A mathematical model for cell sorting,migration and shape in the slug stage ofDictyostelium discoideum

A mathematical model for cell sorting and migration in the slug stage of cellular slime moldsDictyostelium discoideum is proposed. Assuming that a slug is a “mixed fluid” of prespore and prestalk cells, a set of equations which describe the dynamics of cell distribution, internal pressure and velocity of hte slug are derived from the balance formula of individual cell movement. These equations are analyzed to obtain the spatial patterns of the two types of cells at dynamical equilibrium and the relationship between the migration velocity and the slug size. The body shape of the elongated slug at the migrating stage is also investigated, taking account of the law of surface tension. The stable shapes of slugs with different volumes are explicity obtaained and the existence of critical size of a slug is suggested.     

Dynamics of Cell Redifferentiation in Dictyostelium mucoroides

During migration of a D. mucoroides slug, prespore cells at the immediate edge of the prespore-prestalk junction advance into the prestalk region in approximately 1.5 min. Redifferentiation of the prespore into prestalk cells occurs during this period. Evidence that the cells redifferentiated required autoradiographic, electron microscopic, and staining techniques. These methods determined that glycoprotein synthesis declined; prespore vesicles, the organelles found specifically in prespore cells, were degraded; and staining was reduced during this developmental transition into prestalk cells.     

Cell Patterning During Slug Migration and Early Culmination in Dictyostelium discoideum

Abstract. The effects of migration and culmination on patterning of presumptive (prespore and prestalk) cells and mature (spore and stalk) cells of D. discoideum were investigated. The ratio of prespore to total cells, as determined by staining with fluorescein-conjugated antispore globulin, was constant (77%) up until 8 h of slug migration, but then decreased to a level (64%) which thereafter remained unchanged during migration. Cells which lost prespore antigen during migration were located in the posterior (prespore) part next to the agar surface.
Upon induction of culmination, however, the ratio of prespore cells quickly increased to the normal level (77%) within 1–2 h. During the transition between migration and culmination prestalk and prespore cells were considerably intermixed within the cell mass, before the normal prestalk-prespore pattern was reestablished at the preculmination (Mexican hat) stage. Spore: stalk ratios within fruiting bodies were normal irrespective of the lengths of slug migration.     

Phagocytosis of algal chloroplasts by digestive gland cells in the photosynthesis-capable slug Elysia timida (Mollusca, Opisthobranchia, Sacoglossa)

Parapodia of the sacoglossan slug Elysia timida were preserved by high-pressure cryofixation during feeding experiments and investigated with transmission electron microscopy. This slug has been known for its long-term retention of active chloroplasts and photosynthesis. We observed different stages of phagocytosis of chloroplast components from ingested algal food by slug digestive gland cells. Thylakoid stacks and stroma of chloroplasts were engulfed by the slug cells. In the slug cells thylakoids were surrounded by one membrane only. This membrane is interpreted as having been generated by the mollusk during phagocytosis. It is inferred to be eukaryotic in origin and unlikely, therefore, to be endowed with the translocons system ordinarily regulating import of algal gene-encoded plastid preproteins. Our structural findings suggest that chloroplast components in the slug cells are thylakoid stacks with chloroplast stroma only.     

红豆草根瘤侵染细胞核在细胞凋亡中的超微结构变化

用透射电镜观察红豆草根瘤侵染细胞核在细胞凋亡过程中的超微结构,以探讨红豆草根瘤侵染细胞核在发育过程中的超微结构变化及其与细胞凋亡的关系.结果表明,红豆草根瘤侵染细胞核的超微结构随细胞发育程度不同而不同.在幼龄侵染细胞中,细胞核体积较大,近似圆形.在即将成熟和成熟的侵染细胞中,细胞核膜有内陷现象,其核仁常具有核仁泡和核仁联合体.在早期凋亡的侵染细胞中,细胞核体积减小,形状变得不规则,核膜出现大量内陷,在其表面形成许多大的突起和深的沟槽,有时还有内质网、线粒体、小液泡和细菌等位于核膜的内陷处,而且核仁也开始裂解.在后期凋亡的侵染细胞中,除细菌解体外,还出现核仁消失,核膜破裂,核质外流,并在细胞质中形成一些电子密度很高,无一定形状的团块状物质.     

Aerial migration of the Dictyostelium slug

The Dictyostelium slug lays down curved marks in its slime sheath trail as it migrates across an agar substrate. These 'footprints' are caused by elevation of the slug anterior as it initiates a period of aerial migration and can be used as a measure of the slug's propensity for this behavior. A variety of factors have been found to affect the number of footprints created per distance migrated. Smaller slugs produce a higher incidence of footprints than larger slugs. Migration in the light and lower temperatures during migration increase footprint incidence. Activated charcoal reduces, while exogenous addition of ammonia increases, the incidence of footprints. Simulation of the three-dimensional (3D) environment of the soil suggests that aerial migration plays a role in the slug's movement through the cavities of its natural environment. A model proposes that aerial migration is initiated by a small group of continually changing prestalk cells that acts as a pacemaker and is moved around the circumference of the slug tip by the rotation of the prestalk cells. As this pacemaker reaches the upper surface of the slug it can initiate aerial migration.     

