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Roubinet, C.

Publications and source records attributed to Roubinet, C..

5 recordsLinked to original sources

Cell size overrides ganglion mother cell fate specification in Drosophila

The size of cells varies enormously in biology and how cell size and function are linked are fundamental questions. Asymmetrically dividing Drosophila neural stem cells, also called neuroblasts, produce a large neuroblast and a smaller ganglion mother cell (GMC) at each division. However, why GMCs need to be smaller than neuroblasts has largely remained unexplained. Here, we developed acute and reversible methods to alter daughter-cell size during a single neuroblast division while preserving cortical polarity and asymmetric inheritance of fate determinants. We identify atypical protein kinase C (aPKC) as a regulator of size asymmetry and show that transient and partial co-inhibition of aPKC and Rho kinase (Drok in Drosophila) generates daughters spanning a size range, including size-wise near-symmetric sibling cells while polarity is preserved. We find that above a certain size threshold daughter cells retain neuroblast-like properties despite inheriting GMC determinants: they divide repeatedly, exhibit neuroblast-like cell-cycle timing, and undergo size-wise asymmetric division. Smaller daughters below this threshold instead follow a GMC-like trajectory. Thus, in Drosophila type I neuroblasts, daughter-cell size can act as a regulator of lineage behaviour, overriding inherited fate cues in sufficiently large daughter cells.

developmental biology↗

The transmembrane protein Syndecan regulates stem cell nuclear properties and cell maintenance.

Tissue maintenance is underpinned by resident stem cells whose activity is modulated by microenvironmental cues. Using Drosophila as a simple model to identify regulators of stem cell behaviour and survival in vivo, we have identified novel connections between the conserved transmembrane proteoglycan Syndecan, nuclear properties and stem cell function. In the Drosophila midgut, Syndecan depletion in intestinal stem cells results in their loss from the tissue, impairing tissue renewal. At the cellular level, Syndecan depletion alters cell and nuclear shape, and causes nuclear lamina invaginations and DNA damage. In a second tissue, the developing Drosophila brain, live imaging revealed that Syndecan depletion in neural stem cells results in nuclear envelope remodelling defects which arise upon cell division. Our findings reveal a new role for Syndecan in the maintenance of nuclear properties in diverse stem cell types. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=198 SRC="FIGDIR/small/580237v1_ufig1.gif" ALT="Figure 1"> View larger version (60K): org.highwire.dtl.DTLVardef@14775a4org.highwire.dtl.DTLVardef@7c66beorg.highwire.dtl.DTLVardef@13d5dd7org.highwire.dtl.DTLVardef@1dbd0f0_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

SCAR and the Arp2/3 complex polarise the actomyosin cortex and plasma membrane organization in asymmetrically dividing neuroblasts

While the Formin-nucleated actomyosin cortex has been shown to drive the changes in cell shape that accompany cell division in both symmetric and asymmetric cell divisions, it is not clear whether or not Arp2/3-nucleated branched actin filament networks also play a role. In order to look for mitotic roles of the Arp2/3 complex, here we use Drosophila neural stem cells as a model system. These cells are unusual in that they divide asymmetrically to produce a large and small daughter cell with different fates. Our analysis identifies a pool of Arp2/3-dependent actin-based membrane protrusions that form at the apical cortex of these cells as they enter mitosis. Strikingly, at metaphase, these protrusions co-localise with components of the SCAR complex. By perturbing Arp2/3 complex activity we show that this apical pool of actin likely functions to limit the accumulation of apical Myosin in metaphase. Following the onset of anaphase, the loss of these SCAR and Arp2/3 dependent structures then leads to a delay in the clearance of apical Myosin and to cortical instability at cytokinesis. These data point to a role for a polarised branched actin filament network in fine tuning the apical actomyosin cortex to enable the precise control of cell shape during asymmetric cell division.

cell biology↗

Asymmetric nuclear division of neural stem cells contributes to the formation of sibling nuclei with different identities

Cellular diversity in multicellular organisms is often generated via asymmetric divisions. In the fly, for example, neural stem cells divide asymmetrically to generate a large self-renewing stem cell and a smaller sibling that differentiates. Efforts to understand how these different cell fates are generated have focused on the asymmetric segregation of cortically-localised transcription factors at division, which preferentially enter single daughter cell nuclei to change their fate. However, we find that the nuclear compartment in these cells remains intact throughout mitosis and is asymmetrically inherited, giving rise to sibling nuclei that differ profoundly in size, envelope composition and fate markers. These data reveal the importance of considering nuclear remodelling during stem cell divisions, and show how daughter cell fates depend on the coordination of the asymmetric inheritance of cortical fate markers with asymmetric nuclear division.

cell biology↗

Live cell imaging of the hyperthermophilic archaeon Sulfolobus acidocaldarius identifies complementary roles for two ESCRTIII homologues in ensuring a robust and symmetric cell division

Live-cell imaging has revolutionized our understanding of dynamic cellular processes in bacteria and eukaryotes. While similar techniques have recently been applied to the study of halophilic archaea, our ability to explore the cell biology of thermophilic archaea is limited, due to the technical challenges of imaging at high temperatures. Here, we report the construction of the Sulfoscope, a heated chamber that enables live-cell imaging on an inverted fluorescent microscope. Using this system combined with thermostable fluorescent probes, we were able to image Sulfolobus cells as they divide, revealing a tight coupling between changes in DNA compaction, segregation and cytokinesis. By imaging deletion mutants, we observe important differences in the function of the two ESCRTIII proteins recently implicated in cytokinesis. The loss of CdvB1 compromises cell division, causing occasional division failures and fusion of the two daughter cells, whereas the deletion of cdvB2 leads to a profound loss of division symmetry, generating daughter cells that vary widely in size and eventually generating ghost cells. These data indicate that DNA separation and cytokinesis are coordinated in Sulfolobus, as is the case in eukaryotes, and that two contractile ESCRTIII polymers perform distinct roles to ensure that Sulfolobus cells undergo a robust and symmetrical division. Taken together, the Sulfoscope has shown to provide a controlled high temperature environment, in which cell biology of Sulfolobus can be studied in unprecedent details.

microbiology↗