title: Claudin-5 controls intercellular barriers of human dermal microvascular but not human umbilical vein endothelial cells.
abstract: OBJECTIVE
  To assess the role claudin-5, an endothelial cell (EC) tight junction protein, plays in establishing basal permeability levels in humans by comparing claudin-5 expression levels in situ and analyzing junctional organization and function in 2 widely used models of cultured ECs, namely human dermal microvascular (HDM)ECs and human umbilical vein (HUV)ECs.
  
  
  RESULTS
  By immunofluorescence microscopy, ECs more highly express claudin-5 (but equivalently express vascular endothelial-cadherin) in human dermal capillaries versus postcapillary venules and in umbilical and coronary arteries versus veins, correlating with known segmental differences in tight junction frequencies and permeability barriers. Postconfluent cultured HDMECs express more claudin-5 (but equivalent vascular endothelial-cadherin) and show higher transendothelial electric resistance and lower macromolecular flux than similarly cultured HUVECs. HDMEC junctions are more complex by transmission electron microscopy and show more continuous claudin-5 immunofluorescence than HUVEC junctions. Calcium chelation or dominant negative vascular endothelial-cadherin overexpression decreases transendothelial electric resistance and disrupts junctions in HUVECs, but not in HDMECs. Claudin-5 overexpression in HUVECs fails to increase transendothelial electric resistance or claudin-5 continuity, whereas claudin-5 knockdown in HDMECs, but not in HUVECs, reduces transendothelial electric resistance and increases antibody accessibility to junctional proteins.
  
  
  CONCLUSIONS
  Claudin-5 expression and junctional organization control HDMEC and arteriolar-capillary paracellular barriers, whereas HUVEC and venular junctions use vascular endothelial-cadherin.
authors.affiliation: Department of Immunobiology, Yale University School of Medicine, New Haven, CT 06519, USA. martin.kluger@yale.edu
authors.forename: Martin S
authors.initials: MS
authors.lastname: Kluger
authors.forename: Paul R
authors.initials: PR
authors.lastname: Clark
authors.forename: George
authors.initials: G
authors.lastname: Tellides
authors.forename: Volker
authors.initials: V
authors.lastname: Gerke
authors.forename: Jordan S
authors.initials: JS
authors.lastname: Pober
mesh_terms: D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002199:Capillary Permeability / Q000187:drug effects; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D062507:Claudin-5 / Q000235:genetics / Q000378:metabolism*; D017097:Electric Impedance; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D055785:Gene Knockdown Techniques; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D055806:Microvessels / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D012867:Skin / Q000098:blood supply*; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D013997:Time Factors; D014162:Transfection; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D002118:Calcium; D002614:Chelating Agents; D062507:Claudin-5; D000099152:Cadherin 5; C107466:CLDN5 protein, human