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Effects of Noninhibitory Serpin Maspin on the Actin Cytoskeleton: A Quantitative Image Modeling Approach

Published online by Cambridge University Press:  24 February 2016

Mohammed Al-Mamun
Affiliation:
Computational Intelligence Group, Faculty of Engineering and Environment, Northumbria University, Newcastle upon Tyne NE1 8ST, UK Department of Population Medicine & Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850, USA
Lorna Ravenhill
Affiliation:
School of Biological Sciences, University of East Anglia, Norwich, Norfolk, NR4 7TJ, UK
Worawut Srisukkham
Affiliation:
Computational Intelligence Group, Faculty of Engineering and Environment, Northumbria University, Newcastle upon Tyne NE1 8ST, UK
Alamgir Hossain
Affiliation:
Computational Intelligence Group, Faculty of Engineering and Environment, Northumbria University, Newcastle upon Tyne NE1 8ST, UK Anglia Ruskin IT Research Institute (ARITI), Anglia Ruskin University, Cambridge CB1 1PT, UK
Charles Fall
Affiliation:
Computational Intelligence Group, Faculty of Engineering and Environment, Northumbria University, Newcastle upon Tyne NE1 8ST, UK
Vincent Ellis
Affiliation:
School of Biological Sciences, University of East Anglia, Norwich, Norfolk, NR4 7TJ, UK
Rosemary Bass*
Affiliation:
Department of Applied Sciences, Faculty of Health and Life Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK
*
* Corresponding author. rosemary.bass@northumbria.ac.uk

Abstract

Recent developments in quantitative image analysis allow us to interrogate confocal microscopy images to answer biological questions. Clumped and layered cell nuclei and cytoplasm in confocal images challenges the ability to identify subcellular compartments. To date, there is no perfect image analysis method to identify cytoskeletal changes in confocal images. Here, we present a multidisciplinary study where an image analysis model was developed to allow quantitative measurements of changes in the cytoskeleton of cells with different maspin exposure. Maspin, a noninhibitory serpin influences cell migration, adhesion, invasion, proliferation, and apoptosis in ways that are consistent with its identification as a tumor metastasis suppressor. Using different cell types, we tested the hypothesis that reduction in cell migration by maspin would be reflected in the architecture of the actin cytoskeleton. A hybrid marker-controlled watershed segmentation technique was used to segment the nuclei, cytoplasm, and ruffling regions before measuring cytoskeletal changes. This was informed by immunohistochemical staining of cells transfected stably or transiently with maspin proteins, or with added bioactive peptides or protein. Image analysis results showed that the effects of maspin were mirrored by effects on cell architecture, in a way that could be described quantitatively.

Type
Biological Applications
Copyright
© Microscopy Society of America 2016 

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Footnotes

a

These authors contributed equally.

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Effects of Noninhibitory Serpin Maspin on the Actin Cytoskeleton: A Quantitative Image Modeling Approach
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