Effect of Cell Shape and Dimensionality on Spindle Orientation and Mitotic Timing
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{"title"=>"Effect of Cell Shape and Dimensionality on Spindle Orientation and Mitotic Timing", "type"=>"journal", "authors"=>[{"first_name"=>"Mirren", "last_name"=>"Charnley", "scopus_author_id"=>"23977785700"}, {"first_name"=>"Fabian", "last_name"=>"Anderegg", "scopus_author_id"=>"12241323700"}, {"first_name"=>"René", "last_name"=>"Holtackers", "scopus_author_id"=>"8639424600"}, {"first_name"=>"Marcus", "last_name"=>"Textor", "scopus_author_id"=>"7007007759"}, {"first_name"=>"Patrick", "last_name"=>"Meraldi", "scopus_author_id"=>"6602878317"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84879173972", "pui"=>"369142456", "doi"=>"10.1371/journal.pone.0066918", "sgr"=>"84879173972", "pmid"=>"23825020"}, "id"=>"375c47b8-c751-3d97-b535-455e76f6e31f", "abstract"=>"The formation and orientation of the mitotic spindle is a critical feature of mitosis. The morphology of the cell and the spatial distribution and composition of the cells' adhesive microenvironment all contribute to dictate the position of the spindle. However, the impact of the dimensionality of the cells' microenvironment has rarely been studied. In this study we present the use of a microwell platform, where the internal surfaces of the individual wells are coated with fibronectin, enabling the three-dimensional presentation of adhesive ligands to single cells cultured within the microwells. This platform was used to assess the effect of dimensionality and cell shape in a controlled microenvironment. Single HeLa cells cultured in circular microwells exhibited greater tilting of the mitotic spindle, in comparison to cells cultured in square microwells. This correlated with an increase in the time required to align the chromosomes at the metaphase plate due to prolonged activation of the spindle checkpoint in an actin dependent process. The comparison to 2D square patterns revealed that the dimensionality of cell adhesions alone affected both mitotic timings and spindle orientation; in particular the role of actin varied according to the dimensionality of the cells' microenvironment. Together, our data revealed that cell shape and the dimensionality of the cells' adhesive environment impacted on both the orientation of the mitotic spindle and progression through mitosis.", "link"=>"http://www.mendeley.com/research/effect-cell-shape-dimensionality-spindle-orientation-mitotic-timing", "reader_count"=>37, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>15, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>9, "Student > Master"=>4, "Other"=>1, "Student > Bachelor"=>3, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>15, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>9, "Student > Master"=>4, "Other"=>1, "Student > Bachelor"=>3, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>9, "Materials Science"=>1, "Agricultural and Biological Sciences"=>20, "Philosophy"=>1, "Neuroscience"=>1, "Physics and Astronomy"=>3}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Materials Science"=>{"Materials Science"=>1}, "Neuroscience"=>{"Neuroscience"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>20}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>9}, "Unspecified"=>{"Unspecified"=>1}, "Philosophy"=>{"Philosophy"=>1}}, "reader_count_by_country"=>{"United States"=>1, "United Kingdom"=>1, "France"=>2}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["http://files.figshare.com/1090987/Figure_1.tif"], "pos_in_sequence"=>0, "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "users"=>["Fabian Anderegg", "Marcus Textor", "Patrick Meraldi", "René Holtackers", "Mirren Charnley"], "links"=>[], "tags"=>["Extracellular matrix", "Biological systems engineering", "3d", "mitosis", "biotechnology", "Cell physiology", "Musculoskeletal system", "cell adhesion", "cell division", "Bioengineering", "microwell", "Anatomy and physiology", "biomechanics", "Cell mechanics", "Molecular cell biology", "Extracellular matrix adhesions"], "title"=>"<p>Control of 3D cell shape using a microwell cell culture platform.