scholarly journals E-cadherin and LGN align epithelial cell divisions with tissue tension independently of cell shape

2017 ◽  
Vol 114 (29) ◽  
pp. E5845-E5853 ◽  
Author(s):  
Kevin C. Hart ◽  
Jiongyi Tan ◽  
Kathleen A. Siemers ◽  
Joo Yong Sim ◽  
Beth L. Pruitt ◽  
...  
Keyword(s):  
Cell Division ◽  
Cell Monolayer ◽  
Spindle Orientation ◽  
Tissue Cell ◽  
Cellular Behavior ◽  
Cell Monolayers ◽  
Daughter Cells ◽  
Cell Divisions ◽  
Tensile Forces ◽  
E Cadherin

Tissue morphogenesis requires the coordinated regulation of cellular behavior, which includes the orientation of cell division that defines the position of daughter cells in the tissue. Cell division orientation is instructed by biochemical and mechanical signals from the local tissue environment, but how those signals control mitotic spindle orientation is not fully understood. Here, we tested how mechanical tension across an epithelial monolayer is sensed to orient cell divisions. Tension across Madin–Darby canine kidney cell monolayers was increased by a low level of uniaxial stretch, which oriented cell divisions with the stretch axis irrespective of the orientation of the cell long axis. We demonstrate that stretch-induced division orientation required mechanotransduction through E-cadherin cell–cell adhesions. Increased tension on the E-cadherin complex promoted the junctional recruitment of the protein LGN, a core component of the spindle orientation machinery that binds the cytosolic tail of E-cadherin. Consequently, uniaxial stretch triggered a polarized cortical distribution of LGN. Selective disruption of trans engagement of E-cadherin in an otherwise cohesive cell monolayer, or loss of LGN expression, resulted in randomly oriented cell divisions in the presence of uniaxial stretch. Our findings indicate that E-cadherin plays a key role in sensing polarized tensile forces across the tissue and transducing this information to the spindle orientation machinery to align cell divisions.

Stimulated Virus Production in Vinblastine and Cytochalasin-Induced Multinucleate Cells

1970 ◽  
Vol 28 ◽  
pp. 186-187
Author(s):  
Krishan Awtar
Keyword(s):  
Cell Division ◽  
Normal Cell ◽  
Virus Production ◽  
Multinucleate Cells ◽  
Daughter Cells ◽  
Cell Divisions ◽  
Nuclear Membranes ◽  
Division 2 ◽  
Vinblastine Sulfate

Exposure of cells to low sublethal but mitosis-arresting doses of vinblastine sulfate (Velban) results in the initial arrest of cells in mitosis followed by their subsequent return to an “interphase“-like stage. A large number of these cells reform their nuclear membranes and form large multimicronucleated cells, some containing as many as 25 or more micronuclei (1). Formation of large multinucleate cells is also caused by cytochalasin, by causing the fusion of daughter cells at the end of an otherwise .normal cell division (2). By the repetition of this process through subsequent cell divisions, large cells with 6 or more nuclei are formed.

Unequal cleavage in leech embryos: zygotic transcription is required for correct spindle orientation in subset of early blastomeres

Development ◽  
1996 ◽  
Vol 122 (2) ◽  
pp. 599-606
Author(s):  
S.T. Bissen ◽  
C.M. Smith
Keyword(s):  
Cell Division ◽  
Early Period ◽  
Spindle Orientation ◽  
Mitotic Spindles ◽  
Maternal Control ◽  
Transcription Analysis ◽  
Cell Divisions ◽  
Zygotic Transcription ◽  
Different Types ◽  
Alpha Amanitin

Leech embryos undergo invariant sequences of equal and unequal cell divisions to give rise to identifiable progeny cells. While many of the early cleavages are under maternal control, the divisions of a subset of early blastomeres (the large cells of the D' lineage) are perturbed after the inhibition of zygotic transcription. Analysis of the different types of cells produced in embryos injected with the transcriptional inhibitor, alpha-amanitin, revealed that the symmetry of cell division is perturbed in these large D'-derived cells during this early period of development. These cells, which would normally undergo a series of equal and unequal cleavages, always undergo equal cleavages after the inhibition of zygotic transcription. It appears that zygotically transcribed gene product(s) are required in the large cells of the D' lineage to orient the mitotic spindles properly for these unequal cell cleavages.

