CHARACTERIZATION OF ION CHANNELS SEEN IN SUBCONFLUENT HUMAN DERMAL FIBROBLASTS

1. Ion channels expressed in human dermal fibroblasts are characterized using the patch-clamp technique. 2. A number of different ion channels were found but their expression occurred at various frequencies. The most commonly found phenotype was the expression of voltage-gated K+ current. This 'typical' K+ current was seen in about 60% of the cells recorded. 3. Subtypes of voltage-gated K+ channels could be discerned by differences in gating kinetics. One has fast inactivation and resembles the 'A' K+ current. Additional subtypes were sometimes discerned based on activation kinetics. 4. The large-conductance Ca2+-activated K+ channel (maxi-K+) could be found in nearly every cell but required large depolarizations to activate using the standard Ca2+-buffered pipette solution (10(-8) M [Ca2+]i). 5. Inward rectifier K+ channels were seen in a low percentage of cells. The inward rectifier K+ current was sensitive to 'wash-out' if guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S) was included in the pipette solution dialysing the cell. 6. Tetrodotoxin (TTX)-sensitive voltage-gated Na+ channels were seen but in a lower number of cells recorded, about 20%. Evidence for subtypes of Na+ channels were sometimes seen based on differences in gating kinetics. 7. An ATP-dependent osmotically activated Cl- current was also found. This current showed some outward rectification but was otherwise voltage independent. 8. In addition, a cell-to-cell contact-associated K+ current was described. This current was linear over the voltage ranges used and whose gating correlated with the existence of gap junctions. 9. These currents were characterized to determine the baseline behaviour of unstimulated cells and to compare to bradykinin-stimulated cells described in the following paper. As unexcitable cells, human dermal fibroblasts are capable of expressing a surprising diversity of ion channel phenotypes and of ion channel modulations.