</p>", "figshare_url"=>"http://figshare.com/articles/_Control_of_3D_cell_shape_using_a_microwell_cell_culture_platform_/723544", "defined_type"=>1, "doi"=>["http://dx.doi.org/10.1371/journal.pone.0066918.g001"], "published_date"=>"2013-06-18 00:59:04", "article_id"=>723544, "categories"=>["Biological Sciences", "Engineering"], "description"=>"<p >(A&#8211;B) HeLa cells (RFP-tubulin/GFP-H2B) were synchronized and cultured in square (A) or circular (B) microwells for 18 hours and imaged for nucleus (green) and cell outline (Vybrant DiD cell labeling solution, red). Images show the central xy slice of the cell (top) and the corresponding xz slice of the same cell (bottom); bars: 10 &#181;m.</p>"}
  • {"files"=>["http://files.figshare.com/1090988/Figure_2.tif"], "pos_in_sequence"=>0, "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "users"=>["Fabian Anderegg", "Marcus Textor", "Patrick Meraldi", "René Holtackers", "Mirren Charnley"], "links"=>[], "tags"=>["Extracellular matrix", "Biological systems engineering", "mitosis", "biotechnology", "Cell physiology", "Musculoskeletal system", "cell adhesion", "cell division", "Bioengineering", "Anatomy and physiology", "biomechanics", "Cell mechanics", "Molecular cell biology", "mitotic", "Extracellular matrix adhesions"], "title"=>"<p>Effect of cell shape on the orientation of the mitotic spindle.</p>", "figshare_url"=>"http://figshare.com/articles/_Effect_of_cell_shape_on_the_orientation_of_the_mitotic_spindle_/723545", "defined_type"=>1, "doi"=>["http://dx.doi.org/10.1371/journal.pone.0066918.g002"], "published_date"=>"2013-06-18 00:59:05", "article_id"=>723545, "categories"=>["Biological Sciences", "Engineering"], "description"=>"<p >The angle of mitotic spindle at metaphase was assessed in HeLa (GFP-H2B/RFP-tubulin or GFP-centrin-2/RFP-tubulin) cells cultured on the different platforms. To determine the orientation of the spindle parallel to the substrate plane, cells were imaged for microwell and cell shape (transmission channel, blue), DNA (green) and tubulin (red) after culture in (A) square microwells or (B) circular microwells; bars: 10 &#181;m. Cells cultured (C) on 2D substrates or (E) within circular microwells possessed a random orientation of the mitotic spindle. (D) Conversely, a high proportion of the cells cultured in square microwells aligned the spindle along the long axis of the cell. To determine the orientation of the spindle perpendicular to the substrate plane, z stacks were reconstructed to obtain the xz view of the cell. Representative images are shown of cells with a (F) parallel versus (G) tilted orientation of the mitotic spindle; bars: 10 &#181;m. (H) The angle of the spindle was dependent on whether cells were cultured on 2D homogenously coated substrates (black bars), or within square (light grey bars) or circular (dark grey bars) microwells. A higher percentage of cells cultured in circular microwells exhibited tilting of the mitotic spindle, in comparison to cells cultured in square microwells and on 2D substrates. (I) Furthermore, on average greater tilting was observed after cells were cultured in the circular microwells, while the mitotic spindle was aligned nearly parallel to the substrate in cells cultured in 2D or square microwells. Values represent spindle angle in degrees &#177; SEM. Key: *** p&lt;0.001.</p>"}
  • {"files"=>["http://files.figshare.com/1090989/Figure_3.tif"], "pos_in_sequence"=>0, "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "users"=>["Fabian Anderegg", "Marcus Textor", "Patrick Meraldi", "René Holtackers", "Mirren Charnley"], "links"=>[], "tags"=>["Extracellular matrix", "Biological systems engineering", "progression", "mitosis", "biotechnology", "Cell physiology", "Musculoskeletal system", "cell adhesion", "cell division", "Bioengineering", "Anatomy and physiology", "biomechanics", "Cell mechanics", "Molecular cell biology", "Extracellular matrix adhesions"], "title"=>"<p>The effect of cell shape on the cells' progression through mitosis.