期刊
JOURNAL OF GENERAL PHYSIOLOGY
卷 127, 期 3, 页码 237-251出版社
ROCKEFELLER UNIV PRESS
DOI: 10.1085/jgp.200509366
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资金
- NHLBI NIH HHS [R01 HL046764-11, HL46764, HL65435, R01 HL046764-12, R01 HL046764, R01 HL065435-04, R01 HL065435] Funding Source: Medline
Stretch of beta 1 integrins activates an outwardly rectifying, tamoxifen-sensitive Cl- current (Cl- SAC) via AT1 receptors, NADPH oxidase, and reactive oxygen species, and Cl- SAC resembles the volume-sensitive Cl- current (I-Cl,I-swell). Epidermal growth factor receptor (EGFR) kinase undergoes transactivation upon stretch, integrin engagement, and AT1 receptor activation and, in turn, stimulates NADPH oxidase. Therefore, we tested whether Cl- SAC is regulated by EGFR kinase signaling and is volume sensitive. Paramagnetic beads coated with mAb for beta 1 integrin were attached to myocytes and pulled with an electromagnet. Stretch activated a Cl- SAC that was 1.13 +/- 0.10 pA/pF at + 40 mV. AG1478 (10 mu M), an EGFR kinase blocker, inhibited 93 +/- 13% of Cl- SAC, and intracellular pretreatment with 1 mu M AG1478 markedly suppressed Cl- SAC activation. EGF (3.3 nM) directly activated an outwardly rectifying Cl- current (0.81 +/- 0.05 pA/ pF at + 40 mV) that was fully blocked by 10 mu M tamoxifen, an ICl, swell blocker. Phosphatidylinositol 3-kinase (PI-3K) is downstream of EGFR kinase. Wortmannin ( 500 nM) and LY294002 (100 mu M), blockers of PI-3K, inhibited Cl- SAC by 67 +/- 6% and 91 +/- 25% respectively, and the EGF-induced Cl- current also was fully blocked by LY294002. Furthermore, gp91ds-tat (500 nM), a cell-permeable, chimeric peptide that specifically blocks NADPH oxidase assembly, profoundly inhibited the EGF-induced Cl- current. Inactive permeant and active impermeant control peptides had no effect. Myocyte shrinkage with hyperosmotic bathing media inhibited the Cl- SAC and EGF-induced Cl- current by 88 +/- 9% and 127 +/- 11%, respectively. These results suggest that beta 1 integrin stretch activates Cl- SAC via EGFR, PI-3K, and NADPH oxidase, and that both the Cl- SAC and the EGF-induced Cl- currents are likely to be the volume-sensitive Cl- current, I-Cl,I-swell.
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