scholarly journals A Scribble-E-cadherin complex controls daughter cell patterning by multiple mechanisms

2021 ◽  
Author(s):  
Anchi S Chann ◽  
Ye Chen ◽  
Tanja Kinwel ◽  
Patrick O Humbert ◽  
Sarah Russell
Keyword(s):  
Cell Division ◽  
Mitotic Spindle ◽  
Cell Patterning ◽  
Cell Junction ◽  
Multiple Cues ◽  
Daughter Cells ◽  
Control Orientation ◽  
Mcf10a Cells ◽  
And Control ◽  
E Cadherin

The fate of the two daughter cells is intimately connected to their positioning, which is in turn regulated by cell junction remodelling and orientation of the mitotic spindle. How multiple cues are integrated to dictate the ultimate patterning of daughters is not clear. Here, we identify novel mechanisms of regulation of daughter positioning in single MCF10A cells. The polarity protein, Scribble, links E-cadherin to NuMA and Arp2/3 signalling for sequential roles in daughter positioning. First Scribble transmits cues from E-cadherin localised in retraction fibres to control orientation of the mitotic spindle. Second, Scribble re-locates to the junction between the two daughters to allow a new E-cadherin-based-interface to form between them, influencing the width of the nascent daughter-daughter junction, generation of filopodia and subsequent cell patterning. Thus, E-cadherin and Scribble dynamically relocate to different intracellular sites during cell division to orient the mitotic spindle and control placement of the daughter cells after cell division.

scholarly journals Epithelial polarity and spindle orientation: intersecting pathways

2013 ◽  
Vol 368 (1629) ◽  
pp. 20130291 ◽  
Author(s):  
Dan T. Bergstralh ◽  
Timm Haack ◽  
Daniel St Johnston
Keyword(s):  
Epithelial Cells ◽  
Cell Fate ◽  
Spindle Orientation ◽  
Mitotic Spindles ◽  
Daughter Cells ◽  
Cell Divisions ◽  
Discs Large ◽  
Stem Cell Divisions ◽  
Fate Determinants ◽  
Cell Fate Determinants

During asymmetric stem cell divisions, the mitotic spindle must be correctly oriented and positioned with respect to the axis of cell polarity to ensure that cell fate determinants are appropriately segregated into only one daughter cell. By contrast, epithelial cells divide symmetrically and orient their mitotic spindles perpendicular to the main apical–basal polarity axis, so that both daughter cells remain within the epithelium. Work in the past 20 years has defined a core ternary complex consisting of Pins, Mud and Gαi that participates in spindle orientation in both asymmetric and symmetric divisions. As additional factors that interact with this complex continue to be identified, a theme has emerged: there is substantial overlap between the mechanisms that orient the spindle and those that establish and maintain apical–basal polarity in epithelial cells. In this review, we examine several factors implicated in both processes, namely Canoe, Bazooka, aPKC and Discs large, and consider the implications of this work on how the spindle is oriented during epithelial cell divisions.

scholarly journals Spindle orientation and epidermal morphogenesis

2013 ◽  
Vol 368 (1629) ◽  
pp. 20130016 ◽  
Author(s):  
Anita Kulukian ◽  
Elaine Fuchs
Keyword(s):  
Stem Cells ◽  
Basement Membrane ◽  
Cell Fate ◽  
Mitotic Spindle ◽  
Spindle Orientation ◽  
Asymmetric Cell Divisions ◽  
Daughter Cells ◽  
Cell Divisions ◽  
Molecular Machinery ◽  
Asymmetric Partitioning

Asymmetric cell divisions (ACDs) result in two unequal daughter cells and are a hallmark of stem cells. ACDs can be achieved either by asymmetric partitioning of proteins and organelles or by asymmetric cell fate acquisition due to the microenvironment in which the daughters are placed. Increasing evidence suggests that in the mammalian epidermis, both of these processes occur. During embryonic epidermal development, changes occur in the orientation of the mitotic spindle in relation to the underlying basement membrane. These changes are guided by conserved molecular machinery that is operative in lower eukaryotes and dictates asymmetric partitioning of proteins during cell divisions. That said, the shift in spindle alignment also determines whether a division will be parallel or perpendicular to the basement membrane, and this in turn provides a differential microenvironment for the resulting daughter cells. Here, we review how oriented divisions of progenitors contribute to the development and stratification of the epidermis.