</p>", "figshare_url"=>"http://figshare.com/articles/_The_effect_of_cell_shape_on_the_cells_progression_through_mitosis_/723546", "defined_type"=>1, "doi"=>["http://dx.doi.org/10.1371/journal.pone.0066918.g003"], "published_date"=>"2013-06-18 00:59:06", "article_id"=>723546, "categories"=>["Biological Sciences", "Engineering"], "description"=>"<p >HeLa cells (RFP-tubulin/GFP-H2B) were synchronized and cultured on 2D homogenously coated substrates (black bars) or within square (light grey bars) or circular (dark grey bars) microwells and assessed for the time required to progress through each stage in mitosis. Cells cultured in circular microwells took longer to progress through mitosis and specifically to reach late prometaphase, than cells cultured in square microwells or on 2D substrates. All values represent time in minutes &#177; SEM. Key: * p&lt;0.05, ** p&lt;0.01; NB &#8202;=&#8202; nuclear envelope breakdown, LPM &#8202;=&#8202; late prometaphase, M&#8202;=&#8202; metaphase, A&#8202;=&#8202; anaphase, C&#8202;=&#8202; cytokinesis.</p>"}
  • {"files"=>["http://files.figshare.com/1090990/Figure_4.tif"], "pos_in_sequence"=>0, "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "users"=>["Fabian Anderegg", "Marcus Textor", "Patrick Meraldi", "René Holtackers", "Mirren Charnley"], "links"=>[], "tags"=>["Cell physiology", "Extracellular matrix", "Biological systems engineering", "mitosis", "biotechnology", "spindle", "activation", "Musculoskeletal system", "cell adhesion", "cell division", "Bioengineering", "differences", "Anatomy and physiology", "biomechanics", "Cell mechanics", "Molecular cell biology", "checkpoint", "Extracellular matrix adhesions"], "title"=>"<p>The role of spindle checkpoint activation in the shape dependent differences in mitosis.</p>", "figshare_url"=>"http://figshare.com/articles/_The_role_of_spindle_checkpoint_activation_in_the_shape_dependent_differences_in_mitosis_/723547", "defined_type"=>1, "doi"=>["http://dx.doi.org/10.1371/journal.pone.0066918.g004"], "published_date"=>"2013-06-18 00:59:07", "article_id"=>723547, "categories"=>["Biological Sciences", "Engineering"], "description"=>"<p >HeLa (GFP-H2B/RFP-tubulin) cells were assessed for the parallel orientation of the mitotic spindle at metaphase after transfection with (A and B) scrambled siRNA or (C and D) Mad2 siRNA and culture in (A and C) square or (B and D) circular microwells to determine the role of spindle checkpoint activation. Cells cultured in circular microwells showed a random orientation of the mitotic spindle, whilst cells cultured in square microwells predominately aligned the spindle along the long axis of the cell, regardless of the depletion of Mad2. Similarly, the depletion of Mad2 had little effect on (E) the distribution and (F) average spindle orientation perpendicular to the substrate plane. Values represent spindle angle in degrees &#177; SEM. (G) Cells transfected with siRNA for Mad2 and cultured in circular microwells (white bars) took significantly less time to reach late prometaphase than cells transfected with control siRNA (dark grey bars), indicating that the shape dependent effect on the time required to reach late prometaphase was due to differences in spindle checkpoint activation. All values represent time in minutes &#177; SEM. Key: * p&lt;0.05, *** p&lt;0.001; NB &#8202;=&#8202; nuclear envelope breakdown, LPM &#8202;=&#8202; late prometaphase, M&#8202;=&#8202; metaphase, A&#8202;=&#8202; anaphase, C&#8202;=&#8202; cytokinesis.</p>"}
  • {"files"=>["http://files.figshare.com/1090991/Figure_5.tif"], "pos_in_sequence"=>0, "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "users"=>["Fabian Anderegg", "Marcus Textor", "Patrick Meraldi", "René Holtackers", "Mirren Charnley"], "links"=>[], "tags"=>["Extracellular matrix", "Biological systems engineering", "mitosis", "actin", "biotechnology", "differences", "Cell physiology", "Musculoskeletal system", "cell adhesion", "cell division", "Bioengineering", "Anatomy and physiology", "biomechanics", "cytoskeleton", "Cell mechanics", "Molecular cell biology", "Extracellular matrix adhesions"], "title"=>"<p>The role of the actin cytoskeleton in the shape dependent differences in mitosis.