title: Effects of Noninhibitory Serpin Maspin on the Actin Cytoskeleton: A Quantitative Image Modeling Approach.
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.
authors.affiliation: 1Computational Intelligence Group, Faculty of Engineering and Environment,Northumbria University,Newcastle upon Tyne NE1 8ST,UK.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Al-Mamun
authors.affiliation: 3School of Biological Sciences,University of East Anglia,Norwich,Norfolk, NR4 7TJ,UK.
authors.forename: Lorna
authors.initials: L
authors.lastname: Ravenhill
authors.affiliation: 1Computational Intelligence Group, Faculty of Engineering and Environment,Northumbria University,Newcastle upon Tyne NE1 8ST,UK.
authors.forename: Worawut
authors.initials: W
authors.lastname: Srisukkham
authors.affiliation: 1Computational Intelligence Group, Faculty of Engineering and Environment,Northumbria University,Newcastle upon Tyne NE1 8ST,UK.
authors.forename: Alamgir
authors.initials: A
authors.lastname: Hossain
authors.affiliation: 1Computational Intelligence Group, Faculty of Engineering and Environment,Northumbria University,Newcastle upon Tyne NE1 8ST,UK.
authors.forename: Charles
authors.initials: C
authors.lastname: Fall
authors.affiliation: 3School of Biological Sciences,University of East Anglia,Norwich,Norfolk, NR4 7TJ,UK.
authors.forename: Vincent
authors.initials: V
authors.lastname: Ellis
authors.affiliation: 5Department of Applied Sciences, Faculty of Health and Life Sciences,Northumbria University,Newcastle upon Tyne NE1 8ST,UK.
authors.forename: Rosemary
authors.initials: R
authors.lastname: Bass
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects* / Q000648:ultrastructure*; D045744:Cell Line, Tumor; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D018613:Microscopy, Confocal; D015843:Serpins / Q000378:metabolism*
chemical_list: C085344:SERPIN-B5; D015843:Serpins
keywords: cell migration; quantitative image modeling; serpinB5; watershed segmentation