Cellular Rearrangements Research Articles

Nuclei as mechanical bumpers during epithelial remodeling.

The morphogenesis of developing tissues relies on extensive cellular rearrangements in shape, position, and identity. A key process in reshaping tissues is cell intercalation-driven elongation, where epithelial cells align and intercalate along a common axis. Typically, analyses focus on how peripheral cortical forces influence cell shape changes. Less attention is given to how inhomogeneities in internal structures, particularly the nucleus, impact cell shaping. Here, we examine how pulsed contractile and extension dynamics interact with the nucleus in elongating Drosophila embryos. Our data show that tightly packed nuclei in apical layers hinder tissue remodeling/oscillatory behaviors. We identify two mechanisms for resolving internuclear tensions: nuclear deformation and dispersion. Embryos with non-deformable nuclei use nuclear dispersion to maintain near-normal extensile rates, while those with non-dispersible nuclei due to microtubule inhibition exhibit disruptions in contractile behaviors. Disrupting both mechanisms leads to severe tissue extension defects and cell extrusion. These findings highlight the critical role of nuclear shape and positioning in topological remodeling of epithelia.

Symmetry breaking and fate divergence during lateral inhibition in Drosophila.

Lateral inhibition mediates alternative cell fate decision and produces regular cell fate patterns with fate symmetry breaking (SB) relying on the amplification of small stochastic differences in Notch activity via an intercellular negative-feedback loop. Here, we used quantitative live imaging of endogenous Scute (Sc), a proneural factor, and of a Notch activity reporter to study the emergence of sensory organ precursor cells in the pupal abdomen of Drosophila. SB was observed at low Sc levels and was not preceded by a phase of intermediate Sc expression and Notch activity. Thus, mutual inhibition may only be transient in this context. In support of the intercellular feedback loop model, cell-to-cell variations in Sc levels promoted fate divergence. The size of the apical area of competing cells did not detectably bias this fate choice. Surprisingly, cells that were in direct contact at the time of SB could adopt the sensory organ precursor cell fate, albeit at low frequency (10%). These lateral inhibition defects were corrected by cellular rearrangements, not cell fate change, highlighting the role of cell-cell intercalation in pattern refinement.

The Value of Chemical Shift Magnetic Resonance Imaging in Characterization of Bone Tumors

Abstract Background The primary bone tumor incidence rate comprises at least two distinct peaks of an individual's life in the second decade and late adulthood. The dynamic of bone tissue- dependent on the tissue’s physiological and pathological condition which makes it susceptible to cellular re-arrangement, tumor cell formation, and tumor cell implantation. Aim of the Work to investigate the potential application of opposed phase imaging (OPI) or chemical shift imaging (CSI) in characterization and differentiation of various bone tumors to set a cut-off point using chemical shift imaging signal intensity ratio to differentiate benign from malignant bone tumors. Patients and Methods This study is a cross-sectional study was conducted on 35 patients who are clinically suspected to bone tumor, the period from July 2022 to July 2023 in Radio- diagnosis Department of Aim Shams University and Police Academy Hospital was undergoing MRI imaging for the identification of bone tumors. Results The current study findings compared the gender and age distributions among patients with benign and malignant bone lesions. Among the benign group (N = 21), 57.1% were male and 42.9% were female, while in the malignant group (N = 14), 71.4% were male and 28.6% were female. A chi-square test was conducted for gender, yielding a test statistic of 0.734 and a p-value of 0.392, indicating no statistically significant difference in gender distribution between benign and malignant lesions (p > 0.05). Regarding age, the mean age in the benign group was 26.76 years with a standard deviation of 14.20 years, whereas in the malignant group, it was 42.29 years with a standard deviation of 23.6 years. The age difference between the two groups was statistically significant, as determined by an independent samples t-test (T = 2.209, p = 0.039), suggesting that patients with malignant bone lesions tended to be older compared to those with benign lesions. The current study findings compared lesion site distribution between patients with benign and malignant bone lesions. In the benign group (N = 21), the femur was the most common site (28.6%), followed by the tibia (23.8%) and radius (9.5%). No benign lesions were observed in certain sites. In the malignant group (N = 14), the vertebrae were most common (21.4%), followed by the femur and tibia (each 28.6%). A chi-square test showed no significant difference in lesion site distribution between the two groups (p = 0.234). Conclusion the comprehensive analysis of various parameters derived from chemical shift imaging (CSI) in the characterization of bone lesions reveals valuable insights into their diagnostic utility. The study examined demographic characteristics, lesion sites, sizes, and CSI parameters in a cohort of patients with benign and malignant bone lesions. Notably, while gender distribution and lesion sites did not significantly differ between benign and malignant groups, age and CSI parameters demonstrated notable distinctions. Malignant lesions tended to occur in older individuals and exhibited higher mean out-phase signal intensities compared to benign lesions. Importantly, the signal intensity ratio (SIR) emerged as a highly sensitive and specific parameter for distinguishing between benign and malignant lesions, with a perfect diagnostic performance at the specified cutoff value. These findings underscore the importance of incorporating CSI, particularly SIR, into routine imaging protocols for accurate characterization and differentiation of bone lesions, thereby facilitating timely diagnosis and appropriate clinical management.

