Mother Cell Nuclei Research Articles

4D Chromatin dynamics in cycling cells

This live cell study of chromatin dynamics in four dimensions (space and time) in cycling human cells provides direct evidence for three hypotheses first proposed by Theodor Boveri in seminal studies of fixed blastomeres from Parascaris equorum embryos: (I) Chromosome territory (CT) arrangements are stably maintained during interphase. (II) Chromosome proximity patterns change profoundly during prometaphase. (III) Similar CT proximity patterns in pairs of daughter nuclei reflect symmetrical chromosomal movements during anaphase and telophase, but differ substantially from the arrangement in mother cell nucleus. Hypothesis I could be confirmed for the majority of interphase cells. A minority, however, showed complex, rotational movements of CT assemblies with large-scale changes of CT proximity patterns, while radial nuclear arrangements were maintained. A new model of chromatin dynamics is proposed. It suggests that long-range DNA-DNA interactions in cell nuclei may depend on a combination of rotational CT movements and locally constrained chromatin movements.

4D Chromatin dynamics in cycling cells: Theodor Boveri's hypotheses revisited

This live cell study of chromatin dynamics in four dimensions (space and time) in cycling human cells provides direct evidence for three hypotheses first proposed by Theodor Boveri in seminal studies of fixed blastomeres from Parascaris equorum embryos: (I) Chromosome territory (CT) arrangements are stably maintained during interphase. (II) Chromosome proximity patterns change profoundly during prometaphase. (III) Similar CT proximity patterns in pairs of daughter nuclei reflect symmetrical chromosomal movements during anaphase and telophase, but differ substantially from the arrangement in mother cell nucleus. Hypothesis I could be confirmed for the majority of interphase cells. A minority, however, showed complex, rotational movements of CT assemblies with large-scale changes of CT proximity patterns, while radial nuclear arrangements were maintained. A new model of chromatin dynamics is proposed. It suggests that long-range DNA-DNA interactions in cell nuclei may depend on a combination of rotational CT movements and locally constrained chromatin movements.

Nonsense-Mediated Decay of ash1 Nonsense Transcripts in Saccharomyces cerevisiae

Nonsense-mediated mRNA decay (NMD) performs two functions in eukaryotes, one in controlling the expression level of a substantial subset of genes and the other in RNA surveillance. In the vast majority of genes, nonsense mutations render the corresponding transcripts prone to surveillance and subject to rapid degradation by NMD. To examine whether some classes of nonsense transcripts escape surveillance, we asked whether NMD acts on mRNAs that undergo subcellular localization prior to translation. In Saccharomyces cerevisiae, wild-type ASH1 mRNA is one of several dozen transcripts that are exported from the mother-cell nucleus during mitotic anaphase, transported to the bud tip on actin cables, anchored at the bud tip, and translated. Although repressed during transport, translation is a prerequisite for NMD. We found that ash1 nonsense mutations affect transport and/or anchoring independently of NMD. The nonsense transcripts respond to NMD in a manner dependent on the position of the mutation. Maximal sensitivity to NMD occurs when transport and translational repression are simultaneously impaired. Overall, our results suggest a model in which ash1 mRNAs are insensitive to NMD while translation is repressed during transport but become sensitive once repression is relieved.

The colocalization transition of homologous chromosomes at meiosis

Meiosis is the specialized cell division required in sexual reproduction. During its early stages, in the mother cell nucleus, homologous chromosomes recognize each other and colocalize in a crucial step that remains one of the most mysterious of meiosis. Starting from recent discoveries on the system molecular components and interactions, we discuss a statistical mechanics model of chromosome early pairing. Binding molecules mediate long-distance interaction of special DNA recognition sequences and, if their concentration exceeds a critical threshold, they induce a spontaneous colocalization transition of chromosomes, otherwise independently diffusing.

