Ouch! Single Cell Wound Repair in Drosophila Embryos

SciVee ◽  
2011 ◽  
Author(s):  
M.t. Abreu-blanco ◽  
J.m. Verboon ◽  
S.m. Parkhurst
Keyword(s):  
Single Cell ◽  
Wound Repair ◽  
Drosophila Embryos

scholarly journals Microfluidic guillotine for single-cell wound repair studies

2017 ◽  
Vol 114 (28) ◽  
pp. 7283-7288 ◽  
Author(s):  
Lucas R. Blauch ◽  
Ya Gai ◽  
Jian Wei Khor ◽  
Pranidhi Sood ◽  
Wallace F. Marshall ◽  
...  
Keyword(s):  
Single Cell ◽  
High Throughput ◽  
Wound Repair ◽  
Time Course ◽  
Single Cells ◽  
Repair Process ◽  
Gene Knockdown ◽  
Viscous Stress ◽  
Repair Capacity ◽  
Stentor Coeruleus

Wound repair is a key feature distinguishing living from nonliving matter. Single cells are increasingly recognized to be capable of healing wounds. The lack of reproducible, high-throughput wounding methods has hindered single-cell wound repair studies. This work describes a microfluidic guillotine for bisecting single Stentor coeruleus cells in a continuous-flow manner. Stentor is used as a model due to its robust repair capacity and the ability to perform gene knockdown in a high-throughput manner. Local cutting dynamics reveals two regimes under which cells are bisected, one at low viscous stress where cells are cut with small membrane ruptures and high viability and one at high viscous stress where cells are cut with extended membrane ruptures and decreased viability. A cutting throughput up to 64 cells per minute—more than 200 times faster than current methods—is achieved. The method allows the generation of more than 100 cells in a synchronized stage of their repair process. This capacity, combined with high-throughput gene knockdown in Stentor, enables time-course mechanistic studies impossible with current wounding methods.

scholarly journals Cell wound repair in Drosophila occurs through three distinct phases of membrane and cytoskeletal remodeling

2011 ◽  
Vol 193 (3) ◽  
pp. 455-464 ◽  
Author(s):  
Maria Teresa Abreu-Blanco ◽  
Jeffrey M. Verboon ◽  
Susan M. Parkhurst
Keyword(s):  
Cell Death ◽  
Plasma Membrane ◽  
Single Cell ◽  
Wound Repair ◽  
Single Cells ◽  
Repair Process ◽  
Wound Edge ◽  
Cytoskeletal Remodeling ◽  
Tissue Integrity ◽  
E Cadherin

When single cells or tissues are injured, the wound must be repaired quickly in order to prevent cell death, loss of tissue integrity, and invasion by microorganisms. We describe Drosophila as a genetically tractable model to dissect the mechanisms of single-cell wound repair. By analyzing the expression and the effects of perturbations of actin, myosin, microtubules, E-cadherin, and the plasma membrane, we define three distinct phases in the repair process—expansion, contraction, and closure—and identify specific components required during each phase. Specifically, plasma membrane mobilization and assembly of a contractile actomyosin ring are required for this process. In addition, E-cadherin accumulates at the wound edge, and wound expansion is excessive in E-cadherin mutants, suggesting a role for E-cadherin in anchoring the actomyosin ring to the plasma membrane. Our results show that single-cell wound repair requires specific spatial and temporal cytoskeleton responses with distinct components and mechanisms required at different stages of the process.