Three-dimensional stem cell models of mammalian gastrulation.

Gastrulation is a key milestone in the development of an organism. It is a period of cell proliferation and coordinated cellular rearrangement, that creates an outline of the body plan. Our current understanding of mammalian gastrulation has been improved by embryo culture, but there are still many open questions that are difficult to address because of the intrauterine development of the embryos and the low number of specimens. In the case of humans, there are additional difficulties associated with technical and ethical challenges. Over the last few years, pluripotent stem cell models are being developed that have the potential to become useful tools to understand the mammalian gastrulation. Here we review these models with a special emphasis on gastruloids and provide a survey of the methods to produce them robustly, their uses, relationship to embryos, and their prospects as well as their limitations.

Role of tissue fluidization and topological defects in epithelial tubulogenesis

Cellular rearrangements, as primary sources of tissue fluidization, facilitate topological transitions during tissue morphogenesis. We study the role of intrinsic cell properties such as cell polarity and cell-cell adhesion in shaping epithelial tissues using a minimal model of interacting polarized cells. The presence of a vortex in the cell polarization poses the topological constraint that induces an inwards migration with the formation of a conical shape. Local rearrangements at the tip of the cone lead to the onset of tube formation. Switching between collective migration and structural rearrangements is key for balancing the contrasting tendencies, such as the tissue rigidity needed to preserve shape and the tissue fluidity allowing for topological transitions during tissue morphogenesis. Published by the American Physical Society 2024

Myosin II mediates Shh signals to shape dental epithelia via control of cell adhesion and movement.

The development of ectodermal organs begins with the formation of a stratified epithelial placode that progressively invaginates into the underlying mesenchyme as the organ takes its shape. Signaling by secreted molecules is critical for epithelial morphogenesis, but how that information leads to cell rearrangement and tissue shape changes remains an open question. Using the mouse dentition as a model, we first establish that non-muscle myosin II is essential for dental epithelial invagination and show that it functions by promoting cell-cell adhesion and persistent convergent cell movements in the suprabasal layer. Shh signaling controls these processes by inducing myosin II activation via AKT. Pharmacological induction of AKT and myosin II can also rescue defects caused by the inhibition of Shh. Together, our results support a model in which the Shh signal is transmitted through myosin II to power effective cellular rearrangement for proper dental epithelial invagination.

Acinar-ductal cell rearrangement drives branching morphogenesis of the murine pancreas in an IGF/PI3K-dependent manner

During organ formation, progenitor cells need to acquire different cell identities and organize themselves into distinct structural units. How these processes are coordinated and how tissue architecture(s) is preserved despite the dramatic cell rearrangements occurring in developing organs remain unclear. Here, we identified cellular rearrangements between acinar and ductal progenitors as a mechanism to drive branching morphogenesis in the pancreas while preserving the integrity of the acinar-ductal functional unit. Using exvivo and invivo mouse models, we found that pancreatic ductal cells form clefts by protruding and pulling on the acinar basement membrane, which leads to acini splitting. Newly formed acini remain connected to the bifurcated branches generated by ductal cell rearrangement. Insulin growth factor (IGF)/phosphatidylinositol 3-kinase (PI3K) pathway finely regulates this process by controlling pancreatic ductal tissue fluidity, with a simultaneous impact on branching and cell fate acquisition. Together, our results explain how acinar structure multiplication and branch bifurcation are synchronized during pancreas organogenesis.

Inhibition of the SREBP pathway prevents SARS-CoV-2 replication and inflammasome activation.

