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Am J Physiol Endocrinol Metab 267: E323-E330, 1994;
0193-1849/94 $5.00
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AJP - Endocrinology and Metabolism, Vol 267, Issue 2 E323-E330, Copyright © 1994 by American Physiological Society

ARTICLES

P2-purinergic stimulation of iodide efflux in FRTL-5 rat thyroid cells involves parallel activation of PLC and PLA2

R. C. Smallridge and I. D. Gist
Department of Clinical Physiology, Walter Reed Army Institute of Research, Washington, District of Columbia 20307-5100.

Extracellular ATP increases inositol phosphates, cytosolic Ca2+ concentration ([Ca2+]i), arachidonic acid (AA) release, and iodide efflux in FRTL-5 cells. To examine the sequence of events in P2-purinergic receptor activation by ATP, a phospholipase C (PLC) inhibitor (U-73122) and a phospholipase A2 (PLA2) inhibitor (U-26384), as well as 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'- tetraacetic acid (BAPTA) and downregulation of protein kinase C (PKC) were used. ATP increased inositol trisphosphate (IP3), [Ca2+]i, AA release, and 125I efflux dose dependently. U-73122 inhibited the IP3 and calcium increase but not AA; U-26384 prevented AA release but not the increase in calcium. Both agents inhibited iodide efflux. BAPTA prevented any ATP-induced increase in [Ca2+]i without affecting AA release or 125I efflux. PKC downregulation had no effect on ATP-stimulated AA release, but reduced 125I efflux. We conclude that ATP-induced iodide efflux involves parallel, not sequential, activation of PLC and PLA2. No increase in [Ca2+]i or PKC activity is required for PLA2 activation. In contrast, an increase in 125I efflux depends on PKC and PLA2 activities, but not an increase in [Ca2+]i.


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