Histamine reduces gap junctional communication of human tonsil high endothelial cells in culture

Xavier F. Figueroa, Karina Alviña, Agustín D. Martínez, Gladys Garcés, Mario Rosemblatt, Mauricio P. Boric, Juan C. Sáez

Resultado de la investigación: Article

12 Citas (Scopus)

Resumen

The regulation of gap junctional communication by histamine was studied in primary cultures of human tonsil high endothelial cells (HUTECs). We evaluated intercellular communication, levels, state of phosphorylation, and cellular distribution of gap junction protein subunits, mainly connexin (Cx)43. Histamine induced a time-dependent reduction in dye coupling (Lucifer yellow) associated with reduction in connexin43 localized at cell-cell appositions (immunofluorescence), without changes in levels and phosphorylation state of connexin43 (immunoblots). These effects were prevented with chlorpheniramine, an H1 receptor blocker; indomethacin, a cyclooxygenase blocker; or GF109203X, a protein kinase C inhibitor. Treatment with phorbol myristate acetate, a protein kinase C activator, and 4bromo (4Br)-A23187, a calcium ionophore, mimicked the histamine-induced effects on dye coupling. 8Bromo-cAMP doubled the dye coupling extent and prevented the histamine-induced reduction in incidence of dye coupling. After 24-h histamine treatment, known to desensitize H1 receptors, reapplication of histamine increased cell coupling in a way prevented by ranitidine, an H2 receptor blocker. Thus, activation of H1 and H2 receptors, which increase intracellular levels of free Ca2+ and cAMP, respectively, may affect gap junctional communication in opposite ways. Stabilization of actin filaments with phalloidine diminished but did not totally prevent histamine-induced cell shape changes and reduction in dye coupling. Hence, the histamine-induced reduction in gap junctional communication between HUTEC is mediated by cytoskeleton-dependent and -independent mechanisms and might contribute to modulate endothelial function in lymphoid tissue.

Idioma originalEnglish
Páginas (desde-hasta)247-257
Número de páginas11
PublicaciónMicrovascular Research
Volumen68
N.º3
DOI
EstadoPublished - 1 ene 2004

Huella dactilar

Palatine Tonsil
Endothelial cells
Cell culture
Histamine
Endothelial Cells
Cell Culture Techniques
Communication
Coloring Agents
Histamine H1 Receptors
Connexin 43
Histamine H2 Receptors
Phosphorylation
Protein Kinase C
Histamine Agents
Chlorpheniramine
Phalloidine
Connexins
Ranitidine
Calcium Ionophores
Cell Shape

ASJC Scopus subject areas

  • Biochemistry
  • Cardiology and Cardiovascular Medicine
  • Cell Biology

Citar esto

Figueroa, X. F., Alviña, K., Martínez, A. D., Garcés, G., Rosemblatt, M., Boric, M. P., & Sáez, J. C. (2004). Histamine reduces gap junctional communication of human tonsil high endothelial cells in culture. Microvascular Research, 68(3), 247-257. https://doi.org/10.1016/j.mvr.2004.06.009
Figueroa, Xavier F. ; Alviña, Karina ; Martínez, Agustín D. ; Garcés, Gladys ; Rosemblatt, Mario ; Boric, Mauricio P. ; Sáez, Juan C. / Histamine reduces gap junctional communication of human tonsil high endothelial cells in culture. En: Microvascular Research. 2004 ; Vol. 68, N.º 3. pp. 247-257.
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Figueroa, XF, Alviña, K, Martínez, AD, Garcés, G, Rosemblatt, M, Boric, MP & Sáez, JC 2004, 'Histamine reduces gap junctional communication of human tonsil high endothelial cells in culture', Microvascular Research, vol. 68, n.º 3, pp. 247-257. https://doi.org/10.1016/j.mvr.2004.06.009

Histamine reduces gap junctional communication of human tonsil high endothelial cells in culture. / Figueroa, Xavier F.; Alviña, Karina; Martínez, Agustín D.; Garcés, Gladys; Rosemblatt, Mario; Boric, Mauricio P.; Sáez, Juan C.

