Lagendijk Anne Karine, Gomez Guillermo A, Baek Sungmin, Hesselson Daniel, Hughes William E, Paterson Scott, Conway Daniel E, Belting Heinz-Georg, Affolter Markus, Smith Kelly A, Schwartz Martin A, Yap Alpha S, Hogan Benjamin M (2017), Live imaging molecular changes in junctional tension upon VE-cadherin in zebrafish., in Nature communications
, 8(1), 1402-1402.
Roubinet Chantal, Tsankova Anna, Pham Tri Thanh, Monnard Arnaud, Caussinus Emmanuel, Affolter Markus, Cabernard Clemens (2017), Spatio-temporally separated cortical flows and spindle geometry establish physical asymmetry in fly neural stem cells., in Nature communications
, 8(1), 1383-1383.
Ochoa-Espinosa Amanda, Harmansa Stefan, Caussinus Emmanuel, Affolter Markus (2017), Myosin II is not required for Drosophila tracheal branch elongation and cell intercalation., in Development (Cambridge, England)
, 144(16), 2961-2968.
Matsuda Shinya, Affolter Markus (2017), Dpp from the anterior stripe of cells is crucial for the growth of the Drosophila wing disc., in eLife
, 6, 1-13.
Sauteur Loïc, Affolter Markus, Belting Heinz-Georg (2017), Distinct and redundant functions of Esama and VE-cadherin during vascular morphogenesis., in Development (Cambridge, England)
, 144(8), 1554-1565.
Harmansa Stefan, Alborelli Ilaria, Bieli Dimitri, Caussinus Emmanuel, Affolter Markus (2017), A nanobody-based toolset to investigate the role of protein localization and dispersal in Drosophila., in eLife
, 6, 1-22.
Nakajima Hiroyuki, Yamamoto Kimiko, Agarwala Sobhika, Terai Kenta, Fukui Hajime, Fukuhara Shigetomo, Ando Koji, Miyazaki Takahiro, Yokota Yasuhiro, Schmelzer Etienne, Belting Heinz-Georg, Affolter Markus, Lecaudey Virginie, Mochizuki Naoki (2017), Flow-Dependent Endothelial YAP Regulation Contributes to Vessel Maintenance., in Developmental cell
, 40(6), 523-536.
Tsachaki Maria, Meyer Arne, Weger Benjamin, Kratschmar Denise V, Tokarz Janina, Adamski Jerzy, Belting Heinz-Georg, Affolter Markus, Dickmeis Thomas, Odermatt Alex (2017), Absence of 11-keto reduction of cortisone and 11-ketotestosterone in the model organism zebrafish., in The Journal of endocrinology
, 232(2), 323-335.
Caussinus Emmanuel, Affolter Markus (2016), deGradFP: A System to Knockdown GFP-Tagged Proteins., in Methods in molecular biology (Clifton, N.J.)
, 1478, 177-187.
Bieli D, Alborelli I, Harmansa S, Matsuda S, Caussinus E, Affolter M (2016), Development and Application of Functionalized Protein Binders in Multicellular Organisms., in International review of cell and molecular biology
, 325, 181-213.
Affolter Markus (2016), Seeing is Believing, or How GFP Changed My Approach to Science., in Current topics in developmental biology
, 116, 1-16.
Pasakarnis Laurynas, Frei Erich, Caussinus Emmanuel, Affolter Markus, Brunner Damian (2016), Amnioserosa cell constriction but not epidermal actin cable tension autonomously drives dorsal closure., in Nature cell biology
, 18(11), 1161-1172.
Betz Charles, Lenard Anna, Belting Heinz-Georg, Affolter Markus (2016), Cell behaviors and dynamics during angiogenesis., in Development (Cambridge, England)
, 143(13), 2249-60.
Bürglin Thomas R, Affolter Markus (2016), Homeodomain proteins: an update., in Chromosoma
, 125(3), 497-521.
Aydogan Vahap, Belting Heinz-Georg, Affolter Markus (2016), Mapping the molecular steps of secretory-lysosome-driven tracheal tube fusion., in Nature cell biology
, 18(7), 720-2.
