the specific fates of daughter cells. We found that nob-1/Hox, ceh-20/Pbx, and a Meis-related gene, psa-3, are required for asymmetric division of the T hypodermal cell. psa-3 expression was asymmetric between the T cell daughters, and it was regulated by POP-1 through a POP-1 binding site in the psa-3 gene. psa-3 expression was also regulated by NOB-1 and CEH-20 through a NOB-1 binding sequence in a psa-3 intron. PSA-3 can bind CEH-20 and function after the T cell division to promote the proper fate of the daughter cell. These results indicate that cooperation between Wnt signaling and a Hox protein functions to determine the specific fate of a daughter cell.

Active Transport, Cell Nucleus↗

Evidence that interleukin 2 actively promotes both cell division and immunoglobulin secretion in human B cells.

Three human Epstein-Barr virus transformed B cell lines were seeded at densities below their capacity for autostimulatory growth and exposed to recombinant human interleukin 2 (IL-2). IL-2 was found to stimulate DNA synthesis in all three lines, and the level of immunoglobulin secreted was also greatly increased. The experiments were repeated, but in this case the final cell numbers were established by the MTT method and the immunoglobulin secreted per 1,000 cells calculated. Both of these parameters were significantly elevated, indicating that the higher immunoglobulin levels recorded were due to the rate of secretion having been actively increased by IL-2 and were not merely artifactual, resulting from increased cell growth. Thus, IL-2 can actively promote DNA synthesis and cell division and can also increase immunoglobulin secretion in human B cells.

Antibody Formation↗

ftsW is an essential cell-division gene in Escherichia coli.

In the absence of exogenous promoters, plasmid-mediated complementation of the temperature-sensitive ftsW201 allele requires the presence of the full coding sequence of ftsW plus upstream DNA encompassing the C-terminus of mraY and the full coding sequence of murD. We used molecular and genetic techniques to introduce an insertional inactivation into the chromosomal copy of ftsW, in the presence of the plasmid-borne wild-type ftsW gene under the control of P(BAD). In the absence of arabinose, the ftsW-null strain is not viable, and a shift from arabinose- to glucose-containing liquid medium resulted in a block in division, followed by cell lysis. Immunofluorescence microscopy revealed that in ftsW-null filaments, the FtsZ ring is absent in 50-60% of filaments, whilst between one and three Z-rings per filament can be detected in the remainder of the population, with the majority of these containing only one Z-ring per filament. We also demonstrated that the expression of only ftsWS (the smaller of two ftsW open reading frames) from P(BAD) is sufficient for complementation of the ftsW-null allele. We conclude that FtsW is an essential cell-division protein in Escherichia coli, and that it plays a role in the stabilization of the FtsZ ring during cell division.

Alleles↗

Borrelia burgdorferi ftsZ plays a role in cell division.

ftsZ is essential for cell division in many microorganisms. In Escherichia coli and Bacillus subtilis, FtsZ plays a role in ring formation at the leading edge of the cell division septum. An ftsZ homologue is present in the Borrelia burgdorferi genome (ftsZ(Bbu)). Its gene product (FtsZ(Bbu)) is strongly homologous to other bacterial FtsZ proteins, but its function has not been established. Because loss-of-function mutants of ftsZ(Bbu) might be lethal, the tetR/tetO system was adapted for regulated control of this gene in B. burgdorferi. Sixty-two nucleotides of an ftsZ(Bbu) antisense DNA sequence under the control of a tetracycline-responsive modified hybrid borrelial promoter were cloned into pKFSS1. This construct was electroporated into a B. burgdorferi host strain carrying a chromosomally located tetR under the control of the B. burgdorferi flaB promoter. After induction by anhydrotetracycline, expression of antisense ftsZ RNA resulted in generation of filamentous B. burgdorferi that were unable to divide and grew more slowly than uninduced cells. To determine whether FtsZ(Bbu) could interfere with the function of E. coli FtsZ, ftsZ(Bbu) was amplified from chromosomal DNA and placed under the control of the tetracycline-regulated hybrid promoter. After introduction of the construct into E. coli and induction with anhydrotetracycline, overexpression of ftsZ(Bbu) generated a filamentous phenotype. This suggested interference of ftsZ(Bbu) with E. coli FtsZ function and confirmed the role of ftsZ(Bbu) in cell division. This is the first report of the generation of a B. burgdorferi conditional lethal mutant equivalent by tetracycline-controlled expression of antisense RNA.