A single cell approach to problems of cell lineage and commitment during embryogenesis of Drosophila melanogaster

Development ◽  
1987 ◽  
Vol 100 (1) ◽  
pp. 1-12 ◽  
Author(s):  
G.M. Technau
Keyword(s):  
Drosophila Melanogaster ◽  
Single Cell ◽  
Cell Lineage ◽  
Cellular Level ◽  
Central Importance ◽  
Cell Level ◽  
Combined Application ◽  
A Cell ◽  
Pole Cells ◽  
Drosophila Embryos

The mechanisms leading to the commitment of a cell to a particular fate or to restrictions in its developmental potencies represent a problem of central importance in developmental biology. Both at the genetic and at the molecular level, studies addressing this topic using the fruitfly Drosophila melanogaster have advanced substantially, whereas, at the cellular level, experimental techniques have been most successfully applied to organisms composed of relatively large and accessible cells. The combined application of the different approaches to one system should improve our understanding of the process of commitment as a whole. Recently, a method has been devised to study cell lineage in Drosophila embryos at the single cell level. This method has been used to analyse the lineages, as well as the state of commitment of single cell progenitors from various ectodermal, mesodermal and endodermal anlagen and of the pole cells. The results obtained from a clonal analysis of wild-type larval structures are discussed in this review.

scholarly journals Actin and myosin dynamics are independent during Drosophila embryonic wound repair

2019 ◽  
Vol 30 (23) ◽  
pp. 2901-2912
Author(s):  
Anna B. Kobb ◽  
Katheryn E. Rothenberg ◽  
Rodrigo Fernandez-Gonzalez
Keyword(s):  
Wound Repair ◽  
Wound Closure ◽  
Molecular Motor ◽  
Wound Edge ◽  
Cell Movements ◽  
Nonmuscle Myosin Ii ◽  
Wound Margin ◽  
Wound Healing Response ◽  
Disease Cell ◽  
Drosophila Embryos

Collective cell movements play a central role in embryonic development, tissue repair, and metastatic disease. Cell movements are often coordinated by supracellular networks formed by the cytoskeletal protein actin and the molecular motor nonmuscle myosin II. During wound closure in the embryonic epidermis, the cells around the wound migrate collectively into the damaged region. In Drosophila embryos, mechanical tension stabilizes myosin at the wound edge, facilitating the assembly of a supracellular myosin cable around the wound that coordinates cell migration. Here, we show that actin is also stabilized at the wound edge. However, loss of tension or myosin activity does not affect the dynamics of actin at the wound margin. Conversely, pharmacological stabilization of actin does not affect myosin levels or dynamics around the wound. Together, our data suggest that actin and myosin are independently regulated during embryonic wound closure, thus conferring robustness to the embryonic wound healing response.

scholarly journals Actin Cytoskeletal Dynamics in Single-Cell Wound Repair

2021 ◽  
Vol 22 (19) ◽  
pp. 10886
Author(s):  
Malene Laage Ebstrup ◽  
Catarina Dias ◽  
Anne Sofie Busk Heitmann ◽  
Stine Lauritzen Sønder ◽  
Jesper Nylandsted
Keyword(s):  
Plasma Membrane ◽  
Actin Cytoskeleton ◽  
Single Cell ◽  
Wound Repair ◽  
Membrane Integrity ◽  
Limited Attention ◽  
Membrane Repair ◽  
Cortical Actin ◽  
Comprehensive Overview ◽  
Repair Mechanisms

The plasma membrane protects the eukaryotic cell from its surroundings and is essential for cell viability; thus, it is crucial that membrane disruptions are repaired quickly to prevent immediate dyshomeostasis and cell death. Accordingly, cells have developed efficient repair mechanisms to rapidly reseal ruptures and reestablish membrane integrity. The cortical actin cytoskeleton plays an instrumental role in both plasma membrane resealing and restructuring in response to damage. Actin directly aids membrane repair or indirectly assists auxiliary repair mechanisms. Studies investigating single-cell wound repair have often focused on the recruitment and activation of specialized repair machinery, despite the undeniable need for rapid and dynamic cortical actin modulation; thus, the role of the cortical actin cytoskeleton during wound repair has received limited attention. This review aims to provide a comprehensive overview of membrane repair mechanisms directly or indirectly involving cortical actin cytoskeletal remodeling.

scholarly journals Chemical operations on a living single cell by open microfluidics for wound repair studies and organelle transport analysis

