Cell Substratum Adhesion during Early Development of Dictyostelium discoideum
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{"title"=>"Cell substratum adhesion during early development of dictyostelium discoideum", "type"=>"journal", "authors"=>[{"first_name"=>"Marco", "last_name"=>"Tarantola", "scopus_author_id"=>"25723913400"}, {"first_name"=>"Albert", "last_name"=>"Bae", "scopus_author_id"=>"6602324786"}, {"first_name"=>"Danny", "last_name"=>"Fuller", "scopus_author_id"=>"7102775433"}, {"first_name"=>"Eberhard", "last_name"=>"Bodenschatz", "scopus_author_id"=>"7004288217"}, {"first_name"=>"Wouter Jan", "last_name"=>"Rappel", "scopus_author_id"=>"6603867039"}, {"first_name"=>"William F.", "last_name"=>"Loomis", "scopus_author_id"=>"35370097300"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203", "scopus"=>"2-s2.0-84907478636", "pui"=>"600067509", "doi"=>"10.1371/journal.pone.0106574", "issn"=>"19326203", "pmid"=>"25247557", "sgr"=>"84907478636"}, "id"=>"b1842cbd-1ed9-3840-aa19-4eef5ca6c7d3", "abstract"=>"Vegetative and developed amoebae of Dictyostelium discoideum gain traction and move rapidly on a wide range of substrata without forming focal adhesions. We used two independent assays to quantify cell-substrate adhesion in mutants and in wild-type cells as a function of development. Using a microfluidic device that generates a range of hydrodynamic shear stress, we found that substratum adhesion decreases at least 10 fold during the first 6 hr of development of wild type cells. This result was confirmed using a single-cell assay in which cells were attached to the cantilever of an atomic force probe and allowed to adhere to untreated glass surfaces before being retracted. Both of these assays showed that the decrease in substratum adhesion was dependent on the cAMP receptor CAR1 which triggers development. Vegetative cells missing talin as the result of a mutation in talA exhibited slightly reduced adhesive properties compared to vegetative wild-type cells. In sharp contrast to wild-type cells, however, these talA mutant cells did not show further reduction of adhesion during development such that after 5 hr of development they were significantly more adhesive than developed wild type cells. In addition, both assays showed that substrate adhesion was reduced in 0 hr cells when the actin cytoskeleton was disrupted by latrunculin. Consistent with previous observations, substrate adhesion was also reduced in 0 hr cells lacking the membrane proteins SadA or SibA as the result of mutations in sadA or sibA. However, there was no difference in the adhesion properties between wild type AX3 cells and these mutant cells after 6 hr of development, suggesting that neither SibA nor SadA play an essential role in substratum adhesion during aggregation. Our results provide a quantitative framework for further studies of cell substratum adhesion in Dictyostelium.", "link"=>"http://www.mendeley.com/research/cell-substratum-adhesion-during-early-development-dictyostelium-discoideum", "reader_count"=>18, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>4, "Researcher"=>5, "Student > Ph. D. Student"=>1, "Student > Master"=>4, "Student > Bachelor"=>2, "Student > Doctoral Student"=>2}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>4, "Researcher"=>5, "Student > Ph. D. Student"=>1, "Student > Master"=>4, "Student > Bachelor"=>2, "Student > Doctoral Student"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>2, "Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>6, "Physics and Astronomy"=>5}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>2}, "Physics and Astronomy"=>{"Physics and Astronomy"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>6}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1686593"], "description"=>"<p><b>A</b> Wild type (AX3) and <i>carA</i><sup>−</sup> cells lacking the cAMP receptor after 0 and 5 hr of development were analyzed with the microfluidic assay for the remaining fraction of cells after 40 minutes. <b>B</b> Work of adhesion (W<sub>Adh</sub>.) of wild type (AX3), cells lacking the cAMP receptor (<i>carA</i><sup>−</sup>) or cells lacking talin (<i>talA<sup>−</sup></i>) was measured after 0 hr (veg <i>n</i> = 36) and 6 hr (dev <i>n</i> = 30 for <i>carA<sup>−</sup></i>, <i>n</i> = 34 for <i>talA<sup>−</sup></i>) of development. Nonparametric statistical hypothesis test Wilcoxon rank-sum test was used for significance.