Matsuda Shinya, Harmansa Stefan, Affolter Markus (2016), BMP morphogen gradients in flies., in Cytokine & growth factor reviews
, 27, 119-27.
Ittig Simon J, Schmutz Christoph, Kasper Christoph A, Amstutz Marlise, Schmidt Alexander, Sauteur Loïc, Vigano M Alessandra, Low Shyan Huey, Affolter Markus, Cornelis Guy R, Nigg Erich A, Arrieumerlou Cécile (2015), A bacterial type III secretion-based protein delivery tool for broad applications in cell biology., in The Journal of cell biology
, 211(4), 913-31.
Harmansa Stefan, Hamaratoglu Fisun, Affolter Markus, Caussinus Emmanuel (2015), Dpp spreading is required for medial but not for lateral wing disc growth., in Nature
, 527(7578), 317-22.
Bieli Dimitri, Kanca Oguz, Requena David, Hamaratoglu Fisun, Gohl Daryl, Schedl Paul, Affolter Markus, Slattery Matthew, Müller Martin, Estella Carlos (2015), Establishment of a Developmental Compartment Requires Interactions between Three Synergistic Cis-regulatory Modules., in PLoS genetics
, 11(10), 1005376-1005376.
Affolter Markus, Nigg Erich A (2015), In memoriam--Walter J. Gehring., in Chromosoma
, 124(3), 291-291.
Aydogan Vahap, Lenard Anna, Denes Alexandru Stefan, Sauteur Loic, Belting Heinz-Georg, Affolter Markus (2015), Endothelial cell division in angiogenic sprouts of differing cellular architecture., in Biology open
, 4(10), 1259-69.
Denes Alexandru Stefan, Kanca Oguz, Affolter Markus (2015), A cellular process that includes asymmetric cytokinesis remodels the dorsal tracheal branches in Drosophila larvae., in Development (Cambridge, England)
, 142(10), 1794-805.
Lenard Anna, Daetwyler Stephan, Betz Charles, Ellertsdottir Elin, Belting Heinz-Georg, Huisken Jan, Affolter Markus (2015), Endothelial cell self-fusion during vascular pruning., in PLoS biology
, 13(4), 1002126-1002126.
Bieli Dimitri, Kanca Oguz, Gohl Daryl, Denes Alexandru, Schedl Paul, Affolter Markus, Müller Martin (2015), The Drosophila melanogaster Mutants apblot and apXasta Affect an Essential apterous Wing Enhancer., in G3 (Bethesda, Md.)
, 5(6), 1129-43.
Phng Li-Kun, Gebala Véronique, Bentley Katie, Philippides Andrew, Wacker Andrin, Mathivet Thomas, Sauteur Loïc, Stanchi Fabio, Belting Heinz-Georg, Affolter Markus, Gerhardt Holger (2015), Formin-mediated actin polymerization at endothelial junctions is required for vessel lumen formation and stabilization., in Developmental cell
, 32(1), 123-32.
Brauchle Michael, Hansen Simon, Caussinus Emmanuel, Lenard Anna, Ochoa-Espinosa Amanda, Scholz Oliver, Sprecher Simon G, Plückthun Andreas, Affolter Markus (2014), Protein interference applications in cellular and developmental biology using DARPins that recognize GFP and mCherry., in Biology open
, 3(12), 1252-61.
Sauteur Loïc, Krudewig Alice, Herwig Lukas, Ehrenfeuchter Nikolaus, Lenard Anna, Affolter Markus, Belting Heinz-Georg (2014), Cdh5/VE-cadherin promotes endothelial cell interface elongation via cortical actin polymerization during angiogenic sprouting., in Cell reports
, 9(2), 504-13.
De Smet Frederik, Tembuyser Bieke, Lenard Anna, Claes Filip, Zhang Jie, Michielsen Christof, Van Schepdael Ann, Herbert Jean-Marc, Bono Françoise, Affolter Markus, Dewerchin Mieke, Carmeliet Peter (2014), Fibroblast growth factor signaling affects vascular outgrowth and is required for the maintenance of blood vessel integrity., in Chemistry & biology
, 21(10), 1310-1317.
