American Journal of Physiology: Cell Physiology, 2011-06, Vol.300 (6), p.C1246-C1259
2011
Details
- Autor(en) / Beteiligte
- Titel
- Interaction of ASIC1 and ENaC subunits in human glioma cells and rat astrocytes
- Ist Teil von
- American Journal of Physiology: Cell Physiology, 2011-06, Vol.300 (6), p.C1246-C1259
- Ort / Verlag
- United States: American Physiological Society
- Erscheinungsjahr
- 2011
- Link zum Volltext
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- Glioblastoma multiforme (GBM) is the most common and aggressive of the primary brain tumors. These tumors express multiple members of the epithelial sodium channel (ENaC)/degenerin (Deg) family and are associated with a basally active amiloride-sensitive cation current. We hypothesize that this glioma current is mediated by a hybrid channel composed of a mixture of ENaC and acid-sensing ion channel (ASIC) subunits. To test the hypothesis that ASIC1 interacts with αENaC and γENaC at the cellular level, we have used total internal reflection fluorescence microscopy (TIRFM) in live rat astrocytes transiently cotransfected with cDNAs for ASIC1-DsRed plus αENaC-yellow fluorescent protein (YFP) or ASIC1-DsRed plus γENaC-YFP. TIRFM images show colocalization of ASIC1 with both αENaC and γENaC. Furthermore, using TIRFM in stably transfected D54-MG cells, we also found that ASIC1 and αENaC both localize to a submembrane region following exposure to pH 6.0, similar to the acidic conditions found in the core of a glioblastoma lesion. Using high-resolution clear native gel electrophoresis, we found that ASIC1 forms a complex with ENaC subunits which migrates at ≈480 kDa in D54-MG glioma cells. These data suggest that different ENaC/Deg subunits interact and could combine to form a hybrid channel that likely underlies the amiloride-sensitive current seen in human glioma cells.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0363-6143eISSN: 1522-1563DOI: 10.1152/ajpcell.00199.2010Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3118626
- Format
- –
- Schlagworte
- Acid Sensing Ion Channels, Animals, Astrocytes - cytology, Astrocytes - metabolism, Brain, Brain Neoplasms - metabolism, Brain Neoplasms - pathology, Cell adhesion & migration, Cell Line, Tumor, Cells, Epithelial Sodium Channels - genetics, Epithelial Sodium Channels - metabolism, Glioma - metabolism, Glioma - pathology, Humans, Luminescent Proteins - genetics, Luminescent Proteins - metabolism, Membrane Transporters, Ion Channels and Pumps, Nerve Tissue Proteins - genetics, Nerve Tissue Proteins - metabolism, Patch-Clamp Techniques, Protein Isoforms - genetics, Protein Isoforms - metabolism, Proteins, Rats, Rats, Sprague-Dawley, Recombinant Fusion Proteins - genetics, Recombinant Fusion Proteins - metabolism, Rodents, Sodium, Sodium Channels - genetics, Sodium Channels - metabolism, Tumors