</p>", "links"=>[], "tags"=>["0 hr cells", "type cells", "cell Substratum Adhesion", "substratum adhesion", "type AX 3 cells", "Dictyostelium discoideum gain traction", "6 hr", "substrate adhesion", "substratum adhesion decreases", "hydrodynamic shear stress", "Dictyostelium discoideum Vegetative", "cAMP receptor CAR 1", "membrane proteins SadA"], "article_id"=>1178632, "categories"=>["Biological Sciences"], "users"=>["Marco Tarantola", "Albert Bae", "Danny Fuller", "Eberhard Bodenschatz", "Wouter-Jan Rappel", "William F. Loomis"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106574.g003", "stats"=>{"downloads"=>0, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Developmental_regulation_of_the_decrease_in_adhesion_/1178632", "title"=>"Developmental regulation of the decrease in adhesion.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-23 03:02:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/1686591"], "description"=>"<p><b>A.</b> Force was measured from bending of the cantilever and a typical force-separation curve is shown for an approach-retraction cycle highlighting the two assessed parameters: maximum adhesion force F<sub>Max.Adh.</sub> and the work of adhesion W<sub>Adh.</sub> (integral of the hatched area). I–IV refers to the four panels shown under B. <b>B.</b> Side view of a cycle of approach and retraction of a cell attached to a cantilever. Panels I and II: A cell can be seen hanging below the cantilever as it approaches the glass slide (light colored surface, <i>Dictyostelium</i> contour in red under I). Panels III and IV: The cell can be seen to remain on the cantilever as it is retracted and it maintains a rounded shape. Arrows highlight cell position. Scale bar is 20 µm.</p>", "links"=>[], "tags"=>["0 hr cells", "type cells", "cell Substratum Adhesion", "substratum adhesion", "type AX 3 cells", "Dictyostelium discoideum gain traction", "6 hr", "substrate adhesion", "substratum adhesion decreases", "hydrodynamic shear stress", "Dictyostelium discoideum Vegetative", "cAMP receptor CAR 1", "membrane proteins SadA"], "article_id"=>1178630, "categories"=>["Biological Sciences"], "users"=>["Marco Tarantola", "Albert Bae", "Danny Fuller", "Eberhard Bodenschatz", "Wouter-Jan Rappel", "William F. Loomis"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106574.g001", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Single_Cell_adhesion_Force_Spectroscopy_assay_/1178630", "title"=>"Single Cell adhesion Force Spectroscopy assay.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-23 03:02:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/1686594"], "description"=>"<p><b>A</b> Cell-substratum adhesion of wild type Ax3, <i>sibA</i> and <i>sadA</i> null cells at either 0 or 5 hr of development as measured by the microfluidic assay from the remaining fraction of cells after 40 minutes. <b>B</b> Work of adhesion W<sub>Adh.</sub> of wild type (AX3) and cells lacking either <i>sibA</i> or <i>sadA</i> was measured after 0 hr (<i>n</i> = 29 and 41) and 6 hr (<i>n</i> = 45 and 41) of development. Significance was judged from the Wilcoxon-rank-sum test.</p>", "links"=>[], "tags"=>["0 hr cells", "type cells", "cell Substratum Adhesion", "substratum adhesion", "type AX 3 cells", "Dictyostelium discoideum gain traction", "6 hr", "substrate adhesion", "substratum adhesion decreases", "hydrodynamic shear stress", "Dictyostelium discoideum Vegetative", "cAMP receptor CAR 1", "membrane proteins SadA"], "article_id"=>1178633, "categories"=>["Biological Sciences"], "users"=>["Marco Tarantola", "Albert Bae", "Danny Fuller", "Eberhard Bodenschatz", "Wouter-Jan Rappel", "William F. Loomis"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106574.g004", "stats"=>{"downloads"=>0, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Substratum_adhesion_in_cells_lacking_sibA_or_s_adA_/1178633", "title"=>"Substratum adhesion in cells lacking <i>sibA</i> or s<i>adA</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-23 03:02:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/1686595", "https://ndownloader.figshare.com/files/1686596", "https://ndownloader.figshare.com/files/1686597", "https://ndownloader.figshare.com/files/1686598", "https://ndownloader.figshare.com/files/1686599", "https://ndownloader.figshare.com/files/1686600"], "description"=>"<div><p>Vegetative and developed amoebae of <i>Dictyostelium discoideum</i> gain traction and move rapidly on a wide range of substrata without forming focal adhesions. We used two independent assays to quantify cell-substrate adhesion in mutants and in wild-type cells as a function of development. Using a microfluidic device that generates a range of hydrodynamic shear stress, we found that substratum adhesion decreases at least 10 fold during the first 6 hr of development of wild type cells. This result was confirmed using a single-cell assay in which cells were attached to the cantilever of an atomic force probe and allowed to adhere to untreated glass surfaces