Effect of cyclic AMP on barrier function of human lymphatic microvascular tubes.

This work examines the effect of cyclic AMP (cAMP) on the in vitro barrier function of tubes of human dermal lymphatic microvascular endothelial cells (LECs). Under baseline conditions, the barrier function of LEC tubes was weak, with diffusional permeability coefficients to bovine serum albumin and 10 kDa dextran of 1.4(-0.6)(+0.9)x10(-6) cm/s and 1.7(-0.5)(+0.8)x10(-6) cm/s (geometric mean+/-95% CI), respectively, and 1.2+/-0.5 (mean+/-95% CI) focal leaks per mm. Exposure to low concentrations (3 microM) of a cell-permeant analog of cAMP did not alter the barrier function. Exposure to higher concentrations (80 and 400 microM) and/or the phosphodiesterase inhibitor Ro-20-1724 (20 microM) lowered permeabilities and the number of focal leaks, and increased the selectivity of the barrier. Decreased permeabilities were accompanied by an increase in continuous VE-cadherin staining at cell-cell borders. Exposure to 1 mM 2',5'-dideoxyadenosine, an inhibitor of adenylate cyclase, did not increase permeabilities. LECs expressed the lymphatic-specific master transcription factor Prox-1, regardless of whether barrier function was weak or strong. Our results indicate that the permeability of LEC tubes in vitro responds to cAMP in a manner similar to that well-described for the permeability of blood microvessels.

[1]  J. Cooper,et al.  Endothelial barrier function. , 1989, The Journal of investigative dermatology.

[2]  L. Rubin,et al.  Erratum: Microvascular permeability and number of tight junctions are modulated by cAMP (American Journal of Physiology Heart an Circulatory Physiology (1998) 274:43 (H1885-H1894)) , 1998 .

[3]  F. Curry,et al.  Dominant role of cAMP in regulation of microvessel permeability. , 2000, American journal of physiology. Heart and circulatory physiology.

[4]  H. Dvorak,et al.  Pathways of macromolecular tracer transport across venules and small veins. Structural basis for the hyperpermeability of tumor blood vessels. , 1992, Laboratory investigation; a journal of technical methods and pathology.

[5]  S. Holgate,et al.  Role of adenylate cyclase in immunologic release of mediators from rat mast cells: agonist and antagonist effects of purine- and ribose-modified adenosine analogs. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[6]  M. Lampugnani,et al.  Interendothelial junctions: structure, signalling and functional roles. , 1997, Current opinion in cell biology.

[7]  T. Libermann,et al.  Identification of vascular lineage-specific genes by transcriptional profiling of isolated blood vascular and lymphatic endothelial cells. , 2003, The American journal of pathology.

[8]  M. Detmar,et al.  An essential role for Prox1 in the induction of the lymphatic endothelial cell phenotype , 2002, The EMBO journal.

[9]  A. Bollinger,et al.  Permeability of superficial lymphatic capillaries in human skin to FITC-labelled dextrans 40 000 and 150 000. , 1984, International journal of microcirculation, clinical and experimental.

[10]  S. Dudek,et al.  Cytoskeletal regulation of pulmonary vascular permeability. , 2001, Journal of applied physiology.

[11]  J. Burke,et al.  Fine structure of the lymphatic capillary and the adjoining connective tissue area. , 1966, The American journal of anatomy.

[12]  J. Cooper,et al.  Measurement of albumin permeability across endothelial monolayers in vitro. , 1987, Journal of applied physiology.

[13]  B. Zweifach,et al.  Micromanipulation of pressure in terminal lymphatics in the mesentery. , 1975, The American journal of physiology.

[14]  F. Rost.  Quantitative fluorescence microscopy , 1991 .

[15]  W. Risau,et al.  Modulation of tight junction structure in blood-brain barrier endothelial cells. Effects of tissue culture, second messengers and cocultured astrocytes. , 1994, Journal of cell science.

[16]  M. Skobe,et al.  Molecular characterization of lymphatic endothelial cells , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[17]  G. Palade,et al.  Transcytosis of albumin in capillary endothelium , 1987, The Journal of cell biology.

[18]  E. M. Renkin,et al.  Transport of large molecules from plasma to interstitial fluid and lymph in dogs. , 1970, The American journal of physiology.

[19]  K. Alitalo,et al.  Lymphatic endothelial reprogramming of vascular endothelial cells by the Prox‐1 homeobox transcription factor , 2002, The EMBO journal.

[20]  G. Schmid-Schönbein,et al.  Microlymphatics and lymph flow. , 1990, Physiological reviews.

[21]  J. Dye,et al.  Cyclic AMP and acidic fibroblast growth factor have opposing effects on tight and adherens junctions in microvascular endothelial cells in vitro. , 2001, Microvascular research.

