Cellular and Nuclear Alignment Analysis for Determining Epithelial Cell Chirality

Raymond, Michael; Ray, Poulomi; Kaur, Gurleem; Singh, Ajay Vikram; Wan, Leo Q.

doi:10.1007/s10439-015-1431-3

Cited by 36 publications

(41 citation statements)

References 30 publications

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“…Among four quadrants defined by circumferential and radial axes, two (−180° – −90° and 0° – 90°) were defined as counterclockwise (CCW) alignment and two (−90° – 0° and 90° – 180°) as clockwise (CW) alignment, following the convention outlined before 47 . Consistently with previous analyses 29 , both cell and nucleus based analyses had a significant bias towards counterclockwise (CCW) alignment among chiral rings.…”

Section: Resultssupporting

confidence: 87%

“…We used a Python-based automated object-labeling connected-component method to analyze individual cells and their nuclei, as reported before 29 . Briefly, ZO-1 images were adjusted for brightness, and the background was subtracted to highlight the cell edges and to minimize artifacts in ImageJ.…”

Section: Methodsmentioning

confidence: 99%

“…None of these analyses, however, were based on individual cells, and the physical nature behind cell chirality (i.e., the coordination of multiaxial polarity) was largely ignored. Here, we utilized a recently developed Python-based algorithm to evaluate the polarizations of individual cells, nuclei and their relative positioning within a geometrically controlled cellular monolayer 29 . Such an approach enables accurate and robust analyses of epithelial chiral morphogenesis.…”

Section: Introductionmentioning

confidence: 99%

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Multiaxial Polarity Determines Individual Cellular and Nuclear Chirality

et al. 2016

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Intrinsic cell chirality has been implicated in the left-right (LR) asymmetry of embryonic development. Impaired cell chirality could lead to severe birth defects in laterality. Previously, we detected cell chirality with an in vitro micropatterning system. Here, we demonstrate for the first time that chirality can be quantified as the coordination of multiaxial polarization of individual cells and nuclei. Using an object labeling, connected component based method, we characterized cell chirality based on cell and nuclear shape polarization and nuclear positioning of each cell in multicellular patterns of epithelial cells. We found that the cells adopted a LR bias the boundaries by positioning the sharp end towards the leading edge and leaving the nucleus at the rear. This behavior is consistent with the directional migration observed previously on the boundary of micropatterns. Although the nucleus is chirally aligned, it is not strongly biased towards or away from the boundary. As the result of the rear positioning of nuclei, the nuclear positioning has an opposite chirality to that of cell alignment. Overall, our results have revealed deep insights of chiral morphogenesis as the coordination of multiaxial polarization at the cellular and subcellular levels.

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Section: Resultssupporting

confidence: 87%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Multiaxial Polarity Determines Individual Cellular and Nuclear Chirality

et al. 2016

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“…Whether the handedness is dextral or sinistral depends on the cell line [49]. Chirality in the nuclear shape and the involvement of E-cadherin in transmitting a chiral bias to neighbouring cells were shown using Madin–Darby canine kidney epithelial cells [51,52]. …”

Section: Cell Chirality In Vertebrate Cultured Cellsmentioning

confidence: 99%

Cell chirality: its origin and roles in left–right asymmetric development

Inaki

Liu

Matsuno

2016

Phil. Trans. R. Soc. B

144

147

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An item is chiral if it cannot be superimposed on its mirror image. Most biological molecules are chiral. The homochirality of amino acids ensures that proteins are chiral, which is essential for their functions. Chirality also occurs at the whole-cell level, which was first studied mostly in ciliates, single-celled protozoans. Ciliates show chirality in their cortical structures, which is not determined by genetics, but by ‘cortical inheritance’. These studies suggested that molecular chirality directs whole-cell chirality. Intriguingly, chirality in cellular structures and functions is also found in metazoans. In Drosophila, intrinsic cell chirality is observed in various left–right (LR) asymmetric tissues, and appears to be responsible for their LR asymmetric morphogenesis. In other invertebrates, such as snails and Caenorhabditis elegans, blastomere chirality is responsible for subsequent LR asymmetric development. Various cultured cells of vertebrates also show intrinsic chirality in their cellular behaviours and intracellular structural dynamics. Thus, cell chirality may be a general property of eukaryotic cells. In Drosophila, cell chirality drives the LR asymmetric development of individual organs, without establishing the LR axis of the whole embryo. Considering that organ-intrinsic LR asymmetry is also reported in vertebrates, this mechanism may contribute to LR asymmetric development across phyla.This article is part of the themed issue ‘Provocative questions in left–right asymmetry’.

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“…In conventional approaches, fluorescence-stained nuclei are used to reveal cellular orientation (Raymond, Ray, Kaur, Singh, & Wan, 2016), and a global threshold is commonly used to binarize the images for further analysis. Unfortunately, in a culture with high cell density, densely packed cell cluster makes the contours of nuclei overlapping.…”

mentioning

confidence: 99%

Outline‐etching image segmentation reveals enhanced cell chirality through intercellular alignment

Huang

Bao

Kwong

et al. 2018

Biotech & Bioengineering

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Cells cultured on micropatterns exhibit a chiral orientation, which may underlie the development of left–right asymmetry in tissue microarchitectures. To investigate this phenomenon, fluorescence staining of nuclei has been used to reveal such orientation. However, for images with high cell density, analysis is difficult because of the overlapping nuclei. Here, we report an image processing method that can acquire cell orientations within dense cell populations. After initial separation based on Boolean addition of binarized images using global and adaptive thresholds, the overlapping nucleus contours in the binarized images were segmented by iteratively etching the outlines of nuclei, which allowed the orientations of each cell to be extracted from densely packed cell clusters. In applying this technique to cultured C2C12 myoblasts in micropatterned stripes on different substrates, we found an enhanced chiral orientation on glass substrate. More important, this enhanced chirality was consistently observed with increased intercellular alignment and independent of cell–cell distance or cell density, suggesting that intercellular alignment plays a role in determining the chiral orientation. By segmenting single cells with intact orientation, this technique offers an automated method for quantitative analysis with improved accuracy, providing an essential tool for studying left–right asymmetry and other morphogenic dynamics in tissue formation.

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Cellular and Nuclear Alignment Analysis for Determining Epithelial Cell Chirality

Cited by 36 publications

References 30 publications

Multiaxial Polarity Determines Individual Cellular and Nuclear Chirality

Multiaxial Polarity Determines Individual Cellular and Nuclear Chirality

Cell chirality: its origin and roles in left–right asymmetric development

Outline‐etching image segmentation reveals enhanced cell chirality through intercellular alignment

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