Amino Acid Sequence↗

Fetal blood mononuclear cell division in normal and pathological pregnancies.

Fetal blood mononuclear cell division was measured using flow cytometry in 53 normal pregnancies and 51 pathological pregnancies complicated either by anaemia due to red blood cell isoimmunisation (RCI: n = 21), intrauterine growth retardation (SGA: n = 13) or abnormal karyotype (n = 17). In normal pregnancy, mononuclear cell division rates decreased with gestational age from a mean of 1.8% at 18 weeks to 1% at 40 weeks. Furthermore, there was a significant association between cell division and erythroblast count. The rates of cell division and erythroblast count were significantly increased in the chromosomally abnormal fetuses, and significantly decreased in the transfused RCI fetuses compared to the controls. Although the erythroblast count was elevated in the SGA fetuses, the mononuclear cell division was not significantly different from the controls. Fetal blood mononuclear cell division is elevated in early pregnancy and in chromosomally abnormal fetuses, probably as a consequence of increased numbers of circulating haemopoietic precursors. Mononuclear cell division is decreased in transfused RCI fetuses as a consequence of suppressed erythropoiesis. In SGA fetuses, despite the increased erythropoietic stimulation and erythroblastosis, cell division is not increased.

Anemia↗

Differentiation of human embryonic stem cells occurs through symmetric cell division.

Embryonic (ES) stem cells can be expanded indefinitely, yet retain the ability to form all cell types of the body. Here we report that human ES cells differentiate exclusively by symmetric cell division in each of four distinct differentiation conditions examined. This suggests that, in some respects, ES cells more closely resemble precursor or transit amplifying cells rather than adult stem cells.

Cell Differentiation↗

Reduction in the level of intracellular myo-inositol in cultured soybean (Glycine max) cells inhibits cell division.

Although myo-inositol is included in media for the successful growth of plant tissues, the actual requirement of most tissues, including soybean (Glycine max) callus in suspension culture, for myo-inositol has not been demonstrated. We have made use of deoxyglucose to reduce intracellular levels of myo-inositol. Deoxyglucose is phosphorylated to deoxyglucose 6-phosphate, which inhibits L-myo-inositol 1-phosphate synthase, an important enzyme in the synthesis of myo-inositol. Addition of deoxyglucose to the medium resulted in a decrease in the intracellular level of myo-inositol that corresponded with a decrease in cell division. Cell viability was not affected. When myo-inositol was added to cells along with deoxyglucose, cell division was restored, as were intracellular levels of myo-inositol. Addition of myo-inositol had no affect on the uptake or metabolism of deoxyglucose. From these results we propose that myo-inositol has a role in maintaining cell division in soybean callus tissue in suspension culture.

Cell Division↗

Real-time lineage analysis reveals oriented cell divisions associated with morphogenesis at the shoot apex of Arabidopsis thaliana.

Precise knowledge of spatial and temporal patterns of cell division, including number and orientation of divisions, and knowledge of cell expansion, is central to understanding morphogenesis. Our current knowledge of cell division patterns during plant and animal morphogenesis is largely deduced from analysis of clonal shapes and sizes. But such an analysis can reveal only the number, not the orientation or exact rate, of cell divisions. In this study, we have analyzed growth in real time by monitoring individual cell divisions in the shoot apical meristems (SAMs) of Arabidopsis thaliana. The live imaging technique has led to the development of a spatial and temporal map of cell division patterns. We have integrated cell behavior over time to visualize growth. Our analysis reveals temporal variation in mitotic activity and the cell division is coordinated across clonally distinct layers of cells. Temporal variation in mitotic activity is not correlated to the estimated plastochron length and diurnal rhythms. Cell division rates vary across the SAM surface. Cells in the peripheral zone (PZ) divide at a faster rate than in the central zone (CZ). Cell division rates in the CZ are relatively heterogeneous when compared with PZ cells. We have analyzed the cell behavior associated with flower primordium development starting from a stage at which the future flower comprises four cells in the L1 epidermal layer. Primordium development is a sequential process linked to distinct cellular behavior. Oriented cell divisions, in primordial progenitors and in cells located proximal to them, are associated with initial primordial outgrowth. The oriented cell divisions are followed by a rapid burst of cell expansion and cell division, which transforms a flower primordium into a three-dimensional flower bud. Distinct lack of cell expansion is seen in a narrow band of cells, which forms the boundary region between developing flower bud and the SAM. We discuss these results in the context of SAM morphogenesis.

