Effect of purinergic stimulation on intracellular calcium concentration and transepithelial potential difference in cultured bovine oviduct cells.
Epithelial cells were removed from bovine oviducts by enzyme digestion and either cultured on laminin-coated coverslips (for determination of [Ca2+]i) or on collagen filters (for determination of transepithelial potential difference [pd]). Cells on coverslips were loaded with Fura-2 to monitor [Ca2+]i. Application of extracellular ATP induced a transient increase in [Ca2+]i in a dose-dependent manner. This response was abolished by thapsigargin, indicating that the rise in [Ca2+]i was derived from intracellular stores. The order of potency of the nucleotide-induced rise in [Ca2+]i was uridine triphosphate (UTP)>ATP>ADP. Epithelial cells were grown on collagen filters, and when mounted in a modified Ussing chamber exhibited an electrical pd of 1.00 +/- 0.36 mV with the apical side negative with respect to the basal. Application of UTP, ATP, and ADP to the basal side induced transient increases in pd of 1.15 +/- 0.21, 0.77 +/- 0.16, and 0.26 +/- 0.06 mV, respectively. The order of potency of the nucleotides in eliciting transient increases in [Ca2+]i and pd suggests the presence of a P2u purinergic receptor in the bovine oviduct epithelium that could play a role in transepithelial ion movements and hence the control of oviductal fluid formation.
Cox
C I
CI
Department of Biology, University of York, Heslington, United Kingdom.
Leese
H J
HJ
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D004848:Epithelium / Q000187:drug effects / Q000502:physiology; D005187:Fallopian Tubes / Q000187:drug effects / Q000502:physiology*; D005260:Female; D005455:Fluorescent Antibody Technique; D008564:Membrane Potentials; D011687:Purines / Q000494:pharmacology*; D013729:Terpenes / Q000494:pharmacology; D019284:Thapsigargin; D014544:Uridine Triphosphate / Q000494:pharmacology
D011687:Purines; D013729:Terpenes; D000244:Adenosine Diphosphate; D019284:Thapsigargin; D000255:Adenosine Triphosphate; D002118:Calcium; D014544:Uridine Triphosphate