Determining cell shape: adaptive regulation of cyanobacterial cellular differentiation and morphology

Similar to other bacteria, cyanobacteria exist in a wide-ranging diversity of shapes and sizes. However, three general shapes are observed most frequently: spherical, rod and spiral. Bacteria can also grow as filaments of cells. Some filamentous cyanobacteria have differentiated cell types that exhibit distinct morphologies: motile hormogonia, nitrogen-fixing heterocysts, and spore-like akinetes. Cyanobacterial cell shapes, which are largely controlled by the cell wall, can be regulated by developmental and/or environmental cues, although the mechanisms of regulation and the selective advantage(s) of regulating cellular shape are still being elucidated. In this review, recent insights into developmental and environmental regulation of cell shape in cyanobacteria and the relationship(s) of cell shape and differentiation to organismal fitness are discussed.     

Influence of changes in sea water salinity on the growth,photosynthetic pigment content,and cell size of the benthic alga <Emphasis Type="Italic">Attheya ussurensis</Emphasis> Stonik,Orlova et Crawford, 2006 (Bacillariophyta)

The study deals with the effect of changes in salinity from 32 to 4‰ (at an interval of 4‰) on the growth, chlorophyll a and carotenoid contents, and cell size of the benthic alga Attheya ussurensis (Bacillariophyta). A. ussurensis showed high tolerance to reduced salinity and ability to adapt to salinity changes from 16 to 12‰. In this salinity range, the cells restored their shapes, sizes, and physiological functions. The number of cells and photosynthetic pigment content were highest at a salinity reduction to 24‰. At 8‰, algal cells remained alive, but the process of cell division was inhibited; as a result, the number of cells was significantly lower than in the control, the cells did not restore their sizes and shapes and remained deformed until the end of the experiment. A drop in salinity to 4‰ caused a complete loss of cell viability of A. ussurensis within a day of exposure to this factor.     

Growth regulation in the mycobacterial cell

A framework was developed to provide an integrated view of mycobacterial growth and its regulation. The topics reviewed include the properties of cell cultures and their relation to properties of individual cells, cell sizes and macromolecular compositions, uptake of nutrients through the cell envelope, protein biosynthesis, core metabolic pathways, generation of an electrochemical gradient of protons, ATP synthesis and the control of energy generation.     

Bacterial cell shape

Bacterial species have long been classified on the basis of their characteristic cell shapes. Despite intensive research, the molecular mechanisms underlying the generation and maintenance of bacterial cell shape remain largely unresolved. The field has recently taken an important step forward with the discovery that eukaryotic cytoskeletal proteins have homologues in bacteria that affect cell shape. Here, we discuss how a bacterium gains and maintains its shape, the challenges still confronting us and emerging strategies for answering difficult questions in this rapidly evolving field.     

Cell wall and cell growth characteristics of giant‐celled algae

Because of their large sizes and simple shapes, giant‐celled algae have been used to study how the structural and mechanical properties of cell walls influence cell growth. Here we review known relationships between cell wall and cell growth properties that are characteristic of three representative taxa of giant‐celled algae, namely, Valonia ventricosa, internodal cells of characean algae, and Vaucheria frigida. Tip‐growing cells of the genus Vaucheria differ from cells undergoing diffuse growth in V. ventricosa and characean algae in terms of their basic architectures (non‐lamellate vs. multilamellate) and their dependence upon pH and Ca²⁺ for cell wall extensibility. To further understand the mechanisms controlling cell growth by cell walls, comparative analyses of cell wall structures and/or associated growth modes will be useful. The giant‐celled algae potentially serve as good models for such investigations because of their wide variety of developmental processes and cell shapes exhibited.     

Morphometry of normal, regenerating and cancerous hepatocytes

During the morphometric analysis of liver cell carcinomas arising in cirrhotic livers, the sizes and shapes of 2200 cancerous and 1800 regenerative hepatocytes were measured and compared to normal hepatocytes. The neoplastic population showed significantly higher polymorphism, nucleocytoplasmic ratio and the percentage of multinucleated cells, whereas the sizes of cancerous cells were the smallest. Values for regenerative cells were mainly between those of neoplastic and normal cells. The exception was constituted by the group of very large regenerative cells which met the criteria of large liver cell dysplasia (LLCD). Low nucleocytoplasmic ratio achieved by these cells is consistent with the hypothesis of regenerative character of LLCD.     

