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Am J Physiol Cell Physiol 283: C223-C234, 2002. First published April 18, 2002; doi:10.1152/ajpcell.00486.2001
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Vol. 283, Issue 1, C223-C234, July 2002

Modulation of cardiac PIP2 by cardioactive hormones and other physiologically relevant interventions

Cem Nasuhoglu1, Siyi Feng1, Yanping Mao1, Imman Shammat1, Masaya Yamamato1, Svetlana Earnest2, Mark Lemmon3, and Donald W. Hilgemann1

Departments of 1 Physiology and 2 Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9040; and 3 Department of Biochemistry & Biophysics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6059

Phosphatidylinositol 4,5-bisphosphate (PIP2) affects profoundly several cardiac ion channels and transporters, and studies of PIP2-sensitive currents in excised patches suggest that PIP2 can be synthesized and broken down within 30 s. To test when, and if, total phosphatidylinositol 4-phosphate (PIP) and PIP2 levels actually change in intact heart, we used a new, nonradioactive HPLC method to quantify anionic phospholipids. Total PIP and PIP2 levels (10-30 µmol/kg wet weight) do not change, or even increase, with activation of Galpha q/phospholipase C (PLC)-dependent pathways by carbachol (50 µM), phenylephrine (50 µM), and endothelin-1 (0.3 µM). Adenosine (0.2 mM) and phorbol 12-myristate 13-acetate (1µM) both cause 30% reduction of PIP2 in ventricles, suggesting that diacylglycerol (DAG)-dependent mechanisms negatively regulate cardiac PIP2. PIP2, but not PIP, increases reversibly by 30% during electrical stimulation (2 Hz for 5 min) in guinea pig left atria; the increase is blocked by nickel (2 mM). Both PIP and PIP2 increase within 3 min in hypertonic solutions, roughly in proportion to osmolarity, and similar effects occur in multiple cell lines. Inhibitors of several volume-sensitive signaling mechanisms do not affect these responses, suggesting that PIP2 metabolism might be sensitive to membrane tension, per se.

phosphatidylinositol 4,5-bisphosphate; phosphatidylinositol; diacylglycerol; phorbol ester; cardiac muscle; G protein-coupled receptors; phospholipase C; cell volume


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