scholarly journals Trpml controls actomyosin contractility and couples migration to phagocytosis in fly macrophages

2020 ◽  
Vol 219 (3) ◽  
Author(s):  
Sandra Sofía Edwards-Jorquera ◽  
Floris Bosveld ◽  
Yohanns A. Bellaïche ◽  
Ana-María Lennon-Duménil ◽  
Álvaro Glavic
Keyword(s):  
Cell Migration ◽  
Immune Cells ◽  
Space Exploration ◽  
Time Lapse ◽  
Extracellular Material ◽  
Cell Locomotion ◽  
Actomyosin Contractility ◽  
Time Lapse Imaging ◽  
Actomyosin Cytoskeleton

Phagocytes use their actomyosin cytoskeleton to migrate as well as to probe their environment by phagocytosis or macropinocytosis. Although migration and extracellular material uptake have been shown to be coupled in some immune cells, the mechanisms involved in such coupling are largely unknown. By combining time-lapse imaging with genetics, we here identify the lysosomal Ca2+ channel Trpml as an essential player in the coupling of cell locomotion and phagocytosis in hemocytes, the Drosophila macrophage-like immune cells. Trpml is needed for both hemocyte migration and phagocytic processing at distinct subcellular localizations: Trpml regulates hemocyte migration by controlling actomyosin contractility at the cell rear, whereas its role in phagocytic processing lies near the phagocytic cup in a myosin-independent fashion. We further highlight that Vamp7 also regulates phagocytic processing and locomotion but uses pathways distinct from those of Trpml. Our results suggest that multiple mechanisms may have emerged during evolution to couple phagocytic processing to cell migration and facilitate space exploration by immune cells.

scholarly journals Migration of immortalized nasopharyngeal epithelia and carcinoma cells through porous membrane in 3D platforms

2020 ◽  
Vol 40 (6) ◽  
Author(s):  
Ziyu Liu ◽  
Weiguan Zhang ◽  
Stella W. Pang
Keyword(s):  
Cell Migration ◽  
Nasopharyngeal Carcinoma ◽  
Cell Line ◽  
Time Lapse ◽  
Porous Membrane ◽  
Epithelial Cell Line ◽  
3D Microenvironment ◽  
Porous Interface ◽  
Time Lapse Imaging

Abstract In the present study, 3D biomimetic platforms were fabricated with guiding grating to mimic extracellular matrix topography, porous membrane to resemble the epithelial porous interface and trenches below to represent blood vessels as an in vitro tissue microenvironment. Fabrication technologies were developed to integrate the transparent biocompatible polydimethylsiloxane platforms with preciously controlled dimensions. Cell migration behaviors of an immortalized nasopharyngeal epithelial cell line (NP460) and a nasopharyngeal carcinoma cell line (NPC43) were studied on the 2D and 3D platforms. The NP460 and NPC43 cells traversing through the porous membrane and migrating in the trenches below were studied by time-lapse imaging. Before traversing through the pores, NP460 and NPC43 cells migrated around the pores but NPC43 cells had a lower migration speed with less lamellipodia spreading. After traversing to trenches below, NPC43 cells moved faster with an alternated elongated morphology (mesenchymal migration mode) and round morphology (amoeboid migration mode) compared with only mesenchymal migration mode for NP460 cells. The cell traversing probability through porous membrane on platforms with 30 μm wide trenches below was found to be the highest when the guiding grating was perpendicular to the trenches below and the lowest when the guiding grating was parallel to the trenches below. The present study shows important information on cell migration in complex 3D microenvironment with various dimensions and could provide insight for pathology and treatment of nasopharyngeal carcinoma.

scholarly journals Endoscopic Time-Lapse Imaging of Immune Cells in Infarcted Mouse Hearts

2013 ◽  
Vol 112 (6) ◽  
pp. 891-899 ◽  
Author(s):  
Keehoon Jung ◽  
Pilhan Kim ◽  
Florian Leuschner ◽  
Rostic Gorbatov ◽  
Jun Ki Kim ◽  
...  
Keyword(s):  
Immune Cells ◽  
Time Lapse ◽  
Time Lapse Imaging

scholarly journals A versatile aquaporin-2 cell system for quantitative temporal expression and live cell imaging

2019 ◽  
Vol 317 (1) ◽  
pp. F124-F132 ◽  
Author(s):  
Mikkel R. Holst ◽  
Lene N. Nejsum
Keyword(s):  
Plasma Membrane ◽  
Cell Migration ◽  
Fluorescent Protein ◽  
Time Lapse ◽  
Cell System ◽  
Urine Concentration ◽  
Epithelial Morphogenesis ◽  
Expression Levels ◽  
Aquaporin 2 ◽  
Time Lapse Imaging

