Circulation Research, Vol 72, 827-836, Copyright © 1993 by American Heart Association

ARTICLES

Flow-related responses of intracellular inositol phosphate levels in cultured aortic endothelial cells

AR Prasad, SA Logan, RM Nerem, CJ Schwartz and EA Sprague
Department of Pathology, University of Texas Health Science Center, San Antonio 78284-7800.

In vitro and in vivo evidence indicates that hemodynamic wall shear stress evokes a diversity of biological responses in vascular endothelial cells, ranging from cell shape changes to alterations in low density lipoprotein receptor expression. The signal transduction mechanisms by which the level of fluid mechanical shear stress is recognized by the endothelial cell and translated into these diverse biological responses remain to be elucidated. The present study focuses on the association between the onset of elevated shear stress and activation of the phosphoinositide signal transduction pathway, as measured by the intracellular release of inositol phosphates, in cultured bovine aortic endothelial cells (BAECs). BAECs were seeded, grown to confluence on large polyester sheets, and preincubated with 0.3 microCi/ml [3H]inositol for 24 hours before insertion in parallel- plate flow chambers for exposure to high shear stress (HS) at 30 dynes/cm2 or low shear stress (LS) at < 0.5 dyne/cm2 for periods ranging from 15 seconds to 24 hours. The induction of HS was associated with an early, transient but significant increase (142%, HS/LS x 100%) in inositol trisphosphate (IP3) measured at 15 seconds of shear stress exposure followed by a major peak in IP3 (189%) observed at 5 minutes after HS onset. After these initial increases, IP3 levels returned to near resting levels within 30 minutes of continued HS exposure and then continued to decline to significantly lower (75%) levels relative to LS- treated cells within 4 hours and remained lower throughout the remainder of the 24-hour HS exposure. LS-treated cells exhibited no significant changes in inositol phosphate levels throughout the 24-hour exposure periods. Exposure of BAECs to shear stress of 60 dynes/cm2 resulted in an approximately fourfold increase in IP3 levels (396%) measured at 5 minutes, almost double the levels measured in cells exposed to 30 dynes/cm2 for 5 minutes. Pretreatment of BAECs for 30 minutes with 5 mM neomycin, an inhibitor of phosphoinositide metabolism, before HS exposure inhibited both the early increases in inositol phosphates and subsequent cell elongation and alignment observed in untreated BAECs simultaneously exposed to HS without inhibiting protein synthesis. These results indicate that the exposure of cultured BAECs to elevated wall shear stress is associated with an early biphasic IP3 increase followed by a resetting of intracellular inositol phosphate concentrations to levels below that observed in static cultured BAECs. Furthermore, neomycin inhibition of this IP3 response to shear stress is associated with an inhibition of one of the major endothelial biological responses to shear stress, i.e., cell shape change and orientation.(ABSTRACT TRUNCATED AT 400 WORDS)

This article has been cited by other articles:

				A. J. Siller-Jackson, S. Burra, S. Gu, X. Xia, L. F. Bonewald, E. Sprague, and J. X. Jiang Adaptation of Connexin 43-Hemichannel Prostaglandin Release to Mechanical Loading J. Biol. Chem., September 26, 2008; 283(39): 26374 - 26382. [Abstract] [Full Text] [PDF]

				J. T. Butcher, A. M. Penrod, A. J. Garcia, and R. M. Nerem Unique Morphology and Focal Adhesion Development of Valvular Endothelial Cells in Static and Fluid Flow Environments Arterioscler Thromb Vasc Biol, August 1, 2004; 24(8): 1429 - 1434. [Abstract] [Full Text] [PDF]

				P. P. Cherian, B. Cheng, S. Gu, E. Sprague, L. F. Bonewald, and J. X. Jiang Effects of Mechanical Strain on the Function of Gap Junctions in Osteocytes Are Mediated through the Prostaglandin EP2 Receptor J. Biol. Chem., October 31, 2003; 278(44): 43146 - 43156. [Abstract] [Full Text] [PDF]

