scholarly journals Epithelial geometry regulates spindle orientation and progenitor fate during formation of the mammalian epidermis

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Kimberly Box ◽  
Bradley W Joyce ◽  
Danelle Devenport
Keyword(s):  
Cell Fate ◽  
Planar Cell Polarity ◽  
Time Lapse ◽  
Mouse Embryos ◽  
Spindle Orientation ◽  
Basal Cells ◽  
Tissue Architecture ◽  
Cell Geometry ◽  
Oriented Cell Division ◽  
Time Lapse Imaging

The control of cell fate through oriented cell division is imperative for proper organ development. Basal epidermal progenitor cells divide parallel or perpendicular to the basement membrane to self-renew or produce differentiated stratified layers, but the mechanisms regulating the choice between division orientations are unknown. Using time-lapse imaging to follow divisions and fates of basal progenitors, we find that mouse embryos defective for the planar cell polarity (PCP) gene, Vangl2, exhibit increased perpendicular divisions and hyperthickened epidermis. Surprisingly, this is not due to defective Vangl2 function in the epidermis, but to changes in cell geometry and packing that arise from the open neural tube characteristic of PCP mutants. Through regional variations in epidermal deformation and physical manipulations, we show that local tissue architecture, rather than cortical PCP cues, regulates the decision between symmetric and stratifying divisions, allowing flexibility for basal cells to adapt to the needs of the developing tissue.

scholarly journals Time-Lapse Imaging of Postimplantation Mouse Embryos

2011 ◽  
Vol 2011 (4) ◽  
pp. pdb.prot5595-pdb.prot5595 ◽  
Author(s):  
M. D. Garcia ◽  
R. S. Udan ◽  
A.-K. Hadjantonakis ◽  
M. E. Dickinson
Keyword(s):  
Time Lapse ◽  
Mouse Embryos ◽  
Time Lapse Imaging

scholarly journals Microwells support high-resolution time-lapse imaging and development of preimplanted mouse embryos

2015 ◽  
Vol 9 (2) ◽  
pp. 022407 ◽  
Author(s):  
Yu-Hsiang Chung ◽  
Yi-Hsing Hsiao ◽  
Wei-Lun Kao ◽  
Chia-Hsien Hsu ◽  
Da-Jeng Yao ◽  
...  
Keyword(s):  
High Resolution ◽  
Time Lapse ◽  
Mouse Embryos ◽  
Resolution Time ◽  
Time Lapse Imaging

scholarly journals Planar cell polarity signaling coordinates oriented cell division and cell rearrangement in clonally expanding growth plate cartilage

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Yuwei Li ◽  
Ang Li ◽  
Jason Junge ◽  
Marianne Bronner
Keyword(s):  
Cell Division ◽  
Cell Polarity ◽  
Planar Cell Polarity ◽  
Cleavage Furrow ◽  
Tissue Architecture ◽  
Oriented Cell Division ◽  
Cellular Events ◽  
Column Formation ◽  
Single Column ◽  
Sister Cells

Both oriented cell divisions and cell rearrangements are critical for proper embryogenesis and organogenesis. However, little is known about how these two cellular events are integrated. Here we examine the linkage between these processes in chick limb cartilage. By combining retroviral-based multicolor clonal analysis with live imaging, the results show that single chondrocyte precursors can generate both single-column and multi-column clones through oriented division followed by cell rearrangements. Focusing on single column formation, we show that this stereotypical tissue architecture is established by a pivot-like process between sister cells. After mediolateral cell division, N-cadherin is enriched in the post-cleavage furrow; then one cell pivots around the other, resulting in stacking into a column. Perturbation analyses demonstrate that planar cell polarity signaling enables cells to pivot in the direction of limb elongation via this N-cadherin-mediated coupling. Our work provides new insights into the mechanisms generating appropriate tissue architecture of limb skeleton.

scholarly journals Junctional tumor suppressors interact with 14-3-3 proteins to control planar spindle alignment

2019 ◽  
Vol 218 (6) ◽  
pp. 1824-1838 ◽  
Author(s):  
Yu-ichiro Nakajima ◽  
Zachary T. Lee ◽  
Sean A. McKinney ◽  
Selene K. Swanson ◽  
Laurence Florens ◽  
...  
Keyword(s):  
Cell Fate ◽  
Tumor Suppressors ◽  
Mitotic Spindle ◽  
Wing Disc ◽  
Spindle Orientation ◽  
Epithelial Proliferation ◽  
Tissue Architecture ◽  
Drosophila Wing ◽  
Discs Large

