Dcas Supports Cell Polarization and Cell-Cell Adhesion Complexes in Development

Mammalian Cas proteins regulate cell migration, division and survival, and are often deregulated in cancer. However, the presence of four paralogous Cas family members in mammals (BCAR1/p130Cas, EFS/Sin1, NEDD9/HEF1/Cas-L, and CASS4/HEPL) has limited their analysis in development. We deleted the single Drosophila Cas gene, Dcas, to probe the developmental function of Dcas. Loss of Dcas had limited effect on embryonal development. However, we found that Dcas is an important modulator of the severity of the developmental phenotypes of mutations affecting integrins (If and mew) and their downstream effectors Fak56D or Src42A. Strikingly, embryonic lethal Fak56D-Dcas double mutant embryos had extensive cell polarity defects, including mislocalization and reduced expression of E-cadherin. Further genetic analysis established that loss of Dcas modified the embryonal lethal phenotypes of embryos with mutations in E-cadherin (Shg) or its signaling partners p120- and β-catenin (Arm). These results support an important role for Cas proteins in cell-cell adhesion signaling in development.     

Cell Division in Tetraspora

Cell division in Tetraspora sp. is described. The cell becomesimmotile some while before mitosis and the basal bodies withdrawfrom the cell surface. The preprophase nucleus migrates to thebasal body complex, around which increasing numbers of microtubulesgather. The spindle is closed with open polar fenestrae; a basalbody complex is always closely associated with at least onepole. No spindles were observed to have basal bodies at bothpoles, and the spindle may possibly be unicentric. During anaphase,spindle microtubules penetrate through the fenestrae. Aftertelophase, the nuclei come together as a phycoplast forms betweenthem; cytokinesis is effected by furrowing. Forming basal bodiesare frequently encountered in late telophase and cleaving cells;no evidence was obtained that the basal bodies replicated beforemitosis. The protoplast rotates inside the cell wall duringcleavage. Cell division is compared with that of other greenalgae, and in particular, Chlamydomonas.     

Cell Division in Tetraedron

Mitosis and cytokinesis in Tetraedron are described. Persistentcentrioles replicate before division and the pairs separateto define the future poles of the spindle whilst increasingnumbers of microtubules become associated with them. By prophase,the centrioles and most extranuclear microtubules have becomeenclosed within a 'perinuclear envelope' of endoplasmic reticulum.The nuclear envelope near the centrioles then becomes indentedand finally ruptures to form polar fenestrae during prometaphase;the extranuclear microtubules soon vanish and appear to movethrough the fenestrae into the forming spindle. Metaphase, anaphase,and telophase follow as usual. After mitosis, arrays of 'phycoplast'microtubules proliferate between nuclei. The cytoplasm is cleavedby membrane furrows coplanar with and growing through the phycoplasttubules. However, this cleavage is delayed until the cells havebecome multinucleate, and it appears to be irregular in extentand disposition in the cell until after a final set of synchronousmitoses. Then cytokinesis cuts up the cytoplasm into numeroussmall autospores which secrete their own wall; they are laterreleased following rupture of the parental wall. Some autosporesare binucleate which indicates that this cleavage apparatusdoes not necessarily cut up all the cytoplasm into uninucleatesegments. Vegetative reproduction in these organisms is comparedto that of other members of the Chlorococcales.     