				H.-Y. Kwan, P.-C. Leung, Y. Huang, and X. Yao Depletion of Intracellular Ca2+ Stores Sensitizes the Flow-Induced Ca2+ Influx in Rat Endothelial Cells Circ. Res., February 21, 2003; 92(3): 286 - 292. [Abstract] [Full Text] [PDF]

				J. H. Hoger, V. I. Ilyin, S. Forsyth, and A. Hoger Shear stress regulates the endothelial Kir2.1 ion channel PNAS, May 15, 2002; (2002) 102184999. [Abstract] [Full Text] [PDF]

				B. Cheng, Y. Kato, S. Zhao, J. Luo, E. Sprague, L. F. Bonewald, and J. X. Jiang PGE2 Is Essential for Gap Junction-Mediated Intercellular Communication between Osteocyte-Like MLO-Y4 Cells in Response to Mechanical Strain Endocrinology, August 1, 2001; 142(8): 3464 - 3473. [Abstract] [Full Text] [PDF]

				T. Osanai, N. Akutsu, N. Fujita, T. Nakano, K. Takahashi, W. Guan, and K. Okumura Cross talk between prostacyclin and nitric oxide under shear in smooth muscle cell: role in monocyte adhesion Am J Physiol Heart Circ Physiol, July 1, 2001; 281(1): H177 - H182. [Abstract] [Full Text] [PDF]

				Q.-K. Tran, K. Ohashi, and H. Watanabe Calcium signalling in endothelial cells Cardiovasc Res, October 1, 2000; 48(1): 13 - 22. [Abstract] [Full Text] [PDF]

				M. A. Haidekker, N. L'Heureux, and J. A. Frangos Fluid shear stress increases membrane fluidity in endothelial cells: a study with DCVJ fluorescence Am J Physiol Heart Circ Physiol, April 1, 2000; 278(4): H1401 - H1406. [Abstract] [Full Text] [PDF]

				T. Osanai, N. Fujita, N. Fujiwara, T. Nakano, K. Takahashi, W. Guan, and K. Okumura Cross talk of shear-induced production of prostacyclin and nitric oxide in endothelial cells Am J Physiol Heart Circ Physiol, January 1, 2000; 278(1): H233 - H238. [Abstract] [Full Text] [PDF]

				J. J. Chiu, B. S. Wung, H. J. Hsieh, L. W. Lo, and D. L. Wang Nitric Oxide Regulates Shear Stress–Induced Early Growth Response-1 : Expression via the Extracellular Signal–Regulated Kinase Pathway in Endothelial Cells Circ. Res., August 6, 1999; 85(3): 238 - 246. [Abstract] [Full Text] [PDF]

				K.-D. Chen, Y.-S. Li, M. Kim, S. Li, S. Yuan, S. Chien, and J. Y-J. Shyy Mechanotransduction in Response to Shear Stress. ROLES OF RECEPTOR TYROSINE KINASES, INTEGRINS, AND Shc J. Biol. Chem., June 25, 1999; 274(26): 18393 - 18400. [Abstract] [Full Text] [PDF]

				S. Mohan, N. Mohan, A. J. Valente, and E. A. Sprague Regulation of low shear flow-induced HAEC VCAM-1 expression and monocyte adhesion Am J Physiol Cell Physiol, May 1, 1999; 276(5): C1100 - C1107. [Abstract] [Full Text] [PDF]

				J. M. Muller, M. J. Davis, L. Kuo, and W. M. Chilian Changes in coronary endothelial cell Ca2+ concentration during shear stress- and agonist-induced vasodilation Am J Physiol Heart Circ Physiol, May 1, 1999; 276(5): H1706 - H1714. [Abstract] [Full Text] [PDF]

				S.J. Millward-Sadler, M.O. Wright, H.-S. Lee, K. Nishida, H. Caldwell, G. Nuki, and D.M. Salter Integrin-regulated Secretion of Interleukin 4: A Novel Pathway of Mechanotransduction in Human Articular Chondrocytes J. Cell Biol., April 5, 1999; 145(1): 183 - 189. [Abstract] [Full Text] [PDF]