En: Microvascular Research, Vol. 68, N.º 3, 01.01.2004, p. 247-257.

Resultado de la investigación: Article

TY - JOUR

T1 - Histamine reduces gap junctional communication of human tonsil high endothelial cells in culture

AU - Figueroa, Xavier F.

AU - Alviña, Karina

AU - Martínez, Agustín D.

AU - Garcés, Gladys

AU - Rosemblatt, Mario

AU - Boric, Mauricio P.

AU - Sáez, Juan C.

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N2 - The regulation of gap junctional communication by histamine was studied in primary cultures of human tonsil high endothelial cells (HUTECs). We evaluated intercellular communication, levels, state of phosphorylation, and cellular distribution of gap junction protein subunits, mainly connexin (Cx)43. Histamine induced a time-dependent reduction in dye coupling (Lucifer yellow) associated with reduction in connexin43 localized at cell-cell appositions (immunofluorescence), without changes in levels and phosphorylation state of connexin43 (immunoblots). These effects were prevented with chlorpheniramine, an H1 receptor blocker; indomethacin, a cyclooxygenase blocker; or GF109203X, a protein kinase C inhibitor. Treatment with phorbol myristate acetate, a protein kinase C activator, and 4bromo (4Br)-A23187, a calcium ionophore, mimicked the histamine-induced effects on dye coupling. 8Bromo-cAMP doubled the dye coupling extent and prevented the histamine-induced reduction in incidence of dye coupling. After 24-h histamine treatment, known to desensitize H1 receptors, reapplication of histamine increased cell coupling in a way prevented by ranitidine, an H2 receptor blocker. Thus, activation of H1 and H2 receptors, which increase intracellular levels of free Ca2+ and cAMP, respectively, may affect gap junctional communication in opposite ways. Stabilization of actin filaments with phalloidine diminished but did not totally prevent histamine-induced cell shape changes and reduction in dye coupling. Hence, the histamine-induced reduction in gap junctional communication between HUTEC is mediated by cytoskeleton-dependent and -independent mechanisms and might contribute to modulate endothelial function in lymphoid tissue.

AB - The regulation of gap junctional communication by histamine was studied in primary cultures of human tonsil high endothelial cells (HUTECs). We evaluated intercellular communication, levels, state of phosphorylation, and cellular distribution of gap junction protein subunits, mainly connexin (Cx)43. Histamine induced a time-dependent reduction in dye coupling (Lucifer yellow) associated with reduction in connexin43 localized at cell-cell appositions (immunofluorescence), without changes in levels and phosphorylation state of connexin43 (immunoblots). These effects were prevented with chlorpheniramine, an H1 receptor blocker; indomethacin, a cyclooxygenase blocker; or GF109203X, a protein kinase C inhibitor. Treatment with phorbol myristate acetate, a protein kinase C activator, and 4bromo (4Br)-A23187, a calcium ionophore, mimicked the histamine-induced effects on dye coupling. 8Bromo-cAMP doubled the dye coupling extent and prevented the histamine-induced reduction in incidence of dye coupling. After 24-h histamine treatment, known to desensitize H1 receptors, reapplication of histamine increased cell coupling in a way prevented by ranitidine, an H2 receptor blocker. Thus, activation of H1 and H2 receptors, which increase intracellular levels of free Ca2+ and cAMP, respectively, may affect gap junctional communication in opposite ways. Stabilization of actin filaments with phalloidine diminished but did not totally prevent histamine-induced cell shape changes and reduction in dye coupling. Hence, the histamine-induced reduction in gap junctional communication between HUTEC is mediated by cytoskeleton-dependent and -independent mechanisms and might contribute to modulate endothelial function in lymphoid tissue.

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KW - Cytoskeleton

KW - Dye coupling

KW - Endothelium

KW - Gap junctions

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