[22]  A. Verin,et al.  Sphingosine 1-phosphate promotes endothelial cell barrier integrity by Edg-dependent cytoskeletal rearrangement. , 2001, The Journal of clinical investigation.

[23]  F. Curry,et al.  Microvascular permeability. , 1999, Physiological reviews.

[24]  R. Schaeffer,et al.  Effects of human alpha-thrombin and 8bromo-cAMP on large and microvessel endothelial monolayer equivalent "pore" radii. , 1995, Microvascular Research.

[25]  L. Leak.  STUDIES ON THE PERMEABILITY OF LYMPHATIC CAPILLARIES , 1971, The Journal of cell biology.

[26]  A. Taylor,et al.  Reversal of pulmonary capillary ischemia-reperfusion injury by rolipram, a cAMP phosphodiesterase inhibitor. , 1994, Journal of applied physiology.

[27]  Joe Tien,et al.  Formation of perfused, functional microvascular tubes in vitro. , 2006, Microvascular research.

[28]  Dick W. Slaaf,et al.  The endothelial glycocalyx: composition, functions, and visualization , 2007, Pflügers Archiv - European Journal of Physiology.

[29]  H. Barth,et al.  cAMP protects endothelial barrier functions by preventing Rac-1 inhibition. , 2004, American journal of physiology. Heart and circulatory physiology.

[30]  Shoichiro Tsukita,et al.  Multifunctional strands in tight junctions , 2001, Nature Reviews Molecular Cell Biology.

[31]  C. C. O'morchoe,et al.  An ultrastructural study of transport pathways across rat hepatic lymph vessels. , 1985, Lymphology.

[32]  G. Sacchi,et al.  Plasticity of Intercellular Junctions of Rat Liver Lymph Capillaries in Relation to Functional Conditions , 1991, Angiology.

[33]  J. Katz,et al.  Method for determination of cyclic AMP in plasma. , 1973, Clinical chemistry.

[34]  B. Geiger,et al.  The molecular organization of endothelial cell to cell junctions: differential association of plakoglobin, beta-catenin, and alpha- catenin with vascular endothelial cadherin (VE-cadherin) , 1995, The Journal of cell biology.

[35]  R. Jain,et al.  Microvascular permeability of normal and neoplastic tissues. , 1986, Microvascular research.

[36]  S. Amatschek,et al.  Blood and lymphatic endothelial cell-specific differentiation programs are stringently controlled by the tissue environment. , 2007, Blood.

[37]  L. Rubin,et al.  A cell culture model of the blood-brain barrier , 1991, The Journal of cell biology.

[38]  M. Rode,et al.  Induction of endothelial barrier function in vitro. , 2006, Annals of the New York Academy of Sciences.

[39]  D. Kerjaschki,et al.  Isolation and Characterization of Dermal Lymphatic and Blood Endothelial Cells Reveal Stable and Functionally Specialized Cell Lineages , 2001, The Journal of experimental medicine.

[40]  Thomas N. Sato,et al.  Leakage-resistant blood vessels in mice transgenically overexpressing angiopoietin-1. , 1999, Science.

[41]  M. Farquhar,et al.  THE PERMEABILITY OF GLOMERULAR CAPILLARIES TO GRADED DEXTRANS , 1974, The Journal of cell biology.

[42]  S. Butz,et al.  Functionally specialized junctions between endothelial cells of lymphatic vessels , 2007 .

[43]  H. Davis,et al.  Mechanisms of cholera toxin prevention of thrombin- and PMA-induced endothelial cell barrier dysfunction. , 1994, Microvascular research.

[44]  L. Rubin,et al.  Microvascular permeability and number of tight junctions are modulated by cAMP. , 1998, American journal of physiology. Heart and circulatory physiology.

[45]  W. K. Adkins,et al.  Compounds that increase cAMP prevent ischemia-reperfusion pulmonary capillary injury. , 1992, Journal of applied physiology.

[46]  J. Siliciano,et al.  Identification of ZO-1: a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia , 1986, The Journal of cell biology.

[47]  A. Fishman,et al.  Permeability characteristics of cultured endothelial cell monolayers. , 1988, Journal of applied physiology.

[48]  A. Taylor,et al.  Reversal of increased microvascular permeability associated with ischemia-reperfusion: role of cAMP. , 1992, Journal of applied physiology.

[49]  M. Detmar,et al.  Up-regulation of the lymphatic marker podoplanin, a mucin-type transmembrane glycoprotein, in human squamous cell carcinomas and germ cell tumors. , 2005, The American journal of pathology.

[50]  T. Ohhashi,et al.  Effective permeability of hydrophilic substances through walls of lymph vessels: roles of endothelial barrier. , 2005, American journal of physiology. Heart and circulatory physiology.

[51]  L. McIntire,et al.  Permeability of human endothelial monolayers: effect of vasoactive agonists and cAMP. , 1989, Journal of applied physiology.