Local Activation of Yeast ASH1 mRNA Translation through Phosphorylation of Khd1p by the Casein Kinase Yck1p

In S. cerevisiae, the ASH1 mRNA is localized at the bud tip of late-anaphase cells, resulting in the exclusive sorting of Ash1p to the daughter cell nucleus. While the mechanism behind the localization of this transcript has been well studied, the regulation of its translation is still poorly understood. We now report that the RNA binding protein Khd1 interacts with the ASH1 mRNA localization element E1 and with the C-terminal domain of eIF4G1 to regulate the translation of this transcript. Khd1p reduces translation initiation on the ASH1 mRNA and diminishes Ash1p leakage into the mother cell nucleus. Furthermore, we show that the casein kinase Yck1p phosphorylates Khd1p at the plasma membrane, disrupting the Khd1p-RNA complex and releasing its translational repression on the ASH1 mRNA. This study reveals how, by linking mRNA sorting and translational activation, Khd1p and Yck1p regulate the spatiotemporal expression of a cell fate determinant.

Cytoskeletal asymmetry in Zea mays subsidiary cell mother cells: A monopolar prophase microtubule half‐spindle anchors the nucleus to its polar position

Double labeling of microtubules and actin filaments revealed that in prophase subsidiary mother cells of Zea mays a monopolar prophase microtubule "half-spindle" is formed, which lines the nuclear hemisphere distal to the inducing guard mother cell. The nuclear hemisphere proximal to the guard mother cell is lined by an F-actin cap, consisting of a cortical F-actin patch and actin filaments originating from it. The microtubules of the "half-spindle" decline from the nuclear surface and terminate to the preprophase microtubule band. After disintegration of the latter, a bipolar metaphase spindle is organized. The polar F-actin cap persists during mitosis and early cytokinesis, extending to the chromosomes and the subsidiary cell daughter nucleus. In oryzalin treated subsidiary mother cells the prophase nuclei move away from the polar site. Cytochalasin B and latrunculin-B block the polar migration of subsidiary mother cell nuclei, but do not affect those already settled to the polar position. The prophase nuclei of latrunculin-B treated subsidiary mother cells are globally surrounded by microtubules, while the division plane of latrunculin-B treated subsidiary mother cells is misaligned. The prophase nuclei of brick 1 mutant Zea mays subsidiary mother cells without F-actin patch are also globally surrounded by microtubules. The presented data show that the prophase microtubule "half-spindle"-preprophase band complex anchors the subsidiary mother cell nucleus to the polar cell site, while the polar F-actin cap stabilizes the one metaphase spindle pole proximal to the inducing guard mother cell.

Chromosome order in HeLa cells changes during mitosis and early G1, but is stably maintained during subsequent interphase stages.

Whether chromosomes maintain their nuclear positions during interphase and from one cell cycle to the next has been controversially discussed. To address this question, we performed long-term live-cell studies using a HeLa cell line with GFP-tagged chromatin. Positional changes of the intensity gravity centers of fluorescently labeled chromosome territories (CTs) on the order of several μm were observed in early G1, suggesting a role of CT mobility in establishing interphase nuclear architecture. Thereafter, the positions were highly constrained within a range of ∼1 μm until the end of G2. To analyze possible changes of chromosome arrangements from one cell cycle to the next, nuclei were photobleached in G2 maintaining a contiguous zone of unbleached chromatin at one nuclear pole. This zone was stably preserved until the onset of prophase, whereas the contiguity of unbleached chromosome segments was lost to a variable extent, when the metaphase plate was formed. Accordingly, chromatin patterns observed in daughter nuclei differed significantly from the mother cell nucleus. We conclude that CT arrangements were stably maintained from mid G1 to late G2/early prophase, whereas major changes of CT neighborhoods occurred from one cell cycle to the next. The variability of CT neighborhoods during clonal growth was further confirmed by chromosome painting experiments.

