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@ARTICLE{LeBihan:141262,
      author       = {Le Bihan, Olivier and Decossas, Marion and Gontier, Etienne
                      and Gerbod-Giannone, Marie-Christine and Lambert, Olivier},
      title        = {{V}isualization of adherent cell monolayers by
                      cryo-electron microscopy: {A} snapshot of endothelial
                      adherens junctions.},
      journal      = {Journal of structural biology},
      volume       = {192},
      number       = {3},
      issn         = {1047-8477},
      address      = {San Diego, Calif.},
      publisher    = {Elsevier},
      reportid     = {DZNE-2020-07584},
      pages        = {470-477},
      year         = {2015},
      abstract     = {Cryo-electron microscopy (cryo-EM) allows the visualization
                      of the cell architecture in its native state. We developed a
                      robust solution to adapt cryo-electron microscopy of
                      vitreous sections (CEMOVIS) to a monolayer of adherent cells
                      using a functionalized polyacrylamide hydrogel growing
                      substrate. We applied this method to reconstitute an
                      endothelial cell monolayer to visualize the morphology of
                      adherens junctions (AJs) which regulate permeability and
                      integrity of the vascular barrier. The fine morphology and
                      ultrastructure of AJs from cultured primary human coronary
                      artery endothelial cells (HCAECs) were analyzed in their
                      native state by using CEMOVIS. Doxycycline and
                      sphingosine-1-phosphate (S1P) are known as efficient
                      regulators of endothelial permeability. Doxycycline and S1P
                      treatments both led to a drastic morphological switch from
                      very uneven to standardized 14-17 nm wide AJs over several
                      microns indicative of a better membrane tethering.
                      Repetitive structures were occasionally noticed within the
                      AJ cleft reflecting a local improved structural organization
                      of VE-cadherin molecules. The ultrastructural stabilization
                      of AJs observed upon treatment likely indicates a better
                      adhesion and thus provides structural clues on the mechanism
                      by which these treatments improve the endothelial barrier
                      function. This method was also successfully extended to a
                      thick epithelial barrier model. We expect our strategy to
                      extend the reliable application of CEMOVIS to virtually any
                      adherent cultured cell systems.},
      keywords     = {Adherens Junctions: physiology / Antigens, CD: metabolism /
                      Caco-2 Cells / Cadherins: metabolism / Cell Line, Tumor /
                      Coronary Vessels: cytology / Cryoelectron Microscopy:
                      methods / Doxycycline: pharmacology / Endothelial Cells:
                      cytology / Humans / Lysophospholipids: pharmacology /
                      Sphingosine: analogs $\&$ derivatives / Sphingosine:
                      pharmacology / Staining and Labeling / Antigens, CD (NLM
                      Chemicals) / Cadherins (NLM Chemicals) / Lysophospholipids
                      (NLM Chemicals) / cadherin 5 (NLM Chemicals) / sphingosine
                      1-phosphate (NLM Chemicals) / Doxycycline (NLM Chemicals) /
                      Sphingosine (NLM Chemicals)},
      cin          = {AG Alamoudi},
      ddc          = {540},
      cid          = {I:(DE-2719)1013012},
      pnm          = {341 - Molecular Signaling (POF3-341)},
      pid          = {G:(DE-HGF)POF3-341},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:26470813},
      doi          = {10.1016/j.jsb.2015.10.009},
      url          = {https://pub.dzne.de/record/141262},
}