scholarly journals Activation of Discs large by aPKC aligns the mitotic spindle to the polarity axis during asymmetric cell division

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Ognjen Golub ◽  
Brett Wee ◽  
Rhonda A Newman ◽  
Nicole M Paterson ◽  
Kenneth E Prehoda
Keyword(s):  
Cell Division ◽  
Mitotic Spindle ◽  
Asymmetric Cell Division ◽  
Intramolecular Interaction ◽  
Spindle Orientation ◽  
Orientation Factor ◽  
Daughter Cells ◽  
Kinase C ◽  
Discs Large ◽  
Fate Determinants

Asymmetric division generates cellular diversity by producing daughter cells with different fates. In animals, the mitotic spindle aligns with Par complex polarized fate determinants, ensuring that fate determinant cortical domains are bisected by the cleavage furrow. Here, we investigate the mechanisms that couple spindle orientation to polarity during asymmetric cell division of Drosophila neuroblasts. We find that the tumor suppressor Discs large (Dlg) links the Par complex component atypical Protein Kinase C (aPKC) to the essential spindle orientation factor GukHolder (GukH). Dlg is autoinhibited by an intramolecular interaction between its SH3 and GK domains, preventing Dlg interaction with GukH at cortical sites lacking aPKC. When co-localized with aPKC, Dlg is phosphorylated in its SH3 domain which disrupts autoinhibition and allows GukH recruitment by the GK domain. Our work establishes a molecular connection between the polarity and spindle orientation machineries during asymmetric cell division.

scholarly journals RAS-mediated suppression of PAR3 and its effects on SCC initiation and tissue architecture occur independently of hyperplasia

2020 ◽  
Vol 133 (23) ◽  
pp. jcs249102
Author(s):  
Ji Ling ◽  
Maria Sckaff ◽  
Manisha Tiwari ◽  
Yifang Chen ◽  
Jingting Li ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Cell Division ◽  
Mapk Pathway ◽  
Temporal Analysis ◽  
Current Evidence ◽  
Spindle Orientation ◽  
Tissue Architecture ◽  
Spatial Cues ◽  
Human Squamous Cell Carcinoma ◽  
E Cadherin

ABSTRACTProper epithelial development and homeostasis depends on strict control of oriented cell division. Current evidence shows that this process is regulated by intrinsic polarity factors and external spatial cues. Owing to the lack of an appropriate model system that can recapitulate the architecture of the skin, deregulation of spindle orientation in human epithelial carcinoma has never been investigated. Here, using an inducible model of human squamous cell carcinoma (SCC), we demonstrate that RAS-dependent suppression of PAR3 (encoded by PARD3) accelerates epithelial disorganization during early tumorigenesis. Diminished PAR3 led to loss of E-cadherin-mediated cell adhesion, which in turn contributed to misoriented cell division. Pharmacological inhibition of the MAPK pathway downstream of RAS activation reversed the defects in PAR3 expression, E-cadherin-mediated cell adhesion and mitotic spindle orientation. Thus, temporal analysis of human neoplasia provides a powerful approach to study cellular and molecular transformations during early oncogenesis, which allowed identification of PAR3 as a critical regulator of tissue architecture during initial human SCC development.

scholarly journals The interaction between the tumour suppressor Dlg1 and the MAGUK protein CASK is required for oriented cell division in mammalian epithelia

2018 ◽  
Author(s):  
Andrew P. Porter ◽  
Gavin White ◽  
Natalie A. Mack ◽  
Angeliki Malliri
Keyword(s):  
Cell Division ◽  
Epithelial Cells ◽  
3D Culture ◽  
Direct Interaction ◽  
Tumour Suppressor ◽  
Spindle Orientation ◽  
Oriented Cell Division ◽  
Cell Divisions ◽  
Microtubule Stability ◽  
Maguk Protein