Dynamic alterations in gene expression after Wnt-mediated induction of avian neural crest

The Wnt signaling pathway is important in the formation of neural crest cells in many vertebrates, but the downstream targets of neural crest induction by Wnt are largely unknown. Here, we examined quantitative changes in gene expression regulated by Wnt-mediated neural crest induction using quantitative PCR (QPCR). Induction was recapitulated in vitro by adding soluble Wnt to intermediate neural plate tissue cultured in collagen, and induced versus control tissue were assayed using gene-specific primers at times corresponding to premigratory (18 and 24 h) or early (36 h) stages of crest migration. The results show that Wnt signaling up-regulates in a distinct temporal pattern the expression of several genes normally expressed in the dorsal neural tube (slug, Pax3, Msx1, FoxD3, cadherin 6B) at "premigratory" stages. While slug is maintained in early migrating crest cells, Pax3, FoxD3, Msx1 and cadherin 6B all are down-regulated by the start of migration. These results differ from the temporal profile of these genes in response to the addition of recombinant BMP4, where gene expression seems to be maintained. Interestingly, expression of rhoB is unchanged or even decreased in response to Wnt-mediated induction at all times examined, though it is up-regulated by BMP signals. The temporal QPCR profiles in our culture paradigm approximate in vivo expression patterns of these genes before neural crest migration, and are consistent with Wnt being an initial neural crest inducer with additional signals like BMP and other factors maintaining expression of these genes in vivo. Our results are the first to quantitatively describe changes in gene expression in response to a Wnt or BMP signal during transformation of a neural tube cell into a migratory neural crest cell.     

REPOPULATION OF POSTMITOTIC NUCLEOLI BY PREFORMED RNA : II. Ultrastructure

The reconstruction of the nucleolus after mitosis was analyzed by electron microscopy in cultured mammalian (L929) cells in which nucleolar RNA synthesis was inhibited for a 3 h period either after or before mitosis. When synchronized mitotic cells were plated into a concentration of actinomycin D sufficient to block nucleolar RNA synthesis preferentially, nucleoli were formed at telophase as usual. 3 h after mitosis, these nucleoli had fibrillar and particulate components and possessed the segregated appearance characteristic of nucleoli of actinomycin D-treated cells. Cells in which actinomycin D was present for the last 3 h preceding mitosis did not form nucleoli by 3 h after mitosis though small fibrillar prenucleolar bodies were detectable at this time. These bodies subsequently grew in size and eventually acquired a particulate component. It took about a full cell cycle before nucleoli of these cells were completely normal in appearance. Thus, nucleolar RNA synthesis after mitosis is not necessary for organization of nucleoli after mitosis. However, inhibition of nucleolar RNA synthesis before mitosis renders the cell incapable of forming nucleoli immediately after mitosis. If cells are permitted to resume RNA synthesis after mitosis, they eventually regain nucleoli of normal morphology.     

Persistent Nucleoli in Early Postimplantation of Rat Embryos

Ultrastructural changes of the nucleolus in mitotic embryonic ectodermal cells of 7 1/2-day and 7 2/3-day rat embryos were examined. It was found that the nucleolus was broken down into small fragments during late prophase and metaphase, and that some of these fragments persisted in the cytoplasm of telophase cell (persistent nucleoli). No interphase embryonic ectodermal cells contained persistent nucleoli. Persistent nucleoli were also found in telophase cells of extraembryonic ectoderm, extraembryonic visceral endoderm and parietal endoderm of the embryos, but they disappeared in interphase cells. Persistent nucleoli in telophase cells tended to decrease in size with embryonic age, and they had almost completely disappeared in neuroectodermal cells of the telencephalon in 14 1/2-day embryos. They were concluded to be remnants of disappearing nucleoli in embryonic cells that were cycling too rapidly to permit their nucleoli to disappear completely.     

Nuclear and mitochondrial alterations in amebae exposed to ethidium bromide

Cultures of Amoeba proteus were exposed to ethidium bromide at concentrations ranging between 5 and 100 μg/ml for periods of up to 1 week. Samples of treated and control cells were prepared at intervals for electron microscopy. The main ultrastructural alterations were in nucleoli and in mitochondria. The nucleoli of treated cells increased in density and became spherical with more sharply defined margins than those of normal amebae. Many nucleoli contained electron-lucent regions or nucleolar vacuoles of varying size. The chromatin was unusually condensed in some amebae. Mitochondria developed a central electron-lucent region and accumulated a dense material in the matrix. Some cristae were abnormally dilated. The nuclear alterations occurred at least as early as the mitochondrial changes and were present even in cells exposed to the lower concentrations of inhibitor, in which mitochondrial changes were minimal.     