SARS-CoV-2 induces major cellular lipid rearrangements, exploiting the host's metabolic pathways to replicate. Sterol regulatory element binding proteins (SREBPs) are a family of transcription factors that control lipid metabolism. SREBP1 is associated with the regulation of fatty acids, whereas SREBP2 controls cholesterol metabolism, and both isoforms are associated with lipid droplet (LD) biogenesis. Here, we evaluated the effect of SREBP in a SARS-CoV-2-infected lung epithelial cell line (Calu-3). We showed that SARS-CoV-2 infection induced the activation of SREBP1 and SREBP2 and LD accumulation. Genetic knockdown of both SREBPs and pharmacological inhibition with the dual SREBP activation inhibitor fatostatin promote the inhibition of SARS-CoV-2 replication, cell death, and LD formation in Calu-3 cells. In addition, we demonstrated that SARS-CoV-2 induced inflammasome-dependent cell death by pyroptosis and release of IL-1β and IL-18, with activation of caspase-1, cleavage of gasdermin D1, was also reduced by SREBP inhibition. Collectively, our findings help to elucidate that SREBPs are crucial host factors required for viral replication and pathogenesis. These results indicate that SREBP is a host target for the development of antiviral strategies.

Biomimetic Fibers Derived from an Equidistant Micropillar Platform Dictate Osteocyte Fate via Mechanoreception.

It is a big challenge to design a biomimetic physical microenvironment with greater similarity to in vivo tissue to observe real cell behaviors. We established a novel cell culture platform based on patterned equidistant micropillars with stiff and soft stiffnesses to mimic the changes that happened in the transition from normal to osteoporotic disease. We first demonstrated that the soft micropillar substrate decreased osteocyte synaptogenesis through synaptogyrin 1 and that this decrease was accompanied by impairment of cell mechanoperception and a decrease in cellular cytoskeletal rearrangement. We then found that the soft equidistant micropillar substrate reduced the osteocyte synaptogenesis mainly via the inactivation of Erk/MAPK signaling. We finally found that soft micropillar substrate-mediated synaptogenesis impacted the cell-to-cell communication and matrix mineralization of osteocytes. Taken together, this study provides evidence of cellular mechanical responses that are much more similar to those of real osteocytes at the bone tissue level.

Robust statistical properties of T1 transitions in a multi-phase field model of cell monolayers

Large-scale tissue deformation which is fundamental to tissue development hinges on local cellular rearrangements, such as T1 transitions. In the realm of the multi-phase field model, we analyse the statistical and dynamical properties of T1 transitions in a confluent monolayer. We identify an energy profile that is robust to changes in several model parameters. It is characterized by an asymmetric profile with a fast increase in energy before the T1 transition and a sudden drop after the T1 transition, followed by a slow relaxation. The latter being a signature of the fluidity of the cell monolayer. We show that T1 transitions are sources of localised large deformation of the cells undergoing the neighbour exchange, and they induce other T1 transitions in the nearby cells leading to a chaining of events that propagate local cell deformation to large scale tissue flows.

The "sweet" path to cancer: focus on cellular glucose metabolism.

The hypoxia-inducible factor-1α (HIF-1α), a key player in the adaptive regulation of energy metabolism, and the M2 isoform of the glycolytic enzyme pyruvate kinase (PKM2), a critical regulator of glucose consumption, are the main drivers of the metabolic rewiring in cancer cells. The use of glycolysis rather than oxidative phosphorylation, even in the presence of oxygen (i.e., Warburg effect or aerobic glycolysis), is a major metabolic hallmark of cancer. Aerobic glycolysis is also important for the immune system, which is involved in both metabolic disorders development and tumorigenesis. More recently, metabolic changes resembling the Warburg effect have been described in diabetes mellitus (DM). Scientists from different disciplines are looking for ways to interfere with these cellular metabolic rearrangements and reverse the pathological processes underlying their disease of interest. As cancer is overtaking cardiovascular disease as the leading cause of excess death in DM, and biological links between DM and cancer are incompletely understood, cellular glucose metabolism may be a promising field to explore in search of connections between cardiometabolic and cancer diseases. In this mini-review, we present the state-of-the-art on the role of the Warburg effect, HIF-1α, and PKM2 in cancer, inflammation, and DM to encourage multidisciplinary research to advance fundamental understanding in biology and pathways implicated in the link between DM and cancer.

Design, Modeling, and Application of Reinforced-Airbag-Based Pneumatic Actuators with High Load and Cellular Rearrangement.