Mechanisms of bacterial morphogenesis: Evolutionary cell biology approaches provide new insights

How Darwin's “endless forms most beautiful” have evolved remains one of the most exciting questions in biology. The significant variety of bacterial shapes is most likely due to the specific advantages they confer with respect to the diverse environments they occupy. While our understanding of the mechanisms generating relatively simple shapes has improved tremendously in the last few years, the molecular mechanisms underlying the generation of complex shapes and the evolution of shape diversity are largely unknown. The emerging field of bacterial evolutionary cell biology provides a novel strategy to answer this question in a comparative phylogenetic framework. This relatively novel approach provides hypotheses and insights into cell biological mechanisms, such as morphogenesis, and their evolution that would have been difficult to obtain by studying only model organisms. We discuss the necessary steps, challenges, and impact of integrating “evolutionary thinking” into bacterial cell biology in the genomic era.     

The 14-3-3 proteins of Trypanosoma brucei function in motility, cytokinesis, and cell cycle

The cDNAs for two isoforms (I and II) of the 14-3-3 proteins have been cloned and functionally characterized in Trypanosoma brucei. The amino acid sequences of isoforms I and II have 47 and 50% identity to the human tau isoform, respectively, with important conserved features including a potential amphipathic groove for the binding of phosphoserine/phosphothreonine-containing motifs and a nuclear export signal-like domain. Both isoforms are abundantly expressed at approximately equal levels (1-2 x 10(6) molecules/cell) and localized mainly in the cytoplasm. Knockdown by induction of double-stranded RNA of isoform I and/or II in both bloodstream and procyclic forms resulted first in a reduction of cell motility and then significant reduction in cell growth rates and morphological changes; the changes include aberrant numbers of organelles and abnormal shapes and sizes that mimic phenotypes produced by various cytokinesis inhibitors. Morphological and fluorescence-activated cell sorting analysis of the cell cycle suggested that isoforms I and II might play important roles in nuclear (G2-M transition) and cell (M-G1 transition) division. These findings indicate that the 14-3-3 proteins play important roles in cell motility, cytokinesis, and the cell cycle.     

Fine structure of the cell surface and Golgi apparatus of Ochromonas

Ochromonas danica has an unusually flexible cell surface capable of producing projections of varying sizes and shapes: large projections, 340-360 nm long, and small projections, 50-110 nm long. These projections have been demonstrated by transmission and scanning electron microscopy; some of them may break off into the medium and be the source of extracellular membranes and vesicles reported in the cell-free O. danica growth medium. Ruthernium red stained the acid mucopolysaccharide layer just outside the cell surface as well as small blebs at the cell surface. The Golgi complex of O. danica, Ochromonas malhamensis, Ochromonas sociabilis and Ochromonas sp. produced small coated vesicles which may move toward and fuse with the plasma membrane. The role of the several vesicles is unknown but possible functions are discussed.     

Integrating cellular dimensions with cell differentiation during early development

Early embryo development is characterized by alteration of cellular dimensions and fating of blastomeres. An emerging concept is that cell size and shape drive cellular differentiation during early embryogenesis in a variety of model organisms. In this review, we summarize recent advances that elucidate the contribution of the physical dimensions of a cell to major embryonic transitions and cell fate specification in vivo. We also highlight techniques and newly evolving methods for manipulating the sizes and shapes of cells and whole embryos in situ and ex vivo. Finally, we provide an outlook for addressing fundamental questions in the field and more broadly uncovering how changes to cell size control decision making in a variety of biological contexts.     