Polarization of Drosophila Neuroblasts During Asymmetric Division

During Drosophila development, neuroblasts divide to generate progeny with two different fates. One daughter cell self-renews to maintain the neuroblast pool, whereas the other differentiates to populate the central nervous system. The difference in fate arises from the asymmetric distribution of proteins that specify either self-renewal or differentiation, which is brought about by their polarization into separate apical and basal cortical domains during mitosis. Neuroblast symmetry breaking is regulated by numerous proteins, many of which have only recently been discovered. The atypical protein kinase C (aPKC) is a broad regulator of polarity that localizes to the neuroblast apical cortical region and directs the polarization of the basal domain. Recent work suggests that polarity can be explained in large part by the mechanisms that restrict aPKC activity to the apical domain and those that couple asymmetric aPKC activity to the polarization of downstream factors. Polarized aPKC activity is created by a network of regulatory molecules, including Bazooka/Par-3, Cdc42, and the tumor suppressor Lgl, which represses basal recruitment. Direct phosphorylation by aPKC leads to cortical release of basal domain factors, preventing them from occupying the apical domain. In this framework, neuroblast polarity arises from a complex system that orchestrates robust aPKC polarity, which in turn polarizes substrates by coupling phosphorylation to cortical release.Cells use polarity for remarkably diverse functions. In this article, I discuss a polarity that is harnessed to generate daughter cells with different fates. Using polarity to divide asymmetrically addresses several challenges that complex organisms face. The diversification of cell types and tissues that occurs during the development of complex organisms is one such challenge. Drosophila neuroblasts, the subject of this article, undergo repeated symmetry breaking asymmetric cell divisions (ACDs) to populate the central nervous system. In a similar manner in adult organisms, ACDs are important for adult homeostasis, replenishing cells that are turned over during the course of normal physiology (Betschinger and Knoblich 2004).A fundamental aspect of ACD is the production of daughter cells containing distinct fate determinants. To segregate fate determinants, the cell becomes polarized to form mutually exclusive cortical domains, each with a set of fate determinants appropriate for one of the two daughter cells. The cleavage furrow forms at the interface of the two domains, partitioning the fate determinants into the two daughter cells where they function to either self-renew (to keep the progenitor population) or to differentiate (e.g., by changing the pattern of gene expression). One of the unique features of the symmetry breaking that occurs during ACD, at least as implemented by the neuroblast, is that it is remarkably dynamic, developing early in mitosis and depolarizing following the completion of cytokinesis.Since the discovery of the first polarized components, neuroblasts have been an excellent model system for investigating the mechanisms of cell polarization and have been extensively analyzed. Although aspects of neuroblast polarity remain unclear, a core framework for how polarity is created and maintained is emerging. In this article, I focus on neuroblast polarity as centered around the activity of atypical protein kinase C, which has emerged as a key regulator of the process. In this framework, neuroblast polarity can be explained by events that polarize aPKC and those that couple aPKC activity to the polarization of fate determinants.     

Tropomyosin in Preimplantation Mouse Development: Identification, Expression, and Organization during Cell Division and Polarization

Tropomyosin is an actin-binding cytoskeletal protein which has been extensively characterized in a variety of cell types and tissues, with the exception of very early developmental stages during which cellular polarization first occurs. We have identified five polypeptides in mouse preimplantation conceptuses which show many of the characteristics of tropomyosin. They form the major portion of the heat-stable cytoskeletal protein fraction of blastomeres and have the characteristic isoelectric and SDS–PAGE migration characteristics on 1-D and 2-D gels. All five polypeptides were synthesized in late 2- and 4-cell, and all 8-cell stages, with three of the five polypeptides showing lower synthetic levels in fertilized eggs and early 2-cell conceptuses. These heat-stable proteins showed specific differences from proteins isolated from mouse 3T3 fibroblasts by the same method, namely higherM_risoforms were not represented, also some of the isoforms can be labeled by incorporation of [¹⁴C]proline. The cellular distribution of tropomyosin in early stage conceptuses was examined using monoclonal and affinity-purified polyclonal antibodies. Tropomyosin becomes associated both with the blastomere cortex postfertilization and with the cleavage furrow during cytokinesis. The interphase cortical association is uniform until the 8-cell stage, when tropomyosin becomes associated with the developing apical pole and is excluded from the basolateral cortex. This polar localization is inherited along with the pole at the 8- to 16-cell division, but experiments in which cell division is artificially prolonged show that tropomyosin localization does not represent a permanent marking of the pole. We conclude that the early mouse conceptus contains a unique and specific set of tropomyosins which respond to polarizing signals.     

Polarity in Stem Cell Division: Asymmetric Stem Cell Division in Tissue Homeostasis