AbstractOriented cell divisions are important for the formation of normal epithelial structures. Dlg1, a tumour suppressor, is required for oriented cell division in Drosophila epithelia and chick neuroepithelia, but how Dlg1 is localised to the membrane and its importance in mammalian epithelia are unknown. Here we show that Dlg1 is required in non-transformed mammalian epithelial cells for oriented cell divisions, and for normal lumen formation in 3D culture. We demonstrate that CASK, a membrane-associated scaffold, is the factor responsible for Dlg1 membrane localisation during spindle orientation, and thereby identify a new cellular function for CASK. We show that depletion of CASK leads to misoriented divisions in 3D, and to the formation of multilumen structures in cultured kidney and breast epithelial cells. Blocking the direct interaction between CASK and Dlg1 with an interfering peptide disrupts spindle orientation and causes multilumen formation. We further show that the Dlg1-CASK interaction is important for the membrane localisation of the canonical LGN-NuMA complex, required for attachment of the mitotic spindle to the membrane and its correct positioning, as well as for astral microtubule stability. Together these results establish the importance of the CASK-Dlg1 interaction in oriented cell division and epithelial integrity.

Fine Structure of Cytochalasin Induced Polyploid Cells

1968 ◽  
Vol 26 ◽  
pp. 122-123
Author(s):  
Awtar Krishan ◽  
Nestor Bohonos
Keyword(s):  
Fine Structure ◽  
Cytochalasin B ◽  
Present Report ◽  
Cell Monolayer ◽  
Polyploid Cell ◽  
Specific Cell ◽  
Nuclear Surface ◽  
Unsuccessful Attempt ◽  
Daughter Cells ◽  
Polyploid Cells

Cytochalasin B, a mould metabolite from Helminthosporium dermatioideum has been shown to interfere with specific cell activities such as cytoplasmic cleavage and cell movement. Cells undergoing nuclear division in the presence of cytochalasin B are unable to complete the separation of the resulting daughter cells. In time-lapse studies, the daughter cells coalesce after an initial unsuccessful attempt at separation and form large multinucleate polyploid cells. The present report describes the fine structure of the large polyploid cells induced in Earle's L-cell monolayer cultures by exposure to cytochalasin B (lγ/ml) for 92 hours.In the present material we have seen as many as 7 nuclei in these polyploid cells. Treatment with cytochalasin B for longer periods of time (6 to 7 days, with one medium change on the 3rd day) did not increase the number of nuclei beyond the 7 nuclei stage. Figure 1 shows a large polyploid cell with four nuclei. These nuclei are indistinguishable in their fine structure from those of the cells from control cultures but often show unusually large numbers of cytoplasmic invaginations and extensions of the nuclear surface (Figure 2).

Processing Immunoperoxidase Labeled Cell Monolayers and Cryostat Sesctions For Electron Microscopy

1985 ◽  
Vol 43 ◽  
pp. 714-715
Author(s):  
K. Chien ◽  
R. Van de Velde ◽  
I.P. Shintaku ◽  
A.F. Sassoon
Keyword(s):  
Cell Monolayer ◽  
Cell Types ◽  
Surface Antigens ◽  
Immunoperoxidase Method ◽  
Reaction Step ◽  
Hot Plate ◽  
Air Drying ◽  
New Approach ◽  
Cell Monolayers ◽  
Glass Slides

Immunoelectron microscopy of neoplastic lymphoma cells is valuable for precise localization of surface antigens and identification of cell types. We have developed a new approach in which the immunohistochemical staining can be evaluated prior to embedding for EM and desired area subsequently selected for ultrathin sectioning.A freshly prepared lymphoma cell suspension is spun onto polylysine hydrobromide- coated glass slides by cytocentrifugation and immediately fixed without air drying in polylysine paraformaldehyde (PLP) fixative. After rinsing in PBS, slides are stained by a 3-step immunoperoxidase method. Cell monolayer is then fixed in buffered 3% glutaraldehyde prior to DAB reaction. After the DAB reaction step, wet monolayers can be examined under LM for presence of brown reaction product and selected monolayers then processed by routine methods for EM and embedded with the Chien Re-embedding Mold. After the polymerization, the epoxy blocks are easily separated from the glass slides by heatingon a 100°C hot plate for 20 seconds.

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