Aquaporin-2 (AQP2) fine tunes urine concentration in response to the antidiuretic hormone vasopressin. In addition, AQP2 has been suggested to promote cell migration and epithelial morphogenesis. A cell system allowing temporal and quantitative control of expression levels of AQP2 and phospho-mimicking mutants has been missing, as has a system allowing expression of fluorescently tagged AQP2 for time-lapse imaging. In the present study, we generated and validated a Flp-In T-REx Madin-Darby canine kidney cell system for temporal and quantitative control of AQP2 and phospho-mimicking mutants. We verified that expression levels can be temporally and quantitatively controlled and that AQP2 translocated to the plasma membrane in response to elevated cAMP, which also induced S256 phosphorylation. The phospho-mimicking mutants AQP2-S256A and AQP2-S256D localized as previously described, primarily intracellular and to the plasma membrane, respectively. Induction of AQP2 expression in combination with transient, low expression of enhanced green fluorescent protein-tagged AQP2 enabled expression without aggregation and correct translocation in response to elevated cAMP. Interestingly, time-lapse imaging revealed AQP2-containing tubulating endosomes and that tubulation significantly decreased 30 min after cAMP elevation. This was mirrored by the phospho-mimicking mutants AQP2-S256A and AQP2-S256D, where AQP2-S256A-containing endosomes tubulated, whereas AQP2-S256D-containing endosomes did not. Thus, this cell system enables a multitude of cell-based assays warranted to provide deeper insights into the mechanisms of AQP2 regulation and effects on cell migration and epithelial morphogenesis.

scholarly journals The dynamic interplay between ATP/ADP levels and autophagy sustain neuronal migration in vivo

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Cedric Bressan ◽  
Alessandra Pecora ◽  
Dave Gagnon ◽  
Marina Snapyan ◽  
Simon Labrecque ◽  
...  
Keyword(s):  
Cell Migration ◽  
Stationary Phase ◽  
Neuronal Migration ◽  
Stationary Phases ◽  
Focal Adhesions ◽  
Time Lapse ◽  
Time Lapse Imaging ◽  
Migratory Stream ◽  
Migrating Cells

Cell migration is a dynamic process that entails extensive protein synthesis and recycling, structural remodeling, and considerable bioenergetic demand. Autophagy is one of the pathways that maintain cellular homeostasis. Time-lapse imaging of autophagosomes and ATP/ADP levels in migrating cells in the rostral migratory stream of mouse revealed that decreases in ATP levels force cells into the stationary phase and induce autophagy. Pharmacological or genetic impairments of autophagy in neuroblasts using either bafilomycin, inducible conditional mice, or CRISPR/Cas9 gene editing decreased cell migration due to the longer duration of the stationary phase. Autophagy is modulated in response to migration-promoting and inhibiting molecular cues and is required for the recycling of focal adhesions. Our results show that autophagy and energy consumption act in concert in migrating cells to dynamically regulate the pace and periodicity of the migratory and stationary phases to sustain neuronal migration.

Cleavage of Human Embryos: Options and Diversity

Acta Naturae ◽  
2016 ◽  
Vol 8 (3) ◽  
pp. 88-96
Author(s):  
Yu. K. Doronin ◽  
I. V. Senechkin ◽  
L. V. Hilkevich ◽  
M. A. Kurcer
Keyword(s):  
Time Lapse ◽  
Reproductive Technologies ◽  
Human Embryos ◽  
Imaging Data ◽  
Noninvasive Methods ◽  
Topographic Features ◽  
Time Parameters ◽  
Embryo Cleavage ◽  
Time Lapse Imaging ◽  
Developmental Dynamics

In order to estimate the diversity of embryo cleavage relatives to embryo progress (blastocyst formation), time-lapse imaging data of preimplantation human embryo development were used. This retrospective study is focused on the topographic features and time parameters of the cleavages, with particular emphasis on the lengths of cleavage cycles and the genealogy of blastomeres in 2- to 8-cell human embryos. We have found that all 4-cell human embryos have four developmental variants that are based on the sequence of appearance and orientation of cleavage planes during embryo cleavage from 2 to 4 blastomeres. Each variant of cleavage shows a strong correlation with further developmental dynamics of the embryos (different cleavage cycle characteristics as well as lengths of blastomere cycles). An analysis of the sequence of human blastomere divisions allowed us to postulate that the effects of zygotic determinants are eliminated as a result of cleavage, and that, thereafter, blastomeres acquire the ability of own syntheses, regulation, polarization, formation of functional contacts, and, finally, of specific differentiation. This data on the early development of human embryos obtained using noninvasive methods complements and extend our understanding of the embryogenesis of eutherian mammals and may be applied in the practice of reproductive technologies.

scholarly journals Evaluating the utility of time-lapse imaging in the estimation of post-mortem interval: An Australian case study

2019 ◽  
Vol 1 ◽  
pp. 204-210 ◽  
Author(s):  
Alyson Wilson ◽  
Stanley Serafin ◽  
Dilan Seckiner ◽  
Rachel Berry ◽  
Xanthé Mallett
Keyword(s):  
Time Lapse ◽  
Post Mortem ◽  
Post Mortem Interval ◽  
Australian Case ◽  
Time Lapse Imaging
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