Pflügers Archiv - European Journal of Physiology

, Volume 455, Issue 5, pp 859–872 | Cite as

Thrombin increases hyposmotic taurine efflux and accelerates \( {\text{ICI}}^{ - }_{{{\text{swell}}}} \) and RVD in 3T3 fibroblasts by a src-dependent EGFR transactivation

  • E. Vázquez-Juárez
  • G. Ramos-Mandujano
  • R. A. Lezama
  • S. Cruz-Rangel
  • L. D. Islas
  • H. Pasantes-MoralesEmail author
Cell and Molecular Physiology


The present study in Swiss3T3 fibroblasts examines the effect of thrombin on hyposmolarity-induced osmolyte fluxes and RVD, and the contribution of the src/EGFR pathway. Thrombin (5 U/ml) added to a 30% hyposmotic medium markedly increased hyposmotic 3H-taurine efflux (285%), accelerated the volume-sensitive Cl current (\( {\text{ICI}}^{ - }_{{{\text{swell}}}} \)) and increased RVD rate. These effects were reduced (50–65%) by preventing the thrombin-induced intracellular Ca2+ [Ca2+]i rise with EGTA-AM, or with the phospholipase C (PLC) blocker U73122. Ca2+calmodulin (CaM) and calmodulin kinase II (CaMKII) also participate in this Ca2+-dependent pathway. Thrombin plus hyposmolarity increased src and EGFR phosphorylation, whose blockade by PP2 and AG1478, decreased by 30–50%, respectively, the thrombin effects on hyposmotic taurine efflux, \( {\text{ICI}}^{ - }_{{{\text{swell}}}} \) and RVD. Ca2+- and src/EGFR-mediated pathways operate independently as shown by (1) the persistence of src and EGFR activation when [Ca2+]i rise is prevented and (2) the additive effect on taurine efflux, \( {\text{ICI}}^{ - }_{{{\text{swell}}}} \) or RVD by simultaneous inhibition of the two pathways, which essentially suppressed these events. PLC–Ca2+- and src/EGFR-signaling pathways operate in the hyposmotic condition and because thrombin per se failed to increase taurine efflux and \( {\text{ICI}}^{ - }_{{{\text{swell}}}} \) under isosmotic condition it seems that it is merely amplifying these previously activated mechanisms. The study shows that thrombin potentiates hyposmolarity-induced osmolyte fluxes and RVD by increasing src/EGFR-dependent signaling, in addition to the Ca2+-dependent pathway.


Thrombin Volume regulation Volume Taurine Swelling-activated chloride channel Swelling 



We deeply appreciate the technical assistance of Ms. Claudia Peña Segura. This study was supported in part by grant nos. IN209507 from DGAPA-UNAM and 46465 from CONACYT.


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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • E. Vázquez-Juárez
    • 1
  • G. Ramos-Mandujano
    • 1
  • R. A. Lezama
    • 3
  • S. Cruz-Rangel
    • 1
  • L. D. Islas
    • 2
  • H. Pasantes-Morales
    • 1
    • 4
    Email author
  1. 1.Departamento de Biofísica, Instituto de Fisiología CelularUniversidad Nacional Autónoma de MéxicoMéxicoMéxico
  2. 2.Departamento de Fisiología, Facultad de MedicinaUniversidad Nacional Autónoma de MéxicoMéxicoMéxico
  3. 3.Escuela Nacional de Ciencias BiológicasInstituto Politécnico NacionalMéxicoMéxico
  4. 4.Instituto de Fisiología CelularUNAMMéxicoMéxico

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