				C. Cardillo, C. M. Kilcoyne, R. O. Cannon III, and J. A. Panza Impairment of the nitric oxide-mediated vasodilator response to mental stress in hypertensive but not in hypercholesterolemic patients J. Am. Coll. Cardiol., November 1, 1998; 32(5): 1207 - 1213. [Abstract] [Full Text] [PDF]

				K. Matrougui, P. Schiavi, D. Guez, and D. Henrion High Sodium Intake Decreases Pressure-Induced (Myogenic) Tone and Flow-Induced Dilation in Resistance Arteries From Hypertensive Rats Hypertension, July 1, 1998; 32(1): 176 - 179. [Abstract] [Full Text] [PDF]

				H. Watanabe, R. Takahashi, X.-x. Zhang, Y. Goto, H. Hayashi, J. Ando, M. Isshiki, M. Seto, H. Hidaka, I. Niki, et al. An essential role of myosin light-chain kinase in the regulation of agonist- and fluid flow-stimulated Ca2+ influx in endothelial cells FASEB J, March 1, 1998; 12(3): 341 - 348. [Abstract] [Full Text]

				J.J. Chiu, B.S. Wung, J. Y.J. Shyy, H.J. Hsieh, and D.L. Wang Reactive Oxygen Species Are Involved in Shear Stress-Induced Intercellular Adhesion Molecule-1 Expression in Endothelial Cells Arterioscler Thromb Vasc Biol, December 1, 1997; 17(12): 3570 - 3577. [Abstract] [Full Text]

				A. M. Malek, I. W. Lee, S. L. Alper, and S. Izumo Regulation of endothelin-1 gene expression by cell shape and the microfilament network in vascular endothelium Am J Physiol Cell Physiol, November 1, 1997; 273(5): C1764 - C1774. [Abstract] [Full Text] [PDF]

				J. S. Mondy, V. Lindner, J. K. Miyashiro, B. C. Berk, R. H. Dean, and R. L. Geary Platelet-Derived Growth Factor Ligand and Receptor Expression in Response to Altered Blood Flow In Vivo Circ. Res., September 19, 1997; 81(3): 320 - 327. [Abstract] [Full Text]

				B. Hogers, M.C. DeRuiter, A.C. Gittenberger-de Groot, and R.E. Poelmann Unilateral Vitelline Vein Ligation Alters Intracardiac Blood Flow Patterns and Morphogenesis in the Chick Embryo Circ. Res., April 19, 1997; 80(4): 473 - 481. [Abstract] [Full Text]

				T. F. Wiesner, B. C. Berk, and R. M. Nerem A mathematical model of the cytosolic-free calcium response in endothelial cells to fluid shear stress PNAS, April 15, 1997; 94(8): 3726 - 3731. [Abstract] [Full Text] [PDF]

				S. R.P. Gudi, C. B. Clark, and J. A. Frangos Fluid Flow Rapidly Activates G Proteins in Human Endothelial Cells: Involvement of G Proteins in Mechanochemical Signal Transduction Circ. Res., October 1, 1996; 79(4): 834 - 839. [Abstract] [Full Text]

				K. Ayajiki, M. Kindermann, M. Hecker, I. Fleming, and R. Busse Intracellular pH and Tyrosine Phosphorylation but Not Calcium Determine Shear Stress–Induced Nitric Oxide Production in Native Endothelial Cells Circ. Res., May 1, 1996; 78(5): 750 - 758. [Abstract] [Full Text]

				A. Malek and S Izumo Mechanism of endothelial cell shape change and cytoskeletal remodeling in response to fluid shear stress J. Cell Sci., January 4, 1996; 109(4): 713 - 726. [Abstract] [PDF]

				P. L. Walpola, A. I. Gotlieb, M. I. Cybulsky, and B. L. Langille Expression of ICAM-1 and VCAM-1 and Monocyte Adherence in Arteries Exposed to Altered Shear Stress Arterioscler Thromb Vasc Biol, January 1, 1995; 15(1): 2 - 10. [Abstract] [Full Text]