</p>", "figshare_url"=>"http://figshare.com/articles/_The_role_of_the_actin_cytoskeleton_in_the_shape_dependent_differences_in_mitosis_/723548", "defined_type"=>1, "doi"=>["http://dx.doi.org/10.1371/journal.pone.0066918.g005"], "published_date"=>"2013-06-18 00:59:08", "article_id"=>723548, "categories"=>["Biological Sciences", "Engineering"], "description"=>"<p >HeLa (YFP-paxillin) cells were cultured for 18 hours in either (A) square microwells or (B) circular microwells and imaged during interphase for actin (red) and paxillin (green); bars: 10 &#181;m. HeLa (GFP-H2B/RFP-tubulin) cells were assessed for the parallel orientation of the mitotic spindle at metaphase after culture in (C) square or (D) circular microwells and treatment with latrunculin A for 1 hour prior to mitosis. Cells cultured in circular microwells showed a random orientation of the mitotic spindle, whilst cells cultured in square microwells predominately aligned the spindle along the long axis of the cell. (E) The inhibition of actin polymerization in HeLa (GFP-H2B/RFP-tubulin) cells lead to increased tilting in cells cultured in the square microwells. Values represent spindle angle in degrees &#177; SEM. (F) Perturbation of actin polymerization in cells cultured in square microwells also increased the distribution of spindle orientation, but had little effect on cells cultured in circular microwells. (G) The perturbation of the actin cytoskeleton negated the differences in mitotic timings observed between cells cultured in square microwells (light grey bars) and cells cultured in circular microwells (dark grey bars). All values represent time in minutes &#177; SEM. NB &#8202;=&#8202; nuclear envelope breakdown, S&#8202;=&#8202; spindle formation, LPM &#8202;=&#8202; late prometaphase, M&#8202;=&#8202; metaphase, A&#8202;=&#8202; anaphase.</p>"}
  • {"files"=>["http://files.figshare.com/1090992/Figure_6.tif"], "pos_in_sequence"=>0, "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "users"=>["Fabian Anderegg", "Marcus Textor", "Patrick Meraldi", "René Holtackers", "Mirren Charnley"], "links"=>[], "tags"=>["2d", "Extracellular matrix", "Biological systems engineering", "patterned", "mitosis", "actin", "biotechnology", "Cell physiology", "Musculoskeletal system", "cell adhesion", "cell division", "Bioengineering", "Anatomy and physiology", "biomechanics", "cytoskeleton", "Cell mechanics", "Molecular cell biology", "Extracellular matrix adhesions"], "title"=>"<p>The effect of the actin cytoskeleton on mitosis after culture on 2D square patterned substrates.</p>", "figshare_url"=>"http://figshare.com/articles/_The_effect_of_the_actin_cytoskeleton_on_mitosis_after_culture_on_2D_square_patterned_substrates_/723549", "defined_type"=>1, "doi"=>["http://dx.doi.org/10.1371/journal.pone.0066918.g006"], "published_date"=>"2013-06-18 00:59:09", "article_id"=>723549, "categories"=>["Biological Sciences", "Engineering"], "description"=>"<p >(A) HeLa cells (YFP-paxillin) were cultured on 2D square patterns for 18 hours and imaged during interphase for actin (red) and paxillin (green). HeLa cells (RFP-tubulin/GFP-H2B) were assessed for the angle of the mitotic spindle in the xy plane at metaphase after culture on 2D square patterns and treatment with (B) media or (C) latrunculin A. A high proportion of the cells cultured on square patterns aligned the mitotic spindle along the diagonal of the cell, which was reduced by the addition of latrunculin A. (D) Conversely the addition of latrunculin A had little effect on the average orientation of the mitotic spindle perpendicular to the substrate plane. Values represent spindle angle in degrees &#177; SEM. (E) Furthermore, the addition of latrunculin A had little effect on the distribution of the spindle orientation. (F) Cells treated with latrunculin A (light grey bars) took significantly less time to progress from late prometaphase to metaphase and to complete mitosis than untreated cells (black bars). All values represent time in minutes &#177; SEM. Key: *** p&lt;0.001; NB &#8202;=&#8202; nuclear envelope breakdown, S&#8202;=&#8202; spindle formation, LPM &#8202;=&#8202; late prometaphase, M&#8202;=&#8202; metaphase, A&#8202;=&#8202; anaphase.</p>"}