Although various soft pneumatic actuators have been studied, their performance, including load capacity, has not been satisfied yet. Enhancing their actuation capability and using them to develop soft robots with high performance is still an open and challenging issue. In this study, we developed novel pneumatic actuators based on fiber-reinforced airbags as a solution to this problem, of which the maximum pressure reaches more than 100 kPa. Through cellular rearrangement, the developed actuators could bend uni- or bidirectionally, achieving large driving force, large deformation, and high conformability. Hence, they could be used to develop soft manipulators with relatively large payload (up to 10 kg, about 50 times the body self-weight) and soft climbing robots with high mobility. In this article, we first present the design of the airbag-based actuators and then model the airbag to obtain the relationship between the pneumatic pressure, external force, and deformation. Subsequently, we validate the models by comparing the simulated and measured results and test the load capacity of the bending actuators. Afterward, we present the development of a soft pneumatic robot that can rapidly climb horizontal, inclined, and vertical poles with different cross-sectional shapes and even outdoor natural objects, like bamboos, at a speed of 12.6 mm/s generally. In particular, it can dexterously transition between poles at any angle, which, to the best of our knowledge, has not been achieved before.

Understanding the effect of temperature downshift on CHO cell growth, antibody titer and product quality by intracellular metabolite profiling and in vivo monitoring of redox state.

The strategy of temperature downshift has been widely used in the biopharmaceutical industry to improve antibody production and cell-specific production rate (qp ) with Chinese hamster ovary cells (CHO). However, the mechanism of temperature-induced metabolic rearrangement, especially important intracellular metabolic events, remains poorly understood. In this work, in order to explore the mechanisms of temperature-induced cell metabolism, we systematically assessed the differences in cell growth, antibody expression, and antibody quality between high-producing (HP) and low-producing (LP) CHO cell lines under both constant temperature (37°C) and temperature downshift (37°C→33°C) settings during fed-batch culture. Although the results showed that low-temperature culture during the late phase of exponential cell growth significantly reduced the maximum viable cell density (p < 0.05) and induced cell cycle arrest in the G0/G1 phase, this temperature downshift led to a higher cellular viability and increased antibody titer by 48% and 28% in HP and LP CHO cell cultures, respectively (p < 0.001), and favored antibody quality reflected in reduced charge heterogeneity and molecular size heterogeneity. Combined extra- and intra-cellular metabolomics analyses revealed that temperature downshift significantly downregulated intracellular glycolytic and lipid metabolic pathways while upregulated tricarboxylic acid (TCA)cycle, and particularly featured upregulated glutathione metabolic pathways. Interestingly, all these metabolic pathways were closely associated with the maintenance of intracellular redox state and oxidative stress-alleviating strategies. To experimentally address this, we developed two high-performance fluorescent biosensors, denoted SoNar and iNap1, for real-time monitoring of intracellular nicotinamide adenine dinucleotide/nicotinamide adenine dinucleotide + hydrogen (NAD+ /NADH) ratio and nicotinamide adenine dinucleotide phosphate (NADPH) amount, respectively. Consistent with such metabolic rearrangements, the results showed that temperature downshift decreased the intracellular NAD+ /NADH ratio, which might be ascribed to the re-consumption of lactate, and increased the intracellular NADPH amount (p < 0.01) to scavenge intracellular reactive oxygen species (ROS) induced by the increased metabolic requirements for high-level expression of antibody. Collectively, this study provides a metabolic map of cellular metabolic rearrangement induced by temperature downshift and demonstrates the feasibility of real-time fluorescent biosensors for biological processes, thus potentially providing a new strategy for dynamic optimization of antibody production processes.

Regulation of aortic morphogenesis and VE-cadherin dynamics by VEGF.

In amniote vertebrates, the definitive dorsal aorta is formed by the fusion of two primordial aortic endothelial tubes. Formation of the definitive dorsal aorta requires extensive cellular migrations and rearrangements of the primordial tubes in order to generate a single vessel located at the embryonic ventral midline. This study examines the role of VEGF signaling in the generation of the definitive dorsal aorta. Through gain- and loss-of-function studies in vivo in the chick embryo, we document a requirement for VEGF signaling in growth and remodeling of the paired primordia. We find that regions of the aorta are differentially sensitive to levels of VEGF signaling, and present evidence that areas of low blood flow are more sensitive to the loss of VEGF signaling. We also find that VEGF signaling regulates the intracellular distribution between membrane and cytoplasm of the cell-cell adhesion molecule VE-cadherin in aortic endothelial cells in vivo. Together, these finding identify mechanisms that likely contribute to the dynamic behavior of endothelial cells during aorta morphogenesis.