Nuclear DNA amount during sporogenesis and gametogenesis in sexual and aposporous buffelgrass

Nuclear DNA amounts (C values) were measured in Feulgen-stained sections of anthers and ovules of sexual plant B-2s (genotype aaaa) and aposporous cultivar Higgins (genotype AAaa) of buffelgrass (Pennisetum ciliare). The mass of the unreplicated nuclear genome of a gamete equals 1C DNA. In both lines, pollen mother cell nuclei were 4C before leptotene; anther wall, dyad, 1-nucleate pollen, and generative cell nuclei were 2C; microspore tetrad, enlarging microspore, and sperm nuclei were 1C. The tapetum persisted as uninucleate cells with 4C DNA. Archespores (2-4C) of both lines initiated meiosis to form megaspore tetrad nuclei with 1-2C DNA. In B-2s, chalazal megaspores (2-4C) formed reduced 8-nucleate Polygonum type embryo sacs, and sacs at 2- and 4-nucleate stages showed distributions with peaks near C1 and C2, corresponding to G1 and G2 cell cycle phases; this is characteristic of active mitosis. Nuclei of 8-nucleate sacs and of eggs and polars were 1C, indicating chromosomes were not duplicated before fertilization. Antipodal nuclei had levels from 1 to 36C, possibly due to polyteny or endopolyploidy. In Higgins, aposporous initials and 2-nucleate embryo sacs showed bimodal distributions of 2n nuclei with peaks at 2C and 4C DNA. Nuclei of newly formed 4-nucleate Panicum type aposporous sacs and of polars were 2C; aposporous eggs stained too faintly for reliable measurement.

Indiscriminate synapsis in achiasmate Allium fistulosum L. (Liliaceae)

ABSTRACT Seedlings of Allium fistulosum (2n=2x=16) were treated with aqueous colchicine with the intention of inducing tetraploidy. One treated, but undoubled, diploid mutant is described which consistently fails to form any chiasmata at diakinesis and metaphase I of meiosis. Electron microscopy of whole-mount surface-spread synaptonemal complex complements of pollen mother cell nuclei revealed that the achiasmate condition is probably due not only to the failure to complete synapsis, but also to the indiscriminate way in which the chromosomes form synaptonemal complexes during meiotic prophase. Synapsis begins and progresses with complete disregard to homology, with frequent exchanges of pairing partners resulting in the formation of multiple associations comprising heterologous chromosomes. Intrachromosomal synapsis is also evident as fold-back loops. Up to 78% of lateral element length is incorporated into synaptonemal complex, the morphology of which is not unlike that of normal A. fistulosum and other Allium species described previously. However, all the synaptonemal complexes are ineffective in terms of supporting chiasmata, since 16 univalents enter metaphase I and disjoin irregularly at anaphase I. The mutant is as a consequence completely male sterile. The synaptic behaviour observed confirms that the recognition of homology is an independent process and not a prerequisite for synaptonemal complex formation. It is hoped this mutant will be a valuable tool for probing the molecular basis of homology.

Elimination of synaptonemal complex irregularities in a Lolium hybrid

The diploid hybrid ryegrass, Lolium temulentum × L. perenne (2n = 14), contains two sets of chromosomes which are structurally and genetically dissimilar. Whole-mount surface spreading of synaptonemal complexes from 35 pollen mother cell nuclei shows that chromosome pairing during meiotic prophase is highly irregular, with homoeologues and heterologues pairing together to form multivalents. These are eliminated, however, during meiotic prophase in favour of homoeologous bivalents. In addition, the length differential between homoeologous chromosomes is accommodated in a coordinated fashion and is absorbed into the structure of the synaptonemal complexes. The hybrid thus displays a remarkable capacity to eliminate synaptonemal complex irregularities and to produce homoeologous bivalents, which are functionally and morphologically almost indistinguishable from their homologous counterparts in the parents.