Concerted Movement of Prestalk Cells in Migrating Slugs of Dictyostelium Revealed by the Localization of Myosin

Localization of myosin in slugs of the cellular slime mold Dictyostelium discoideum was investigated by an immunofluorescence technique. Myosin is thought to provide the molecular machinery for cellular movement. We found that myosin could be visualized as c-shaped fluorescence at the cortex of prestalk cells in a migrating slug, and that the open regions of all c-shaped fluorescence point in the direction of the slug's migration. We reported previously that the c-shaped fluorescence of myosin can be seen at the cortex of the tail region of actively locomoting cells at the unicellular stage (39, 41). These results suggest that prestalk cells move actively in the slug, and that their direction of movement, which can be identified from the polarity of c-shaped fluorescence, correspond with the direction of the slug's migration. The distribution of c-shaped fluorescence in slugs during migration, phototaxis and avoidance of ammonia strongly suggests that the slug's behavior is controlled by the concerted movement of prestalk cells.     

Cyclic-AMP binding to the cell surface during development of Dictyostelium discoideum

Abstract. Cell aggregation in Dictyostelium discoideum is a chemotactic process mediated by cyclic adenosine monophosphate (CAMP), which is detected by cell surface receptors. The cAMP signal is degraded by cAMP phosphodiesterase. The possibility that cAMP signals are also used for cell communication in the multicellular stages was studied by determining whether the cAMP receptors, which are essential for signal transduction, continue to function in these stages. During slug migration, the number of binding sites per cell decreases to about 15% of the maximum level acquired during aggregation. At the onset of fruiting body formation, a three- to Four-Fold increase in cAMP binding activity occurs. This increase coincides with an increase in cAMP phosphodiesterase. Both phenomena suggest that cell-cell communication mediated by cAMP is used during culmination. During both slug migration and early culmination, the prestalk cells exhibit about twice as much binding activity as the prespore cells.     

Orientation of cells during slug formation inDictyostelium

Summary Scanning electron microscopic observations ofDictyostelium discoideum cell masses during slug formation revealed two populations around the anterior tip; one group of cells resembled elongated aggregation stream cells and their orientation suggested that they move to the tip, whereas the other group of cells were isodiametric and showed no obvious orientation. In seeking further evidence for a role of differential cAMP chemotaxis in the orientation and movement of slug cells the anterior prestalk cells were compared to the posterior prespore cells in two chemotaxis tests. When a cell mass is placed on cAMP agar the prestalk cells exhibited better movement to cAMP sources but when the gradient was generated in a diffusion chamber the prestalk cells did not. This evidence suggested that the cells which are better able to generate a cAMP gradient might form part of the anterior zone of the slug.     

Function of ammonium transporter A in the initiation of culmination of development in Dictyostelium discoideum

The histidine kinase DhkC controls a phosphorelay involved in regulating the slug versus culmination choice during the multicellular developmental program of Dictyostelium discoideum. When the relay is active, slug migration is favored due to the activation of a cyclic AMP (cAMP) phosphodiesterase and the resultant lowering of the intracellular and extracellular levels of cAMP. Ammonia signaling represents one input into the DhkC phosphorelay, and previous studies indicated that the ammonium transporter C inhibits the relay in response to low ammonia levels. Evidence is presented that another member of the family of ammonium transporters, AmtA, also regulates the slug/culmination choice. Under standard conditions of development, the wild-type strain requires a transitional period of 2 to 3 h to go from fingers to culminants, with some slugs forming and migrating briefly prior to culmination. In contrast, amtA null cells, like cells that lack DhkC, possessed a transitional period of only 1 to 2 h and rarely formed slugs. Disruption of amtA in an amtC null strain overcame the slugger phenotype of that strain and restored its ability to culminate. Strains lacking AmtA were insensitive to the ability of ammonia to promote and prolong slug migration. These findings lead to the proposal that AmtA functions in ammonia sensing as an activator of the DhkC phosphorelay in response to perceived high ammonia levels.     

Nucleoli and ploidy in Potorous cells (PTK2) in vitro

Most cells of the male PTK₂ cell line contain one nucleolus, and they are diploid. However, a proportion of cells have more than one nucleolus. Cells with two and three nucleoli were isolated and cloned into viable populations. Greater than 90% of the cells in these clonal populations maintained the abnormal nucleolar number of the originally isolated cell. Karyotype analysis of cells with two and three nucleoli demonstrated that the cells were respectively tetraploid and hexaploid. It was concluded that in PTK₂ cells, nucleolarity is a good index of ploidy even if the ploidy level is abnormal. Furthermore, long term monitoring of the tetraploid cells demonstrated virtually no tendency towards reversion to the diploid condition as suggested by other studies in Potorous.