Caveolin-1 regulates corneal wound healing by modulating Kir4.1 activity
| Autor: | Xiao-Tong Su, Chengbiao Zhang, Lars Bellner, Dao-Hong Lin |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Pathology medicine.medical_specialty Genotype Physiology Caveolin 1 Primary Cell Culture Biology Transfection Cell Line Membrane Potentials 03 medical and health sciences 0302 clinical medicine Western blot Cell Movement medicine Animals Humans Patch clamp Phosphorylation Potassium Channels Inwardly Rectifying Corneal epithelium Mice Knockout Wound Healing medicine.diagnostic_test Cell growth Epithelium Corneal Cell migration Cell Biology Molecular biology ErbB Receptors Mice Inbred C57BL Disease Models Animal Phenotype 030104 developmental biology medicine.anatomical_structure Cell culture Call for Papers Potassium RNA Interference Wound healing 030217 neurology & neurosurgery Corneal Injuries Signal Transduction |
| Zdroj: | American Journal of Physiology-Cell Physiology. 310:C993-C1000 |
| ISSN: | 1522-1563 0363-6143 |
| DOI: | 10.1152/ajpcell.00023.2016 |
| Popis: | The expression of caveolin-1 (Cav1) in corneal epithelium is associated with regeneration potency. We used Cav1−/−mice to study the role of Cav1 in modulating corneal wound healing. Western blot and whole cell patch clamp were employed to study the effect of Cav1 deletion on Kir4.1 current density in corneas. We found that Ba2+-sensitive K+currents in primary cultured murine corneal epithelial cells (pMCE) from Cav1−/−were dramatically reduced (602 pA) compared with those from wild type (WT; 1,300 pA). As a consequence, membrane potential was elevated in pMCE from Cav1−/−compared with that from WT (−43 ± 7.5 vs. −58 ± 4.0 mV, respectively). Western blot showed that either inhibition of Cav1 expression or Ba2+incubation stimulated phosphorylation of the EGFR. The transwell migration assay showed that Cav1 genetic inactivation accelerated cell migration. The regrowth efficiency of human corneal epithelial cells (HCE) transfected with siRNA-Cav1 or negative control was evaluated by scrape injury assay. With the presence of mitomycin C (10 μg/ml) to avoid the influence of cell proliferation, Cav1 inhibition with siRNA significantly increased migration compared with control siRNA in HCE. This promoting effect by siRNA-Cav1 could not be further enhanced by cotransfection with siRNA-Kcnj10. By using corneal debridement, we found that wound healing was significantly accelerated in Cav1−/−compared with WT mice (70 ± 10 vs. 36 ± 3%, P < 0.01). Our findings imply that the mechanism by which Cav-1 knockout promotes corneal regrowth is, at least partially, due to the inhibition of Kir4.1 which stimulates EGFR signaling. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|