Many adult stem cells divide asymmetrically to balance self-renewal and differentiation, thereby maintaining tissue homeostasis. Asymmetric stem cell divisions depend on asymmetric cell architecture (i.e., cell polarity) within the cell and/or the cellular environment. In particular, as residents of the tissues they sustain, stem cells are inevitably placed in the context of the tissue architecture. Indeed, many stem cells are polarized within their microenvironment, or the stem cell niche, and their asymmetric division relies on their relationship with the microenvironment. Here, we review asymmetric stem cell divisions in the context of the stem cell niche with a focus on Drosophila germ line stem cells, where the nature of niche-dependent asymmetric stem cell division is well characterized.Asymmetric cell division allows stem cells to self-renew and produce another cell that undergoes differentiation, thus providing a simple method for tissue homeostasis. Stem cell self-renewal refers to the daughter(s) of stem cell division maintaining all stem cell characteristics, including proliferation capacity, maintenance of the undifferentiated state, and the capability to produce daughter cells that undergo differentiation. A failure to maintain the correct stem cell number has been speculated to lead to tumorigenesis/tissue hyperplasia via stem cell hyperproliferation or tissue degeneration/aging via a reduction in stem cell number or activity (Morrison and Kimble 2006; Rando 2006). This necessity changes during development. The stem cell pool requires expansion earlier in development, whereas maintenance is needed later to sustain tissue homeostasis.There are two major mechanisms to sustain a fixed number of adult stem cells: stem cell niche and asymmetric stem cell division, which are not mutually exclusive. Stem cell niche is a microenvironment in which stem cells reside, and provides essential signals required for stem cell identity (Fig. 1A). Physical limitation of niche “space” can therefore define stem cell number within a tissue. Within such a niche, many stem cells divide asymmetrically, giving rise to one stem cell and one differentiating cell, by placing one daughter inside and another outside of the niche, respectively (Fig. 1A). Nevertheless, some stem cells divide asymmetrically, apparently without the niche. For example, in Drosophila neuroblasts, cell-intrinsic fate determinants are polarized within a dividing cell, and subsequent partitioning of such fate determinants into daughter cells in an asymmetric manner results in asymmetric stem cell division (Fig. 1B) (see Fig. 3A and Prehoda 2009).Open in a separate windowFigure 1.Mechanisms of asymmetric stem cell division. (A) Asymmetric stem cell division by extrinsic fate determinants (i.e., the stem cell niche). The two daughters of stem cell division will be placed in distinct cellular environments either inside or outside the stem cell niche, leading to asymmetric fate choice. (B) Asymmetric stem cell division by intrinsic fate determinants. Fate determinants are polarized in the dividing stem cells, which are subsequently partitioned into two daughter cells unequally, thus making the division asymmetrical. Self-renewing (red line) and/or differentiation promoting (green line) factors may be involved.In this review, we focus primarily on asymmetric stem cell divisions in the Drosophila germ line as the most intensively studied examples of niche-dependent asymmetric stem cell division. We also discuss some examples of stem cell division outside Drosophila, where stem cells are known to divide asymmetrically or in a niche-dependent manner.     

Osmotic Adjustment and Stomatal Response to Water Deficits in Maize

A pot experiment was carried out using five maize {Zea maysL.) cultivars under three soil moisture levels (MPa 0 to –0.05,–0.3 to –0.9 and –1.2 to –1.5) to investigatethe effects of water deficits on osmotic adjustment and stomatalconductance. The degree of leaf rolling and the sugar and nutrientconcentrations in leaf cell sap were measured. Leaf water potential and osmotic potential decreased and stomatalconductance decreased with increasing water deficits. Stomatalconductance correlated positively with leaf water potentialand osmotic potential. Degree of leaf rolling was lower in cultivarswhich maintained higher turgor. Osmotic adjustment of 0.08 to0.43 MPa was found under the lowest soil moisture level in fivecultivars used. Sugar and K were the major osmotic substancesin the maize plant. Sugar, K and Mg concentrations increasedunder water deficit, and correlated negatively with a decreasein osmotic potential. Key words: Zea mays L., leaf water relations, leaf rolling, osmotic adjustment, stomatal conductance, water deficit     

细胞不对称分裂

细胞不对称分裂是多细胞生物发育的基础。细胞不对称分裂的重要特征是细胞命运决定子在细胞分裂期间的不对称分离。细胞不对称分裂一般要经历4个步骤：在细胞中建立一个极性轴;沿此轴定向并形成纺锤体;细胞命运决定子沿极性轴作极性分布;细胞分裂后,不同的细胞命运决定子指导决定细胞的不同命运。     

Localization of the Functional p34cdc2 Homolog of Maize in Root Tip and Stomatal Complex Cells: Association with Predicted Division Sites