2019 ◽  
Vol 10 (7) ◽  
pp. 2081-2087 ◽  
Author(s):  
Sifeng Mao ◽  
Qiang Zhang ◽  
Wu Liu ◽  
Qiushi Huang ◽  
Mashooq Khan ◽  
...  
Keyword(s):  
Laminar Flow ◽  
Single Cell ◽  
Wound Repair ◽  
Organelle Transport ◽  
Transport Analysis ◽  
Living Single

We report a laminar flow based approach that is capable of precisely cutting off or treating a portion of a single cell from its remaining portion in its original adherent state.

scholarly journals A mathematical model of GTPase pattern formation during single-cell wound repair

2016 ◽  
Vol 6 (5) ◽  
pp. 20160032 ◽  
Author(s):  
William R. Holmes ◽  
Adriana E. Golding ◽  
William M. Bement ◽  
Leah Edelstein-Keshet
Keyword(s):  
Single Cell ◽  
Wound Repair ◽  
Critical Role ◽  
Experimental Manipulation ◽  
Cell Polarization ◽  
Wound Response ◽  
Dominant Negative ◽  
Nucleotide Exchange ◽  
Model Approximation ◽  
Positive Feedbacks

Rho GTPases are regulatory proteins whose patterns on the surface of a cell affect cell polarization, cell motility and repair of single-cell wounds. The stereotypical patterns formed by two such proteins, Rho and Cdc42, around laser-injured frog oocytes permit experimental analysis of GTPase activation, inactivation, segregation and crosstalk. Here, we review the development and analysis of a spatial model of GTPase dynamics that describe the formation of concentric zones of Rho and Cdc42 activity around wounds, and describe how this model has provided insights into the roles of the GTPase effector molecules protein kinase C (PKCβ and PKCη) and guanosine nucleotide dissociation inhibitor (GDI) in the wound response. We further demonstrate how the use of a ‘sharp switch’ model approximation in combination with bifurcation analysis can aid mapping the model behaviour in parameter space (approximate results confirmed with numerical simulation methods). Using these methods in combination with experimental manipulation of PKC activity (PKC overexpression (OE) and dominant negative conditions), we have shown that: (i) PKCβ most probably acts by enhancing existing positive feedbacks (from Rho to itself via the guanosine nucleotide exchange factor domain of Abr, and from Cdc42 to itself), (ii) PKCη most probably increases basal rates of inactivation (or possibly decreases basal rates of activation) of Rho and Cdc42, and (iii) the graded distribution of PKCη and its effect on initial Rho activity accounts for inversion of zones in a fraction (20%) of PKCη OE cells. Finally, we speculate that GDIs (which sequester GTPases) may have a critical role in defining the spatial domain, where the wound response may occur. This paper provides a more thorough exposition of the methods of analysis used in the investigation, whereas previous work on this topic was addressed to biologists and abbreviated such discussion.

scholarly journals Planar Cell Polarity and E-Cadherin in Tissue-Scale Shape Changes in Drosophila Embryos

2020 ◽  
Vol 8 ◽  
Author(s):  
Deqing Kong ◽  
Jörg Großhans
Keyword(s):  
Cell Polarity ◽  
Wound Repair ◽  
Large Scale ◽  
Planar Cell Polarity ◽  
Cell Communication ◽  
Cell Dynamics ◽  
Axis Elongation ◽  
Shape Changes ◽  
Drosophila Embryos ◽  
E Cadherin

Planar cell polarity and anisotropic cell behavior play critical roles in large-scale epithelial morphogenesis, homeostasis, wound repair, and regeneration. Cell–Cell communication and mechano-transduction in the second to minute scale mediated by E-cadherin complexes play a central role in the coordination and self-organization of cellular activities, such as junction dynamics, cell shape changes, and cell rearrangement. Here we review the current understanding in the interplay of cell polarity and cell dynamics during body axis elongation and dorsal closure in Drosophila embryos with a focus on E-cadherin dynamics in linking cell and tissue polarization and tissue-scale shape changes.

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