The cell cycle,cell death,and cell morphology during retinoic acid-induced differentiation of embryonal carcinoma cells

Time-lapse films were made of PC13 embryonal carcinoma cells, synchronized by mitotic shake off, in the absence and presence of retinoic acid. Using a method based on the transition probability model, cell cycle parameters were determined during the first five generations following synchronization. In undifferentiated cells, cell cycle parameters remained identical for the first four generations, the generation time being 11–12 hr. In differentiating cells, with retinoic acid added at the beginning of the first cycle, the first two generations were the same as controls. The duration of the third generation, however, was increased to 15.7 hr while the fourth and fifth generation were approximately 20 hr, the same as in exponentially growing, fully differentiated cells. The increase in generation time of dividing cells was principally due to an increase in the length of S phase. Cell death induced by retinoic acid also occurred principally in the third and subsequent generations. Cell population growth was then significantly less than that expected from the generation time derived from cycle analysis of dividing cells. Cells lysed frequently as sister pairs suggesting susceptibility to retinoic acid toxicity determined in a generation prior to death. Morphological differentiation, as estimated by the area of substrate occupied by cells, was shown to begin in the second cell cycle after retinoic acid addition. These results demonstrate that as in the early mammalian embryo, differentiation of embryonal carcinoma cells to an endoderm-like cell is also accompanied by a decrease in growth rate but that this is preceded by acquisition of the morphology characteristic of the differentiated progeny.     

Variety of mitochondrial shapes,sizes, and volumes inChlamydomonas reinhardii

Summary Mitochondria in cells ofChlamydomonas reinhardii, which at an intermediate stage of the vegetative cell cycle have been submitted to gametogenesis under dark and cold conditions, remain more or less unchanged with and without the addition of chloramphenicol. They exist in various number, shapes, and sizes and can be branched or unbranched as well as small or large. Giant mitochondria can be fused to a mitochondrial network, which, in contrast to the previously reported network (Grobe andArnold 1975), lies predominantly in the center of the cell. Mitochondrial volumes were revealed by means of morphometrical analyses from serial sections of four entire cells.     

Joint modeling of cell and nuclear shape variation

Modeling cell shape variation is critical to our understanding of cell biology. Previous work has demonstrated the utility of nonrigid image registration methods for the construction of nonparametric nuclear shape models in which pairwise deformation distances are measured between all shapes and are embedded into a low-dimensional shape space. Using these methods, we explore the relationship between cell shape and nuclear shape. We find that these are frequently dependent on each other and use this as the motivation for the development of combined cell and nuclear shape space models, extending nonparametric cell representations to multiple-component three-dimensional cellular shapes and identifying modes of joint shape variation. We learn a first-order dynamics model to predict cell and nuclear shapes, given shapes at a previous time point. We use this to determine the effects of endogenous protein tags or drugs on the shape dynamics of cell lines and show that tagged C1QBP reduces the correlation between cell and nuclear shape. To reduce the computational cost of learning these models, we demonstrate the ability to reconstruct shape spaces using a fraction of computed pairwise distances. The open-source tools provide a powerful basis for future studies of the molecular basis of cell organization.     

Extramacrochaetae,a trans-acting gene of the achaete-scute complex of Drosophila involved in cell communication

Summary Phenotypic analyses of genetic combinations involving the gene extramacrochaetae (emc) reveal its participation in the differentiation of both sensory elements and wing veins. The study of near-amorphic alleles of emc in mitotitc recombination clones indicates that it also affects cell proliferation. These clones show abnormal sizes, shapes and spatial distribution. They differentiate extra sensory elements as well as extra veins. A gain of function mutation in the gene causes opposite phenotypes in both differentiation systems. The effects of the mutant on proliferation and patterning are consistent with the emc gene being involved in the transfer of information between neighbouring cells, which leads to the spatial expression of the achaetescute gene complex and genes involved in vein formation.     

Creation of arrays of cell aggregates in defined patterns for developmental biology studies using dielectrophoresis

It is shown that dielectrophoresis—the movement of particles in non‐uniform electric fields—can be used to create engineered skin with artificial placodes of different sizes and shapes, in different spatial patterns. Modeling of the electric field distribution and image analysis of the cell aggregates produced showed that the aggregation is highly predictable. The cells in the aggregates remain viable, and reorganization and compaction of the cells in the aggregates occurs when the artificial skin is subsequently cultured. The system developed could be of considerable use for the in vitro study of developmental processes where local variations in cell density and direct cell–cell contacts are important. Biotechnol. Bioeng. 2010;105: 945–954. © 2009 Wiley Periodicals, Inc.     