We have used an antibody against the functional homolog of the cdc2 kinase of maize to localize the p34cdc2 protein within dividing cells of the root apex and the stomatal complex of leaf epidermis. The microtubule cytoskeletal structure of plant cells was visualized concomitantly with a monoclonal antibody specific for [alpha]-tubulin. We found that the cdc2 protein is localized mainly to the nucleus in plant cells at interphase and early prophase. This finding contrasts markedly with the predominantly cytoplasmic staining obtained using antibody to the PSTAIRE motif, which is common to cdc2 and numerous cdc2-like proteins. In a subpopulation of root cells at early prophase, the p34cdc2 protein is also distributed in a band bisecting the nucleus. Double labeling with the maize p34cdc2Zm antibody and tubulin antibody revealed that this band colocalizes with the preprophase band (PPB) of microtubules, which predicts the future division site. Root cells in which microtubules had been disrupted with oryzalin did not contain this band of p34cdc2 protein, suggesting that formation of the microtubule PPB is necessary for localization of the p34cdc2 kinase to the plane of the PPB. The p34cdc2 protein is also localized to the nucleus and PPB in cells that give rise to the stomatal complex, including those cells preparing for the highly asymmetrical divisions that produce subsidiary cells. Association of the p34cdc2 protein with the PPB suggests that the cdc2 kinase has a role in establishing the division site of plant cells and, therefore, a role in plant morphogenesis.     

Regulation of Cell Division in Arabidopsis

The study of cell cycle control in plants is expected to contribute to the understanding of plants' unique developmental features. The principal regulators of the eukaryotic cell cycle, namely, cyclin-dependent kinases (CDKs) and cyclins, are also conserved in plants. This review is concerned with our present knowledge on cell cycle regulation in Arabidopsis thaliana, which is widely accepted as a model plant for the study of a broad range of biological questions. Up to the present, 2 CDKs and 11 cyclins have been identified in Arabidopsis. While the expression of one of these CDKs has been found to be positively correlated with the competence of cells to divide, cyc1A1 expression of the cyclin has been almost exclusively confined to dividing cells. Although much remains to be studied concerning upstream regulators of these genes, the successful introduction of mutant CDKs into plants demonstrates the potential of using such an approach to intentionally modulate the plant cell cycle and development.     

Outward-Rectifying K+ Channels in Stomatal Guard Cell Protoplasts

Ion channels in stomatal guard cell protoplasts from Vicia fabawere examined using the patch-clamp technique. Most ion channelshaving unit conductance ranging between 10 and 30 pS showedclear outward-rectification in symmetrical 50mM KCl. The largeinside-out membranes contained these outward-rectifiers as themajor and relatively stable channels. The channels were K⁺ ion-selective.Kinetic analysis revealed that the channels have three conductancestates: open, closed and inactivated. The rates of transitionto and from the inactivated state were highly voltage-dependent. (Received April 6, 1988; Accepted May 25, 1988)     

Determination of Fluorescence Polarization of Double Chromophore Complexes

Polarized fluorescence of rigid double-chromophore complexes with intracomplex energy exchange between chromophores was analyzed, and the formula for the degree of polarization derived for the case of steady-state excitation: P = (3 cos²θ - 1 + 2A)/(3 + cos²θ + 4A). In this formula θ is the angle between the transition dipole moments of chromophores in complexes, and A is the parameter dependent on the spectroscopic features of chromophores and energy migration rates. The case of excitation by a δ-pulse was also analyzed, and a formula for fluorescence polarization kinetics was derived.As an example of the application of the derived formulae, the polarized fluorescence spectra and their picosecond kinetics were calculated for the β-subunits of the blue-green algae Agmenellum quadruplicatum. The results obtained were compared with experimental measurements of Mimuro et al. (1986, Biochim. Biophys. Acta848, 155-166) and found to match these data well.     

存活蛋白与细胞分裂

凋亡抑制蛋白家族的新成员存活蛋白(survivin)在肿瘤细胞中特异性地表达,它不仅具有抗凋亡的功能,而且它在细胞分裂中的重要作用也越来越受到人们关注,这一方面的研究取得了新的进展。深入研究存活蛋白在细胞分裂中的作用,将拓展人们对于这一分子新的认识。     

B细胞不对称分裂

细胞的不对称分裂对于细胞多样性产生的重要性已经被大部分人所认识。B细胞的不对称分裂首先是在抗体类别转换的研究中发现的。最近,美国5科学家对B细胞在免疫发生中心中不对称分裂的原因进行了探索。结果发表在2012年1月20日出版的《Science》中。B细胞的不对称分裂参与体液免疫的抗体类别转换和抗体亲和力成熟过程。对于其机制仍不清楚,但目前研究初步提示细胞内分子的不对称分布是其发生的上游因素。并且B细胞的不对称分裂可能与不对称抗原分离可能在抗体亲和力成熟过程中具有独立协同作用。