before being retracted. Both of these assays showed that the decrease in substratum adhesion was dependent on the cAMP receptor CAR1 which triggers development. Vegetative cells missing talin as the result of a mutation in <i>talA</i> exhibited slightly reduced adhesive properties compared to vegetative wild-type cells. In sharp contrast to wild-type cells, however, these <i>talA</i> mutant cells did not show further reduction of adhesion during development such that after 5 hr of development they were significantly more adhesive than developed wild type cells. In addition, both assays showed that substrate adhesion was reduced in 0 hr cells when the actin cytoskeleton was disrupted by latrunculin. Consistent with previous observations, substrate adhesion was also reduced in 0 hr cells lacking the membrane proteins SadA or SibA as the result of mutations in <i>sadA</i> or <i>sibA</i>. However, there was no difference in the adhesion properties between wild type AX3 cells and these mutant cells after 6 hr of development, suggesting that neither SibA nor SadA play an essential role in substratum adhesion during aggregation. Our results provide a quantitative framework for further studies of cell substratum adhesion in <i>Dictyostelium</i>.</p></div>", "links"=>[], "tags"=>["0 hr cells", "type cells", "cell Substratum Adhesion", "substratum adhesion", "type AX 3 cells", "Dictyostelium discoideum gain traction", "6 hr", "substrate adhesion", "substratum adhesion decreases", "hydrodynamic shear stress", "Dictyostelium discoideum Vegetative", "cAMP receptor CAR 1", "membrane proteins SadA"], "article_id"=>1178634, "categories"=>["Biological Sciences"], "users"=>["Marco Tarantola", "Albert Bae", "Danny Fuller", "Eberhard Bodenschatz", "Wouter-Jan Rappel", "William F. Loomis"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0106574.s001", "https://dx.doi.org/10.1371/journal.pone.0106574.s002", "https://dx.doi.org/10.1371/journal.pone.0106574.s003", "https://dx.doi.org/10.1371/journal.pone.0106574.s004", "https://dx.doi.org/10.1371/journal.pone.0106574.s005", "https://dx.doi.org/10.1371/journal.pone.0106574.s006"], "stats"=>{"downloads"=>1, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Cell_Substratum_Adhesion_during_Early_Development_of_Dictyostelium_discoideum_/1178634", "title"=>"Cell Substratum Adhesion during Early Development of <i>Dictyostelium discoideum</i>", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-09-23 03:02:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/1686592"], "description"=>"<p><b>A.</b> Microfluidic assay: the remaining fraction of cells after 40 minutes in chambers 2–8 is shown for cells that had developed for varying lengths of time. latB refers to 0 hr cells that were treated with 10 µM latrunculin B for 30 minutes to disrupt the actin cytoskeleton and assayed in the presence of the drug. Average of at least 5 independent experiments. The bars indicate the S.E.M. <b>B.</b> Single cell adhesion force assay of cells during early development. Box plot of the distribution of maximum adhesion forces F<sub>Max.Adh</sub> of individual single cells, where the bottom and the top of the box represents the first and the third quartiles, and the band corresponds to the median. Cells were developed for 0 h (<i>n</i> = 33), 3 h (<i>n</i> = 47), 6 hr (<i>n</i> = 31). Top whiskers are at 90% and bottom whiskers are at 10% of the distribution. Latrunculin B treated 0 hr cells (Lat) (<i>n</i> = 27) were assayed in the presence of 10 µM latrunculin B to disrupt the actin cytoskeleton. <b>C.</b> Work of adhesion W<sub>Adh.</sub> measured for the same cells. Nonparametric statistical hypothesis test Wilcoxon rank-sum test were used for significance.</p>", "links"=>[], "tags"=>["0 hr cells", "type cells", "cell Substratum Adhesion", "substratum adhesion", "type AX 3 cells", "Dictyostelium discoideum gain traction", "6 hr", "substrate adhesion", "substratum adhesion decreases", "hydrodynamic shear stress", "Dictyostelium discoideum Vegetative", "cAMP receptor CAR 1", "membrane proteins SadA"], "article_id"=>1178631, "categories"=>["Biological Sciences"], "users"=>["Marco Tarantola", "Albert Bae", "Danny Fuller", "Eberhard Bodenschatz", "Wouter-Jan Rappel", "William F. Loomis"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106574.g002", "stats"=>{"downloads"=>1, "page_views"=>22, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Decrease_in_adhesion_during_early_development_of_Dictyostelium_discoideum_/1178631", "title"=>"Decrease in adhesion during early development of <i>Dictyostelium discoideum</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-23 03:02:11"}

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