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Am J Physiol Cell Physiol 257: C1020-C1029, 1989;
0363-6143/89 $5.00
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AJP - Cell Physiology, Vol 257, Issue 5 C1020-C1029, Copyright © 1989 by American Physiological Society

ARTICLES

Intracellular [Ca2+] and inositol phosphates in avian nasal gland cells

T. J. Shuttleworth and J. L. Thompson
Department of Physiology, University of Rochester School of Medicine and Dentistry, New York 14642.

Isolated cells from the nasal salt gland of ducklings (Anas platyrhynchos) were evaluated as a model system for the study of the muscarinic activation of exocrine ion secretion. Cells loaded with the fluorescent probe indo-1 were used to study changes in intracellular Ca2+ concentration [( Ca2+]i) after stimulation. Changes in inositol phosphate generation and oxygen consumption were also determined. Loading with the acetomethoxy ester form of indo-1 (indo-1/AM) was rapid, and intracellular cleavage of the ester was essentially complete. Leakage of the dye was negligible over the time course of measurements (up to 20 min). Resting [Ca2+]i was approximately 100 nM. Stimulation with carbachol resulted in progressive increases in the generation of inositol phosphates and rapid four- to fivefold increases in [Ca2+]i. At normal extracellular Ca2+ concentrations, [Ca2+]i remained elevated (approximately 3 times resting levels) for as long as stimulation continued. Experiments showed that the increases in [Ca2+]i were comprised of a combination of release of Ca2+ from intracellular stores and an enhanced entry of Ca2+ from the extracellular medium. It is specifically this latter process that produces the sustained elevations in [Ca2+]i that are the essential signal for secretory activity.


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