The kinetic significance of cell size : II. Size distributions of resting and proliferating cells during interphase

This second part in a two part report describes the kinetic, cell size and nuclear size characteristics of S phase cells and cells with greatly protracted generation times (‘resting’ cells) in a cell line of human lymphoid cells. The median cell and nuclear sizes of S phase cells were greater than the corresponding median sizes observed in the whole population. Resting cells (operationally defined as unlabelled cells after 5 days of continuous labelling with [³H]TdR) have cell and nuclear size distributions overlapping with the cell and nuclear size distributions of the whole population. These resting cells are kinetically characterized by means of the observed labelling index vs time data during continuous labelling. The implication of these results are discussed.     

Correlation between membrane potential, cell shape, and motile activity in Amoeba proteus

The various motile activities and cell shapes of Amoeba proteus grown in Chalkley's solution are correlated with definitive electrical membrane potentials. The same correlations were found when definitive motile activities and cell shapes were experimentally induced by changing the pH of the culture medium. The highest values of membrane potential (−70 mV) were measured in monopodial amebae during active locomotion. In resting cells, which prevail in acid or basic media, the membrane potential decreases to −5 mV. In those resting cells, which also stop internal cytoplasmic movement at basic pH, the membrane potential turns positive (+9 mV − +30 mV).     

The emergence of shape: notions from the study of the Drosophila tracheal system

The generation of bodies and body parts with specific shapes and sizes has been a longstanding issue in biology. Morphogenesis in general and organogenesis in particular are complex events that involve global changes in cell populations in terms of their proliferation, migration, differentiation and shape. Recent studies have begun to address how these synchronized changes are controlled by the genes that specify cell fate and by the ability of cells to respond to extracellular cues. In particular, a notable shift in this research has occurred owing to the ability to address these issues in the context of the whole organism. For such studies, the Drosophila tracheal system has proven to be a particularly appropriate model. Here, my aim is to highlight some ideas that have arisen through our studies, and those from other groups, of Drosophila tracheal development. Rather than providing an objective review of the features of tracheal development, I intend to discuss some selected notions that I think are relevant to the question of shape generation.     

Nucleolar size and the activity of rRNA transport in the course of morphogenetic reduction of cell dimensions

In antheridial filaments of Chara vulgaris during the first period of spermatogenesis which consists of 6 synchronous cell division cycles there occurs a gradual decrease in sizes of cells entering successive mitoses. Present studies indicate that this process is correlated with a considerable reduction of total nucleolar volume in late G2 phase which, in turn, brings about decrease in sizes of nucleoli reappearing in telophase of the subsequent cell cycle. A consequence of the above phenomenon evidenced using 3H uridine autoradiography is a gradual increase--from one generation to the next--in an amount of rRNA transported into cytoplasm due to an increase in number of small nucleoli which were found to be more active in transport than larger nucleoli. This process leads to a lowering of an increase in nucleolar volumes during consecutive interphase periods owing to a progressively limited accumulation of rRNA for the sake of daughter cells, i.e. to a spontaneously magnifying reduction of nucleolar sizes in the forthcoming cell generations. Thus, diminution of nucleoli observed during the development of antheridial filaments seems to be due to a chain-series of processes connected with mechanisms which possibly regulate rRNA transport and accompany morphogenetic events.     

Rotation and asymmetry of the mitotic spindle direct asymmetric cell division in the developing central nervous system

The asymmetric segregation of cell-fate determinants and the generation of daughter cells of different sizes rely on the correct orientation and position of the mitotic spindle. In the Drosophila embryo, the determinant Prospero is localized basally and is segregated equally to daughters of similar cell size during epidermal cell division. In contrast, during neuroblast division Prospero is segregated asymmetrically to the smaller daughter cell. This simple switch between symmetric and asymmetric segregation is achieved by changing the orientation of cell division: neural cells divide in a plane perpendicular to that of epidermoblast division. Here, by labelling mitotic spindles in living Drosophila embryos, we show that neuroblast spindles are initially formed in the same axis as epidermal cells, but rotate before cell division. We find that daughter cells of different sizes arise because the spindle itself becomes asymmetric at anaphase: apical microtubules elongate, basal microtubules shorten, and the midbody moves basally until it is positioned asymmetrically between the two spindle poles. This observation contradicts the widely held hypothesis that the cleavage furrow is always placed midway between the two centrosomes.