Morphogenesis-inspired two-dimensional electrowetting in droplet networks

Living tissues dynamically reshape their internal cellular structures through carefully regulated cell-to-cell interactions during morphogenesis. These cellular rearrangement events, such as cell sorting and mutual tissue spreading, have been explained using the differential adhesion hypothesis, which describes the sorting of cells through their adhesive interactions with their neighbors. In this manuscript we explore a simplified form of differential adhesion within a bioinspired lipid-stabilized emulsion approximating cellular tissues. The artificial cellular tissues are created as a collection of aqueous droplets adhered together in a network of lipid membranes. Since this abstraction of the tissue does not retain the ability to locally vary the adhesion of the interfaces through biological mechanisms, instead we employ electrowetting with offsets generated by spatial variations in lipid compositions to capture a simple form of bioelectric control over the tissue characteristics. This is accomplished by first conducting experiments on electrowetting in droplet networks, next creating a model for describing electrowetting in collections of adhered droplets, then validating the model against the experimental measurements. This work demonstrates how the distribution of voltage within a droplet network may be tuned through lipid composition then used to shape directional contraction of the adhered structure using two-dimensional electrowetting events. Predictions from this model were used to explore the governing mechanics for complex electrowetting events in networks, including directional contraction and the formation of new interfaces.

Host-Erythrocytic Sphingosine-1-Phosphate Regulates Plasmodium Histone Deacetylase Activity and Exhibits Epigenetic Control over Cell Death and Differentiation.

The evolution of resistance to practically all antimalarial drugs poses a challenge to the current malaria elimination and eradication efforts. Given that the epigenome of Plasmodium falciparum governs several crucial parasite functions, pharmaceutical interventions with transmission-blocking potential that target epigenetic molecular markers and regulatory mechanisms are likely to encounter drug resistance. In the malaria parasite, histone deacetylases (HDACs) are essential epigenetic modulators that regulate cellular transcriptional rearrangements, notably the molecular mechanisms underlying parasite proliferation and differentiation. We establish "lipid sequestration" as a mechanism by which sphingolipids, specifically Sphingosine-1-Phosphate (S1P) (a metabolic product of Sphingosine Kinase 1 [SphK-1]), regulate epigenetic reprogramming in the parasite by interacting with, and modulating, the histone-deacetylation activity of PfHDAC-1, thereby regulating Plasmodium pathogenesis. Furthermore, we demonstrate that altering host S1P levels with PF-543, a potent and selective Sphk-1 inhibitor, dysregulates PfHDAC-1 activity, resulting in a significant increase in the global histone acetylation signals and, consequently, transcriptional modulation of genes associated with gametocytogenesis, virulence, and proliferation. Our findings point to a hitherto unrecognized functional role for host S1P-mediated sphingolipid signaling in modulating PfHDAC-1's enzymatic activity and, as a result, the parasite's dynamic genome-wide transcriptional patterns. The epigenetic regulation of parasite proliferation and sexual differentiation offers a novel approach for developing host-targeted therapeutics to combat malaria resistance to conventional regimens. IMPORTANCE Sphingolipid is an 18-carbon amino-alcohol-containing lipid with a sphingosine backbone, which when phosphorylated by sphingosine kinase 1 (SphK-1), generates sphingosine-1-phosphate (S1P), an essential lipid signaling molecule. Dysregulation of S1P function has been observed in a variety of pathologies, including severe malaria. The malaria parasite Plasmodium acquires a host S1P pool for its growth and survival. Here, we describe the molecular attuning of histone deacetylase-1 (PfHDAC-1), a crucial epigenetic modulator that contributes to the establishment of epigenetic chromatin states and parasite survival, in response to S1P binding. Our findings highlight the host lipid-mediated epigenetic regulation of malaria parasite key genes.

Human adipose mesenchymal stromal cells growing into PCL-nHA electrospun scaffolds undergo hypoxia adaptive ultrastructural changes.

