We investigated the mechanism of phospholipase A 2 (PLA 2 ) activation in response to the P2 receptor agonist ATP in rat thyroid FRTL-5 cells. The PLA 2 activity was determined by measuring the release of [ 3 H]-arachidonic acid (AA) from prelabeled cells. ATP evoked a dose-and time-dependent AA rel
ATP activates a Ca2+-dependent Cl−current in the rat thyroid cell line, FRTL-5
✍ Scribed by S. C. Martin
- Publisher
- Springer
- Year
- 1992
- Tongue
- English
- Weight
- 922 KB
- Volume
- 125
- Category
- Article
- ISSN
- 0022-2631
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✦ Synopsis
The effect of external ATP on both the membrane potential and the transmembrane current of the thyroid cell line FRTL-5 has been investigated in the patch-clamp whole-cell recording configuration. In the resting situation the membrane potential is around -70 mV and the membrane acts like a K(+)-sensitive electrode. Application of ATP at concentrations higher than 1 microM elicited an increase in Cl- conductance, responsible for a membrane depolarization which could be blocked by preincubation with the P2-antagonist quinidine. Chelation of intracellular Ca2+ also blocked the ATP induced changes in membrane potential and Cl- current. Intracellular perfusion with inositol trisphosphate (IP3) (50 microM) also stimulated a Cl- current which mimicked the response induced by ATP. ATP is able to initiate a response in the absence of extracellular Ca2+, but also opens a Ca(2+)-influx pathway, as demonstrated by a secondary response upon Ca2+ readmission in the external medium, in the continued presence of ATP. ADP and ATP gamma S were able to mimic the ATP response, whereas AMP and adenosine were unable to elicit a Cl- current. The P2X receptor agonist alpha,beta-methyleneATP was without effect as was the P2Y receptor agonist 2-methylthio ATP. We conclude that ATP is able to elicit a large IP3-mediated Ca(2+)-dependent Cl- current and membrane depolarization via a novel P2-type purinergic receptor.
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