Proper orientation of the mitotic spindle is essential for cell fate determination, tissue morphogenesis, and homeostasis. During epithelial proliferation, planar spindle alignment ensures the maintenance of polarized tissue architecture, and aberrant spindle orientation can disrupt epithelial integrity. Nevertheless, in vivo mechanisms that restrict the mitotic spindle to the plane of the epithelium remain poorly understood. Here we show that the junction-localized tumor suppressors Scribbled (Scrib) and Discs large (Dlg) control planar spindle orientation via Mud and 14-3-3 proteins in the Drosophila wing disc epithelium. During mitosis, Scrib is required for the junctional localization of Dlg, and both affect mitotic spindle movements. Using coimmunoprecipitation and mass spectrometry, we identify 14-3-3 proteins as Dlg-interacting partners and further report that loss of 14-3-3s causes both abnormal spindle orientation and disruption of epithelial architecture as a consequence of basal cell delamination and apoptosis. Combined, these biochemical and genetic analyses indicate that 14-3-3s function together with Scrib, Dlg, and Mud during planar cell division.

scholarly journals Multiplexed End-point Microfluidic Chemotaxis Assay using Centrifugal Alignment

2020 ◽  
Author(s):  
Sampath Satti ◽  
Pan Deng ◽  
Kerryn Matthews ◽  
Simon P. Duffy ◽  
Hongshen Ma
Keyword(s):  
Fluid Flow ◽  
Drug Testing ◽  
Time Lapse ◽  
Cell Alignment ◽  
Cell Geometry ◽  
End Point ◽  
Starting Point ◽  
Correct Location ◽  
Set Up ◽  
Time Lapse Imaging

AbstractA fundamental challenge to multiplexing microfluidic chemotaxis assays at scale is the requirement for time-lapse imaging to continuously track migrating cells. Drug testing and drug screening applications require the ability to perform hundreds of experiments in parallel, which is not feasible for assays that require continuous imaging. To address this limitation, end-point chemotaxis assays have been developed using fluid flow to align cells in traps or sieves prior to cell migration. However, these methods require precisely controlled fluid flow to transport cells to the correct location without undesirable mechanical stress, which introduce significant set up time and design complexity. Here, we describe a microfluidic device that eliminates the need for precise flow control by using centrifugation to align cells at a common starting point. A chemoattractant gradient is then formed using passive diffusion prior to chemotaxis in an incubated environment. This approach provides a simple and scalable approach to multiplexed chemotaxis assays. Centrifugal alignment is also insensitive to cell geometry, enabling this approach to be compatible with primary cell samples that are often heterogeneous. We demonstrate the capability of this approach by assessing chemotaxis of primary neutrophils in response to an fMLP (N-formyl-met-leu-phe) gradient. Our results show that cell alignment by centrifugation offers a potential avenue to develop scalable end-point multiplexed microfluidic chemotaxis assays.

scholarly journals Hybrid optical gating for long-term 3D time-lapse imaging of the beating embryonic zebrafish heart

2019 ◽  
Author(s):  
Jonathan M. Taylor ◽  
Carl J. Nelson ◽  
Finnius A. Bruton ◽  
Aryan K. Baghbadrani ◽  
Charlotte Buckley ◽  
...  
Keyword(s):  
Cell Fate ◽  
Cardiac Cycle ◽  
Physiological State ◽  
Time Lapse ◽  
Computational Techniques ◽  
Phase Lock ◽  
Optical Gating ◽  
Time Lapse Imaging ◽  
Zebrafish Heart

AbstractThree-dimensional fluorescence time-lapse imaging of structural, cellular and sub-cellular processes in the beating heart is an increasingly achievable goal using the latest imaging and computational techniques. However, previous approaches have had significant limitations. Temporarily arresting the heart using drugs disrupts the heart’s physiological state, and the use of ultra-high frame-rates for fluorescence image acquisition causes phototoxic cell damage. Real-time triggered imaging, synchronized to a specific phase in the cardiac-cycle, can computationally “freeze” the heart to acquire the minimal number of fluorescence images required for 3D time-lapse imaging. However, until now no solution has been able to maintain phase-lock to the same point in the cardiac cycle for more than about one hour. Our new hybrid optical gating system maintains phase-lock for up to 24 h, acquiring synchronized 3D+time video stacks of the unperturbed heart in vivo. This approach has enabled us to observe detailed developmental, structural, cellular and subcellular processes, including live cell division and cell fate tracking, in the embryonic zebrafish heart using transgenic fish lines expressing cell-specific fluorophores. We show that our approach not only provides high spatial and temporal resolution 3D-imaging, but also avoids phototoxic injury, where alternative approaches induce measurable harm. This provides superb cellular and subcellular imaging of the heart while it is beating in its normal physiological state, and opens up new and exciting opportunities for further study in the heart and other moving cellular and subcellular structures in vivo.

scholarly journals Time-Lapse Imaging of Neuroblastoma Cells to Determine Cell Fate upon Gene Knockdown

PLoS ONE ◽  
2012 ◽  
Vol 7 (12) ◽  
pp. e50988 ◽  
Author(s):  
Richa Batra ◽  
Nathalie Harder ◽  
Sina Gogolin ◽  
Nicolle Diessl ◽  
Zita Soons ◽  
...  
Keyword(s):  
Cell Fate ◽  
Neuroblastoma Cells ◽  
Time Lapse ◽  
Gene Knockdown ◽  
Time Lapse Imaging

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.

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