Human Adipose-Derived Mesenchymal Stem/Stromal Cells (hAD-MSCs) have great potential for tissue regeneration. Since transplanted hAD-MSCs are likely to be placed in a hypoxic environment, culturing the cells under hypoxic conditions might improve their post-transplantation survival and regenerative performance. The combination of hAD-MSCs and PCL-nHA nanofibers synergically improves the contribution of both components for osteoblast differentiation. In this work, we hypothesized that this biomaterial constitutes a hypoxic environment for hAD-MSCs. We studied the cellular re-arrangement and the subcellular ultrastructure by Transmission Electron Microscopy (TEM) of hAD-MSCs grown into PCL-nHA nanofibers, and we compared them with the same cells grown in two-dimensional cultures, over tissue culture-treated plastic, or glass coverslips. Among the most evident changes, PCL-nHA grown cells showed enlarged mitochondria, and accumulation of glycogen granules, consistent with a hypoxic environment. We observed a 3.5 upregulation (p=0.0379) of Hypoxia Inducible Factor (HIF)-1A gene expression in PCL-nHA grown cells. This work evidences for the first time intra-cellular changes in three-dimensional compared to two-dimensional cultures, which are adaptive responses of the cells to an environment more closely resembling that of the in vivo niche after transplantation, thus PCL-nHA nanofibers are adequate for hAD-MSCs pre-conditioning.

Proteomic analysis of fetal skin by iTRAQ reveals molecular signals underlying Inner Mongolia Cashmere goat hair follicle initiation

Cashmere goats produce wool from primary hair follicles (PHF)and cashmere from secondary hair follicles(SHF). Numerous genetic factors have been identified to affect HF formation, but the mechanism that controls the distinct morphogenesis of PHFs and SHFs remains unclear. To better describe goat’s hair follicle proteome, we applied iTRAQ technology to study protein expression profiling from nine skin samples of cashmere goats in three fetal periods including 45-day-old embryos (E45), 55-day-old embryos (E55), 65-day-old embryos (E65). A total of 1172 proteins identified are expressed in fetal skin. The numbers of differentially expressed proteins in E45 vs. E55, E45 vs. E65, and E55 vs. E65 were 103, 144, and 118, respectively. Sixty-three proteins were constantly increased through the whole embryonic developmental stages of placode formation. Keratin 1, 4, 5, 10, 13, 15, 17, and 75 belonging to the hair keratin family (KRTs) were identified during keratinocyte proliferation. Some proteins involved in these pathways probably have a role in the HF and might be the mediators of HF initiation in Cashmere goats. Our systematic investigation identified some signaling factors that drive the large-scale cellular rearrangements necessary for HF initiation, which provide an insight into the fundamental mechanisms of cashmere growth.

Master mitotic kinases regulate viral genome delivery during papillomavirus cell entry

Mitosis induces cellular rearrangements like spindle formation, Golgi fragmentation, and nuclear envelope breakdown. Similar to certain retroviruses, nuclear delivery during entry of human papillomavirus (HPV) genomes is facilitated by mitosis, during which minor capsid protein L2 tethers viral DNA to mitotic chromosomes. However, the mechanism of viral genome delivery and tethering to condensed chromosomes is barely understood. It is unclear, which cellular proteins facilitate this process or how this process is regulated. This work identifies crucial phosphorylations on HPV minor capsid protein L2 occurring at mitosis onset. L2’s chromosome binding region (CBR) is sequentially phosphorylated by the master mitotic kinases CDK1 and PLK1. L2 phosphorylation, thus, regulates timely delivery of HPV vDNA to mitotic chromatin during mitosis. In summary, our work demonstrates a crucial role of mitotic kinases for nuclear delivery of viral DNA and provides important insights into the molecular mechanism of pathogen import into the nucleus during mitosis.

Recoil Measurements in Drosophila Embryos: from Mounting to Image Analysis.

Tension and force propagation play a central role in tissue morphogenesis, as they enable sub- and supra-cellular shape changes required for the generation of new structures. Force is often generated by the cytoskeleton, which forms complex meshworks that reach cell-cell or cell-extracellular matrix junctions to induce cellular rearrangements. These mechanical properties can be measured through laser microdissection, which concentrates energy in the tissue of interest, disrupting its cytoskeleton. If the tissue is undergoing tension, this cut will induce a recoil in the surrounding regions of the cut. This protocol describes how one can perform laser microdissection experiments and subsequently measure the recoil speed of the sample of interest. While we explain how to carry out these experiments in Drosophila embryos, the recoil calibration and downstream analyses can be applied to other types of preparations. Key features Allows measuring tension in live Drosophila embryos with a relatively simple approach. Describes a quick way to mount a high number of embryos. Includes a segmentation-free recoil quantification that reduces bias and speeds up analysis. Graphical overview.