Santos M Emília, Braasch Ingo, Boileau Nicolas, Meyer Britta S, Sauteur Loïc, Böhne Astrid, Belting Heinz-Georg, Affolter Markus, Salzburger Walter (2014), The evolution of cichlid fish egg-spots is linked with a cis-regulatory change., in Nature communications
, 5, 5149-5149.
Affolter Markus, Wüthrich Kurt (2014), Walter Jakob Gehring: A master of developmental biology., in Proceedings of the National Academy of Sciences of the United States of America
, 111(35), 12574-5.
Hamaratoglu Fisun, Affolter Markus, Pyrowolakis George (2014), Dpp/BMP signaling in flies: from molecules to biology., in Seminars in cell & developmental biology
, 32, 128-36.
Kanca Oguz, Ochoa-Espinosa Amanda, Affolter Markus (2014), IV. Tools and methods for studying cell migration and cell rearrangement in tissue and organ development., in Methods (San Diego, Calif.)
, 68(1), 228-32.
Hultin Sara, Zheng Yujuan, Mojallal Mahdi, Vertuani Simona, Gentili Christian, Balland Martial, Milloud Rachel, Belting Heinz-Georg, Affolter Markus, Helker Christian S M, Adams Ralf H, Herzog Wiebke, Uhlen Per, Majumdar Arindam, Holmgren Lars (2014), AmotL2 links VE-cadherin to contractile actin fibres necessary for aortic lumen expansion., in Nature communications
, 5, 3743-3743.
Kanca Oguz, Caussinus Emmanuel, Denes Alexandru S, Percival-Smith Anthony, Affolter Markus (2014), Raeppli: a whole-tissue labeling tool for live imaging of Drosophila development., in Development (Cambridge, England)
, 141(2), 472-80.
Project: In vivo cell biology of organ morphogenesis1 .Summary of the research planOrgans and tissues acquire particular three-dimensional shapes during development, which are intimately linked to the particular function(s) an organ has to fulfil. Organ shape is to a large extent determined by cell behaviour, and cell behaviour is to a large extent regulated by cell-cell signalling and cell mechanics. A major interest of my laboratory over the last few years has been to determine how branching morphogenesis restructures epithelial tissues to generate such fascinating structures as the trachea or the vasculature. While we have initially put much effort in understanding this process in vivo in an invertebrate system (tracheal development in Drosophila melanogaster) using state of the art genetics in combination with high resolution live imaging, we have recently moved much of our research efforts to vertebrates, in particular to zebrafish (Danio rerio). Using high resolution in vivo imaging, we have described the cellular activities during angiogenesis, in particular during sprouting, vessel fusion and vessel pruning. To our surprise, we find that the plasticity of developing vessels is accompanied by unexpected cell behaviour; endothelial cells can fission and self-fuse during anastomosis and pruning, respectively. With the advent of the recently introduced genome manipulation tools (TALEN, CRISPR/Cas9), a genetic dissection of the different steps in angiogenesis is now possible at unprecedented level, and proteins can be tagged at endogenous loci and used as marker for high resolution live imaging. We have recently applied intracellular nanobodies for the first time in developing drosophila embryos to directly manipulate protein function, in particular for protein degradation. We used an anti-GFP nanobody fused to an F-box in degrade GFP-fusion proteins (we called the method “deGradFP” for degrading GFP). In the meantime, we have functionalized the anti-GFP nanobody manifold, so that it can be used for in vivo protein trapping, protein localisation and for post-translation modification of proteins of interest. During the next granting period, we will introduce the use of intracellular nanobodies (and other protein binders) for studying the zebrafish vascular system, in particular to answer the following questions:Q1: How is the dynamic cell behaviour during sprouting controlled?Q2: How do endothelial cells recognize each other in order to connect?Q3: Which molecular processes are involved in luminal membrane expansion?Q4: Which molecular processes are involved in membrane fusion and membrane fission?In order to answer these questions, we propose to undertake the following experimental strategies:1) Investigate the role of VE-cad in the dynamic rearrangements of endothelial cells.2) Generate and use cutting edge in vivo live imaging tools to characterize molecular aspects of cell behaviour during angiogenesis processes.3) Generate and analyse mutations in candidate genes affecting distinct cellular activities.4) Generate and use novel tools to manipulate protein function in vivo - a step closer to a synthetic biology approach to organ formation.