  • {"files"=>["http://files.figshare.com/1090993/Table_1.xls"], "pos_in_sequence"=>0, "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "users"=>["Fabian Anderegg", "Marcus Textor", "Patrick Meraldi", "René Holtackers", "Mirren Charnley"], "links"=>[], "tags"=>["Extracellular matrix", "Biological systems engineering", "mitosis", "biotechnology", "Anatomy and physiology", "Cell physiology", "Musculoskeletal system", "cell adhesion", "cell division", "Bioengineering", "timings", "cultured", "biomechanics", "cells", "Cell mechanics", "Molecular cell biology", "mitotic", "Extracellular matrix adhesions"], "title"=>"<p>Analysis of mitotic timings in single cells cultured on different cell culture platforms.</p>", "figshare_url"=>"http://figshare.com/articles/_Analysis_of_mitotic_timings_in_single_cells_cultured_on_different_cell_culture_platforms_/723550", "defined_type"=>3, "doi"=>["http://dx.doi.org/10.1371/journal.pone.0066918.t001"], "published_date"=>"2013-06-18 00:59:10", "article_id"=>723550, "categories"=>["Biological Sciences", "Engineering"], "description"=>"<p>Analysis of mitotic timings in single cells cultured on different cell culture platforms.</p>"}
  • {"files"=>["http://files.figshare.com/1090994/Figure_S1.tif", "http://files.figshare.com/1090995/Figure_S2.tif", "http://files.figshare.com/1090996/Figure_S3.tif", "http://files.figshare.com/1090997/Video_S1.avi", "http://files.figshare.com/1090998/Video_S2.avi", "http://files.figshare.com/1090999/Video_S3.avi", "http://files.figshare.com/1091000/Video_S4.avi"], "pos_in_sequence"=>0, "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "users"=>["Fabian Anderegg", "Marcus Textor", "Patrick Meraldi", "René Holtackers", "Mirren Charnley"], "links"=>[], "tags"=>["Extracellular matrix", "Biological systems engineering", "mitosis", "biotechnology", "dimensionality", "spindle", "Cell physiology", "Musculoskeletal system", "cell adhesion", "cell division", "Bioengineering", "Anatomy and physiology", "biomechanics", "Cell mechanics", "Molecular cell biology", "mitotic", "Extracellular matrix adhesions"], "title"=>"<p>Effect of Cell Shape and Dimensionality on Spindle Orientation and Mitotic Timing</p>", "figshare_url"=>"http://figshare.com/articles/_Effect_of_Cell_Shape_and_Dimensionality_on_Spindle_Orientation_and_Mitotic_Timing_/723551", "defined_type"=>4, "doi"=>["http://dx.doi.org/10.1371/journal.pone.0066918.s001", "http://dx.doi.org/10.1371/journal.pone.0066918.s002", "http://dx.doi.org/10.1371/journal.pone.0066918.s003", "http://dx.doi.org/10.1371/journal.pone.0066918.s004", "http://dx.doi.org/10.1371/journal.pone.0066918.s005", "http://dx.doi.org/10.1371/journal.pone.0066918.s006", "http://dx.doi.org/10.1371/journal.pone.0066918.s007"], "published_date"=>"2013-06-18 00:59:11", "article_id"=>723551, "categories"=>["Biological Sciences", "Engineering"], "description"=>"<div><p>The formation and orientation of the mitotic spindle is a critical feature of mitosis. The morphology of the cell and the spatial distribution and composition of the cells' adhesive microenvironment all contribute to dictate the position of the spindle. However, the impact of the dimensionality of the cells' microenvironment has rarely been studied. In this study we present the use of a microwell platform, where the internal surfaces of the individual wells are coated with fibronectin, enabling the three-dimensional presentation of adhesive ligands to single cells cultured within the microwells. This platform was used to assess the effect of dimensionality and cell shape in a controlled microenvironment. Single HeLa cells cultured in circular microwells exhibited greater tilting of the mitotic spindle, in comparison to cells cultured in square microwells. This correlated with an increase in the time required to align the chromosomes at the metaphase plate due to prolonged activation of the spindle checkpoint in an actin dependent process. The comparison to 2D square patterns revealed that the dimensionality of cell adhesions alone affected both mitotic timings and spindle orientation; in particular the role of actin varied according to the dimensionality of the cells' microenvironment. Together, our data revealed that cell shape and the dimensionality of the cells' adhesive environment impacted on both the orientation of the mitotic spindle and progression through mitosis.</p></div>"}

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Relative Metric

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