Arabidopsis↗

A set of ftsZ mutants blocked at different stages of cell division in Caulobacter.

FtsZ is required throughout the cell division process in eubacteria and in archaea. We report the isolation of novel mutants of the FtsZ gene in Caulobacter crescentus. Clusters of charged amino acids were changed to alanine to minimize mutations that affect protein folding. Molecular modelling indicated that all the clustered-charged-to-alanine mutations had altered amino acids at the surface of the protein. Of 13 such mutants, four were recessive-lethal, three were dominant-lethal, and six had no discernible phenotype. An FtsZ depletion strain of Caulobacter was constructed to analyse the phenotype of the recessive-lethal mutations and used to show that they blocked cell division at distinct stages. One mutation blocked the initiation of cell division, two mutations blocked cell division randomly, and one mutation blocked both early and late stages of cell division. The effect of the recessive mutations on the subcellular localization of FtsZ was determined. Models to explain the various mutant phenotypes are discussed. This is the first set of recessive alleles of ftsZ blocked at different stages of cell division.

Amino Acid Sequence↗

The fenestrin antigen in submembrane skeleton of the ciliate Tetrahymena thermophila is proposed as a marker of cell polarity during cell division and in oral replacement.

Tetrahymena thermophila cells have two types of polarized morphogenesis: divisional morphogenesis and oral reorganization (OR). The aim of this research is the analysis of cortical patterns of immunostaining during cell division and in OR using previously characterized antibodies against fenestrin and epiplasm B proteins. During cell division, the anarchic field of basal body proliferation of the new developing oral apparatus (AF) showed concomitant strong binding of the fenestrin antigen and withdrawal of a signal of the epiplasm B antigen. At a specific stage, the fenestrin antigen also appeared as a character of the anterior cortex pole, with a co-localized decrease in the detected epiplasm B antigen. The fenestrin antigen also showed a polarity of duplicating basal bodies in ciliary rows. Indirect immunofluorescence and immunogold labeling experiments were performed in the absence and presence of an inhibitor of activity of serine/threonine kinases, 6-dimethylaminopurine (6-DMAP) as an inducer of the oral replacement process. In the presence of 6-DMAP, one class of cells started OR, and some others were trapped and affected in cell division. Both types of cells showed an instability of oral structures and formed enlarged primordial oral fields. These anarchic fields (AFs) bind the fenestrin antigen, with disappearance of epiplasmic antigen staining. Only one protein (about 64 kDa) is detected in western blots by the anti-fenestrin antibody and it accumulated in 6-DMAP-treated cells that are involved in uncompleted morphogenetic activity. At a defined stage of oral development, both during cell division and in OR, the fenestrin antigen served as a marker of polarity of the cell of the anterior pole character.

Adenine↗

Cell division is required for de novo methylation of CpG islands in bladder cancer cells.

Cell division is essential for tumor development and progression. Methylation-mediated silencing caused by aberrant de novo methylation of CpG islands located in the promoter regions of growth regulatory genes occurs frequently in human cancers. We investigated the relationship between cell division and de novo methylation to determine whether de novo methylation can occur in the absence of cell division in cancer cells. We treated T24 bladder carcinoma cells with 5-Aza-2'-deoxycytidine to induce a transient demethylation and then compared the timing and kinetics of remethylation of the p16 gene locus under conditions of either G(0)-G(1) growth arrest induced by serum starvation and confluence or continuous cell proliferation in complete medium. Variable levels of remethylation were detected in CpG poor regions of DNA, as well as repetitive DNA elements in the absence of cell division, yet no remethylation occurred at CpG islands under these conditions. This correlated with continuous expression of p16 protein in these cells. DNA methyltransferase (DNMT)1 and DNMT3b3 proteins were undetectable in 5-Aza-2'-deoxycytidine-treated and untreated nondividing cells, and their mRNA transcripts were down-regulated in these cells. Although DNMT3a mRNA levels were also reduced, they recovered to original levels in nondividing cells after drug treatment. Our results suggest that cell division is required for de novo methylation of CpG islands and that DNMT3a may play a role in methylating CpG poor regions or repetitive DNA elements outside of the S phase of the cell cycle.

