Purinergically induced membrane fluidization in ciliary cells: characterization and control by calcium and membrane potential.
To examine the role of membrane dynamics in transmembrane signal transduction, we studied changes in membrane fluidity in mucociliary tissues from frog palate and esophagus epithelia stimulated by extracellular ATP. Micromolar concentrations of ATP induced strong changes in fluorescence polarization, possibly indicating membrane fluidization. This effect was dosage dependent, reaching a maximum at 10-microM ATP. It was dependent on the presence of extracellular Ca2+ (or Mg2+), though it was insensitive to inhibitors of voltage-gated calcium channels. It was inhibited by thapsigargin and by ionomycin (at low extracellular Ca2+ concentration), both of which deplete Ca2+ stores. It was inhibited by the calcium-activated potassium channel inhibitors quinidine, charybdotoxin, and apamine and was reduced considerably by replacement of extracellular Na+ with K+. Hyperpolarization, or depolarization, of the mucociliary membrane induced membrane fluidization. The degree of membrane fluidization depended on the degree of hyperpolarization or depolarization of the ciliary membrane potential and was considerably lower than the effect induced by extracellular ATP. These results indicate that appreciable membrane fluidization induced by extracellular ATP depends both on an increase in intracellular Ca2+, mainly from its internal stores, and on hyperpolarization of the membrane. Calcium-dependent potassium channels couple the two effects. In light of recent results on the enhancement of ciliary beat frequency, it would appear that extracellular ATP-induced changes both in ciliary beat frequency and in membrane fluidity are triggered by similar signal transduction pathways.
Alfahel
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E
Department of Chemistry, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
Korngreen
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A
Parola
A H
AH
Priel
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Z
D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D002118:Calcium / Q000378:metabolism*; D002923:Cilia / Q000187:drug effects* / Q000378:metabolism*; D005454:Fluorescence Polarization; D066298:In Vitro Techniques; D007424:Intracellular Fluid / Q000378:metabolism; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D008560:Membrane Fluidity / Q000187:drug effects* / Q000502:physiology*; D008564:Membrane Potentials; D009079:Mucociliary Clearance / Q000502:physiology; D010159:Palate / Q000187:drug effects / Q000378:metabolism; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism; D011895:Rana ridibunda; D015398:Signal Transduction; D013552:Swine; D014132:Trachea / Q000187:drug effects / Q000378:metabolism
D007476:Ionophores; D015221:Potassium Channels; D015759:Ionomycin; D000255:Adenosine Triphosphate; D002118:Calcium