Sequential activation of store-operated currents in human gingival keratinocytes.
Calcium ion store-activated currents in undifferentiated human gingival keratinocytes were measured with the whole cell patch clamp and fura techniques. Thapsigargin or intracellular inositol 1,4,5-trisphosphate and BAPTA rapidly induced an early transient current with I(CRAC) (calcium release activated calcium ion current) characteristics, and several later, larger sustained currents that depended on the mode of store depletion. Thapsigargin activated two currents within minutes of I(CRAC) activation. The first was a nonspecific cation current, I(NSC). A second conducted Na+ and Cs+, and was partially inhibited by thapsigargin (INa1). Dialysis with inositol 1,4,5-trisphosphate and BAPTA induced a later current that also conducted Na+ and Cs+, but was inhibited by extracellular calcium ion (INa2), with properties consistent with an epithelial Na+ channel current in some cells, and a calcium ion-insensitive Na+ current (INa3). Comparison of thapsigargin-evoked current changes with fura-2/AM results from separate cells indicated that both the I(CRAC) and the later, larger calcium ion conducting currents contributed to changes in intracellular calcium ion concentration, and likely play important parts in calcium ion signaling in undifferentiated keratinocytes.
Fatherazi
Sahba
S
Department of Oral Biology, School of Dentistry, University of Washington, Seattle 98195, USA.
Belton
Carol M
CM
Izutsu
Kenneth T
KT
D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D004533:Egtazic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005881:Gingiva / Q000166:cytology*; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000494:pharmacology; D015603:Keratinocytes / Q000166:cytology / Q000502:physiology*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D018408:Patch-Clamp Techniques; D012964:Sodium / Q000378:metabolism; D019284:Thapsigargin / Q000494:pharmacology
D015220:Calcium Channels; D002614:Chelating Agents; D004791:Enzyme Inhibitors; D004533:Egtazic Acid; D019284:Thapsigargin; D015544:Inositol 1,4,5-Trisphosphate; D012964:Sodium; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium