During gastrulation in Drosophila melanogaster, coordinated apical constriction of the cellular surface drives invagination of the mesoderm anlage. Forces generated by the cortical cytoskeletal network have a pivotal role in this cellular shape change. Here, we show that the organisation of cortical actin is essential for stabilisation of the cellular surface against contraction. We found that mutation of genes related to heterotrimeric G protein (HGP) signaling, such as Gβ13F, Gγ1, and ric-8, results in formation of blebs on the ventral cellular surface. The formation of blebs is caused by perturbation of cortical actin and induced by local surface contraction. HGP signaling mediated by two Gα subunits, Concertina and G-iα65A, constitutively regulates actin organisation. We propose that the organisation of cortical actin by HGP is required to reinforce the cortex so that the cells can endure hydrostatic stress during tissue folding.

A central problem in biomedical imaging is the automated segmentation of images for further quantitative analysis. Recently, fully convolutional neural networks, such as the U-Net, were applied successfully in a variety of segmentation tasks. A downside of this approach is the requirement for a large amount of well-prepared training samples, consisting of image -ground truth mask pairs. Since training data must be created by hand for each experiment, this task can be very costly and time-consuming. Here, we present a segmentation method based on cycle consistent generative adversarial networks, which can be trained even in absence of prepared image -mask pairs. We show that it successfully performs image segmentation tasks on samples with substantial defects and even generalizes well to different tissue types.

AbstractsThe spatial and temporal dynamics of cell contractility plays a key role in tissue morphogenesis, wound healing and cancer invasion. Here we report a simple, single cell resolution, optochemical method to induce minute-scale cell contractions in vivo during morphogenesis. We employed the photolabile Ca 2+ chelator o-nitrophenyl EGTA to induce bursts of intracellular free Ca 2+ by laser photolysis. Ca 2+ bursts appear within seconds and are restricted to individual target cells. Cell contraction reliably followed within a minute, to about half of the cross-sectional area. Increased Ca 2+ levels and contraction were reversible and the target cells further participated in tissue morphogenesis. Depending on Rho kinase (Rok) activity but not RhoGEF2, cell contractions are paralleled with non-muscle myosin-II accumulation in the apicomedial cortex, indicating that Ca 2+ bursts trigger non-muscle myosin II activation. Our approach can be easily adapted to many experimental systems and species, as no specific genetic elements are required and a widely used reagent is employed.

Cell cycle remodeling from fast nuclear cycles to a generic cell cycle mode is a major feature of the mid-blastula transition (MBT) in Drosophila. Remodeling occurs when Twine/Cdc25 falls below a critical threshold. Timing is based on Twine destabilization induced by zygotic transcription. It is conceivable that appropriate starting levels are also important for timely reaching the threshold. Mechanisms for controlling Twine levels at the onset of MBT are unknown. Here we identify a function of the protein phosphatase V in this mechanism. Twine was increased in PpV mutants, whereas the decay rate was comparable to wildtype. PpV mutants frequently underwent an extra nuclear division. We detected PpV-dependent phosphosites in Twine. Phosphosite mutants contain higher Twine levels and frequently underwent an extra nuclear division, comparable to PpV mutants. Our data support a model that the cell cycle remodeling is controlled by induced destabilization and PpV-d ependent control of Twine levels.

Summary statement: Expression of Lamin Dm0 or Kugelkern curb stem cell proliferation by suppression of Jak/Stat signalling. AbstractThe nuclear lamina is involved in numerous cellular functions, such as gene expression, nuclear organization, nuclear stability, and cell proliferation. The mechanism underlying the involvement of lamina is often not clear, especially in physiological contexts. Here we investigate how the farnesylated lamina proteins Lamin Dm0 and Kugelkern are linked to proliferation control of intestinal stem cells (ISCs) in adult Drosophila flies by loss-of-function and gain-of-function experiments.We found that ISCs mutant for Lamin Dm0 or Kugelkern proliferate, whereas overexpression of Lamin Dm0 or Kugelkern strongly suppressed proliferation. The anti-proliferative activity is, at least in part, due to suppression of Jak/Stat but not Delta/Notch signalling. Lamin Dm0 expression suppresses Jak/Stat signalling by normalization of about 50% of the Stat target genes in ISCs.All rights reserved. No reuse allowed without permission.(which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint . http://dx.doi.org/10.1101/177998 doi: bioRxiv preprint first posted online Aug. 18, 2017; Author summaryThe nuclear lamina is a protein meshwork that lies beneath the inner side of the nuclear membrane and interacts with nuclear pores, chromatin and the cytoskeleton.Changes in proteins of the nuclear lamina cause a wide range of diseases which are often not well understood. It is hypothesized that impairment of stem cell function, as a result of lamina changes, might play a key role in some of those diseases. Here we use the well characterized Drosophila midgut as a system to investigate the role of lamina proteins Lamin Dm0 and Kugelkern on stem cell proliferation.