Na+ and Ca2+ channel expression in cultured newt retinal pigment epithelial cells: comparison with neuronal types of ion channels.
We cultured retinal pigment epithelial (RPE) cells dissociated from adult newt eye and analyzed their voltage-gated ion channels during culture using whole-cell patch-clamp techniques. The results were compared with those of retinal neurons under identical experimental conditions. After 6-9 days in culture (early stage), > 60% of RPE cells developed voltage-gated Na+ and Ca2+ channels that were not observed in freshly dissociated RPE cells. The number of cells expressing Na+ channels and Na+ current density were high after 12-15 days in culture (intermediate stage), while the number of Ca2+ channel-expressing cells and Ca2+ current density were high after 20-30 days in culture (late stage). The activation voltage of the Na+ current in the RPE cells was similar to that in neurons. The voltage dependence of Na+ current inactivation was somewhat different between two cell types. The steepness of the inactivation curve tended to be less in cultured RPE cells than in neurons, and the half-inactivation voltage was about-54 mV for the RPE cells and -45 mV for neurons. The Ca2+ current expressed in cultured RPE cells was too small to detect without replacement of external Ca2+ with Ba2+. The Ba2+ current, like Ca2+ current in neurons, was enhanced by Bay-K 8644 and blocked by nicardipine. These results suggest that the RPE cells, like neurons, expressed L-type Ca2+ channels in culture. The possibility that the development of both Na2+ and Ca2+ channels in cultured RPE cells is a manifestation of the transdifferentiation of RPE cells into neurons is discussed.
Sakai
H
H
Institute of Biological Sciences, University of Tsukuba, Ibaraki, Japan.
Saito
T
T
D000818:Animals; D015220:Calcium Channels / Q000502:physiology*; D002478:Cells, Cultured; D004848:Epithelium / Q000378:metabolism; D007473:Ion Channels / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D018408:Patch-Clamp Techniques; D012160:Retina / Q000502:physiology*; D012455:Salamandridae; D015222:Sodium Channels / Q000502:physiology*
D015220:Calcium Channels; D007473:Ion Channels; D015222:Sodium Channels