Chromosome pairing in a Lolium temulentum X Lolium perenne diploid hybrid with a low chiasma frequency

Despite an average difference of about 50% in DNA amount, homoeologous chromosomes pair effectively at first metaphase in the diploid interspecific hybrid between Lolium temulentum and Lolium perenne. However, in the presence of accessory B chromosomes and "diploidising genes" pairing at metaphase I is severely reduced. Reconstruction of serial electron micrographs through pollen mother cell nuclei show that synaptonemal complexes are formed at pachytene between not only homoeologous but also non-homologous chromosome segments resulting in multivalent formation. These associations are largely ineffective in terms of chiasma formation and degenerate at late pachytene. It is highly probable that the pairing determinants exercise their control on chromosome pairing largely by prohibiting the siting of crossovers in homoeologously paired chromosome segments.

Synaptonemal complex and recombination nodules in rye (Secale cereale)

Meiotic prophase in rye was investigated by serial-section reconstruction of pollen mother cell nuclei. In the mid-late zygotene nucleus, all lateral elements were continuous from telomere to telomere, and 9–20 pairing initiation sites per bivalent were observed. Chromosome and bivalent interlockings detected during zygotene were resolved at early pachytene when pairing was completed. In the three pachytene nuclei, the relative synaptonemal complex (SC) lengths and arm ratios were found to be in good correlation with light microscopic data of pachytene bivalents. Spatial tracing of the bivalents showed that they occupy separate areas in the nucleus. Three types of recombination nodules were observed: large, ellipsoidal and small nodules at early pachytene and irregularly shaped nodules mainly associated with chromatin at late pachytene. Their number and position along the bivalents correlated well with the number and distribution of chiasmata. The classification of the seven bivalents was based on arm ratio and heterochromatic knob distribution.

Structure and development of normal and abnormal stomata in the seedlings of some Cruciferae

The structure and ontogeny of normal and abnormal stomata in the seedlings of 16 species of the Cruciferae have been presented. The mature stomata are anomocytic, paracytic, anisocytic, helicocytic and with a single subsidiary cell. The ontogeny of the anisocytic, paracytic, helicocytic and stoma with a single subsidiary cell conforms to the syndetocheilic or mesogenous, while that of anomocytic to haplocheilic or perigenous type. Several types of aberrant stomatal formations such as: single guard cell; contiguoas single-guard cells; contiguous stomatal; degeneration of guard cell/s; persistert stomatal cell; amitotic division of guard mother cell nucleus followed by pore formation; stoma with double pores; binucleate guard cell; division of guard cell; incomplete stoma with unequal guard cells; cytoplasmic connection; ring-shaped division of guard mother cell and uncommon wall thickening have been noticed. Aberrant stomatal developments are naturally occurring and not induced.

Macrosporogenesis and Embryo Development in Ulmus fulva

1. The ovule of Ulmus fulva is anatropous, consisting of two integuments and a massive nucellus which becomes beaklike at maturity. 2. The archesporial cell functions as a macrospore mother cell. 3. The macrospore mother cell nucleus undergoes meiosis, forming four macrospore nuclei in a linear row. 4. No cell plates or walls are formed between the macrospore nuclei. 5. A typical eight-nucleate macrogametophyte is formed as a result of one further nuclear division. 6. The zygote, by transverse divisions, forms a row of three cells. The apical cell of this row develops into the embryo. The two basal cells form the suspensor.

Societies and Academies

Societies and Academies

Meiotic Cytokinesis of Cannabis

1. Counts made in the pollen mother cells of Cannabis sativa show that the reduced number of chromosomes in this species is ten. 2. The reduction division commonly occurs during the period from sunrise to noon. Although mitotic figures can be found in material collected during the afternoon or evening, they are comparatively rare. 3. The heterotypic division is easily studied, and chromosome counts are most easily made during the anaphase of this division. The behavior of the chromosomes is normal, and no unusual features were noted. 4. Little can be learned concerning the individual chromosomes during the homotypic division, because of the massing of cell elements and the rapidity of the division. 5. There is no morphological evidence that any one of the ten chromosomes is a sex chromosome. 6. The method of cytokinesis is by furrowing. The furrows appear at the periphery of the cytoplasm and at points equidistant from the mother cell nuclei. These furrows extend rapidly to the center of the mother cell, ...