Adenosine triphosphate activates the phospholipase-C cascade system in human amnion cells without increasing prostaglandin production.
Human amnion is hypothesized to be a target tissue for hormone messages from the fetus regarding labor. We have previously demonstrated prostaglandin E2 (PGE2) release in amnion after treatment with phorbol and oxytocin, but other potential agonists of the inositol phospholipid/protein kinase-C system have not been investigated. The effects of extracellular ATP on cytosolic calcium concentration [( Ca2+])i) inositol phosphate (IP) accumulation, and PGE2 production were studied in cultured human amnion cells. Intracellular free calcium [Ca2+]i was measured using the fluorescent dye fura-2. Addition of 0.01-30 microM ATP resulted in a [Ca2+]i transient which peaked within 15 sec and returned to baseline over 10 min. UTP (1 microM) was more effective than ATP (1 microM); [Ca2+]i levels rose from 233 to 2880 nM (UTP) and 2320 nM (ATP). A reduced effect was observed with other nucleotides in a rank order of agonist potency of ITP greater than CTP greater than ADP greater than GTP greater than TTP. No effect was seen with AMP, cAMP, or adenosine. This is consistent with P2 purinoceptors, as described in other tissues. ATP (100 microM) also dramatically increased IP accumulation. Inositol triphosphate, inositol bisphosphate, and inositol monophosphate were increased 7-, 9-, and 16-fold respectively. The agonist potency order of other nucleotides for IP accumulation was the same as that of [Ca2+]i. Pharmacological concentrations of ATP (1 mM) were required to increase PGE2 production. Many other nucleotides were equally effective at this concentration. ATP activates the phospholipase-C system in human amnion, as demonstrated by the increase in [Ca2+]i and inositol phosphates. The physiological significance of purinergic stimulation of this tissue remains unclear.
Vander Kooy
D
D
Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, Ohio 44109.
Dubyak
G R
GR
Moore
R M
RM
Moore
J J
JJ
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000650:Amnion / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D002118:Calcium / Q000032:analysis / Q000378:metabolism; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000187:drug effects; D004848:Epithelium / Q000032:analysis; D005260:Female; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism; D011247:Pregnancy; D011453:Prostaglandins / Q000378:metabolism*; D010738:Type C Phospholipases / Q000378:metabolism*
D007295:Inositol Phosphates; D011453:Prostaglandins; D000255:Adenosine Triphosphate; D010738:Type C Phospholipases; D002118:Calcium