Antimetabolites, Antineoplastic↗

Virus-induced transformation without cell division.

Interference with the cell cycle by vinblastine sulfate immediately after cells were infected with Rous sarcoma virus had little effect on the a development of two metabolic changes that occur in transformed cells. These results, along with an earlier demonstration of morphological changes developing in infected nondividing cells, demonstrate that the phenotypic development of the malignant state can occur without the intervention of cell divisions after infection by Rous sarcoma virus.

Animals↗

P-GAP-43 is enriched in horizontal cell divisions throughout rat cortical development.

Asymmetric cell divisions are correlated to neurogenesis in the mammalian cortex and occur often with a horizontal orientation of cell division. However, the molecular mechanisms of spindle orientation or asymmetric cell divisions are not well understood in the developing mammalian central nervous system. Here we show a new molecular marker for horizontally dividing precursors in the mammalian telencephalon. The antibody 2G12 directed against phosphorylated serine of growth associated protein 43 (GAP-43) labels postmitotic neurons and a subset of cells in mitoses in the developing rat telencephalon. 2G12 immunoreactivity was found at a high frequency in mitotic cells dividing parallel to the ventricular surface throughout neurogenesis (embryonic day 13-17) in the cerebral cortex and ganglionic eminence. Interestingly, we detected the same predominance of 2G12 immunoreactivity in horizontally dividing cells in the subventricular zone, the second proliferative layer that has recently been involved in the generation of neurons. Moreover, 2G12 immunostaining is no longer detectable in mitotic cells of the ventricular zones at E21, the onset of gliogenesis in rat telencephalon. These data imply GAP-43 phosphorylation in the phase of neuronal commitment during M-phase and present to our knowledge the first molecular correlate to horizontally dividing precursors in mammalian neurogenesis.

Aging↗

DNA synthesis and cell division related to adipose differentiation of 3T3 cells.

Adipose differentiation of 3T3-F442A cells in surface cultures depends on adipogenic factor present in the culture medium. We found that after stimulation with adipogenic serum, 3T3-F442A cell underwent a burst of DNA synthesis before adipose conversion was manifested by an augmented lipogenic enzyme activity. In differentiating cells, DNA synthesis, judged by a 100-fold higher rate of [3H]thymidine incorporation into TCA-insoluble material, was followed by a 100-fold increase in the activity of glycerophosphate acyltransferase. Cytosine arabinoside, added to the cultures at a concentration of 3 micrograms/ml, exerted 95% inhibition of [3H]thymidine incorporation and also inhibited adipose conversion. The burst of DNA synthesis correlated with a 2.5-fold increase in the amount of DNA and in the number of cells in the culture. The DNA content was the same in differentiated and nondifferentiated cells. We conclude that after the interaction with the adipogenic factor, the cells go through DNA synthesis and cell division essential for adipose conversion.

Adipose Tissue↗

Electron microscopic study of cell surface rings during cell division and morphogenesis of Arthrobacter crystallopoietes.

The whole cell ultrastructure during cell division and morphogenesis of Arthrobacter crystallopoietes was monitored using electron microscopic techniques. Glucose-grown spherical cells were inoculated into succinate-based medium. In this medium, the organism undergoes a morphogenetic cycle consisting of elongation of spheres to rods, exponential growth as rods, and fragmentation of rods to spherical cells. Raised bands or rings that encircled the cells were evident on the cell surface of both sphere- and rod-shaped cells. Many rod-shaped cells possessed two or more rings arranged adjacent to each other in a parallel orientation. At each cell division a new ring was formed on both siblings. However, as predicted by the proposed model of unidirectional cell growth and by maintaining a ring from the previous generation, unequal numbers of rings were observed on sibling cells. Only one ring was visible on most of the spherical inoculum cells, but in some cases a second ring perpendicular to the other ring was observed. Parallel rings were found on spherical cells resulting from fragmentation or reductive cell division of rods during the stationary growth phase. Thus, these spheres could be distinguished from inoculum spheres containing a single ring or perpendicular orientation of rings. The number of rings per cell and arrangement of rings on the cell surface of sibling cells after cell division, but before cell separation, are discussed with respect to cell age, cell division, and sphere-rod-sphere morphogenesis of A. crystallopoietes.

Arthrobacter↗