Reorganization of ZO-1 by sodium-dependent glucose transporter activation after heat stress in LLC-PK1 cells
| Autor: | Yasunobu Suketa, Mika Nakano, Akira Ikari, Kuniaki Takagi, Hitoshi Harada |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Cell Membrane Permeability
Monosaccharide Transport Proteins Swine Physiology Clinical Biochemistry Genistein Cell Communication Membrane Potentials Tight Junctions Diffusion chemistry.chemical_compound Cytosol Sodium-Glucose Transporter 1 Electric Impedance Animals Enzyme Inhibitors Incubation Membrane Glycoproteins Tight junction Glucose transporter Membrane Proteins Dextrans Epithelial Cells Glucose analog Recovery of Function Cell Biology Protein-Tyrosine Kinases Phosphoproteins Molecular biology Up-Regulation Biochemistry chemistry Zonula Occludens-1 Protein LLC-PK1 Cells Flux (metabolism) Tyrosine kinase Fluorescein-5-isothiocyanate Heat-Shock Response |
| Zdroj: | Journal of Cellular Physiology. 203:471-478 |
| ISSN: | 1097-4652 0021-9541 |
| DOI: | 10.1002/jcp.20234 |
| Popis: | Heat stress (HS) induces activation of high-affinity sodium-dependent glucose transporter (SGLT1) in porcine renal LLC-PK(1) cells. In this study, we investigated the roles of SGLT1 activation in reorganization of zonula occludens-1 (ZO-1), a cytosolic tight junction (TJ) protein, after HS. HS (42 degrees C, 3 h) caused decrease in transepithelial electrical resistance (TER). Subsequent incubation at 37 degrees C for 12 h increased TER above pre-HS level. The treatment of phloridzin, a potent SGLT1 inhibitor, or the replacement of glucose with a nonmetabolizable glucose analog blocked the recovery of TER and increased the transepithelial flux of FITC-dextran (4,000 Da). Immunofluorescent staining of ZO-1 showed that HS diffused ZO-1 from cell contact to cytosolic sites. Furthermore, the fraction of ZO-1 was distributed from the Triton X-100 insoluble to the Triton X-100 soluble pool. After incubation at 37 degrees C for 12 h, cell contact and ZO-1 extractability with Triton X-100 returned to pre-HS conditions, but the recovery was completely prevented by phloridzin. Tyrosine kinases activity was increased by HS that was inhibited by phloridzin. Genistein and CGP77675, tyrosine kinases inhibitors, blocked the recovery of TER and increased the transepithelial flux of FITC-dextran. Furthermore, these inhibitors prevented the recovery of cell contact and ZO-1 extractability with Triton X-100 as same as phloridzin. These findings suggested that the activation of SGLT1 reorganized ZO-1 mediated by elevation of tyrosine kinases activity after heat injury. |
| Databáze: | OpenAIRE |
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