This Article Full Text Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bird, I. M. Articles by Magness, R. R. Search for Related Content PubMed PubMed Citation Articles by Bird, I. M. Articles by Magness, R. R.

Pregnancy-Dependent Changes in Cell Signaling Underlie Changes in Differential Control of Vasodilator Production in Uterine Artery Endothelial Cells¹

Perinatal Research Laboratories, Departments of Obstetrics/Gynecology (I.M.B., J.A.S., T.D., J.M.C., J.Z., R.R.M.) and Meat/Animal Science (R.R.M.), University of Wisconsin, Madison, Wisconsin 53715; and the Center for Perinatal Biology, Loma Linda University School of Medicine (L.Z.), Loma Linda, California 92350

Address all correspondence and requests for reprints to: Ian M Bird, Ph.D., Department Obstetrics and Gynecology, Perinatal Research Laboratories, University of Wisconsin, 7E Meriter Hospital/Park, 202 South Park Street, Madison, Wisconsin 53715. E-mail: imbird{at}facstaff.wisc.edu

During pregnancy, the uterine vasculature shows a marked increase in vasodilator production [prostacyclin (PGI₂) and nitric oxide (NO)] in response to a number of agonists including angiotensin II (AII) and ATP. As a consequence vascular resistance is kept low, and uterine blood flow is maximized to meet the needs of the growing fetus. Studies of the molecular basis underlying this change in control of endothelial NO and PGI₂ production have been hampered by the lack of availability of a suitable cell model. To that end we have developed and characterized a new ovine uterine artery endothelial cell (UAEC) culture model derived from nonpregnant (NP) or pregnant (P) ewes. Endothelial cells were isolated from pregnant (120–130 days; n = 6) and nonpregnant (n = 4) ewes and maintained in primary culture. Endothelial cells at passage 4 showed uniform expression of endothelial nitric oxide synthase (eNOS; an endothelial marker) as well as AII type 1 receptor and growth factor receptors and uniform uptake of acetylated low density lipoprotein (a property of endothelial cells not shared by fibroblasts or vascular smooth muscle cells), thus demonstrating cell purity. Expressions of eNOS, cyclooxygenase-1, PGI₂ synthase, cytosolic phospholipase A₂, AII type 1 receptor, and growth factor receptors are also maintained at passage 4. Mitogenesis is maintained in response to basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and vascular endothelial growth factor (VEGF) in both NP-UAEC and P-UAEC. The differential production of vasodilators by NP-UAEC and P-UAEC is maintained in a manner similar to that previously reported in vivo. Thus, P-UAEC make NO in response to AII, ATP, bFGF, EGF, and VEGF, whereas NP-UAEC make NO in response to bFGF, EGF, and VEGF only. Similarly, P-UAEC make PGI₂ in response to AII, ATP, bFGF, and VEGF, whereas NP-UAEC make PGI₂ only in response to ATP and VEGF. As both cytosolic phospholipase A₂ and eNOS may be regulated by both Ca²⁺ and protein kinases, we investigated the effects of these agonists on Ca²⁺ mobilization and ERK-1/2 phosphorylation. ATP consistently elevates Ca²⁺ levels in both P-UAEC and NP-UAEC. All other agonists were without acute (0–4 min) effect on Ca²⁺ in P-UAEC or NP-UAEC. In contrast, all agonists stimulated an acute (10 min) phosphorylation of ERK-1/2 in P-UAEC, whereas only EGF stimulated activation in NP-UAEC. P-UAEC production of PGI₂ by agonists of both heptahelical receptors and growth factor receptors correlates closely with ERK-2 phosphorylation alone. For NO, this correlation holds for heptahelical receptor agonists, but additional signaling pathways are also implicated for bFGF and VEGF. In contrast, in NP-UAEC the lack of ERK-2 phosphorylation in response to all agonists other than EGF, and the dissociation between NO or PGI₂ production and ERK-2 phosphorylation suggest that alternate pathways play a predominant role.

This article has been cited by other articles:

				K. Chang and Lubo Zhang Review Article: Steroid Hormones and Uterine Vascular Adaptation to Pregnancy Reproductive Sciences, April 1, 2008; 15(4): 336 - 348. [Abstract] [PDF]

				F. Meziani, A. Tesse, S. Welsch, H. Kremer, M. Barthelmebs, R. Andriantsitohaina, F. Schneider, and A. Gairard Expression and Biological Activity of Parathyroid Hormone-Related Peptide in Pregnant Rat Uterine Artery: Any Role for 8-Iso-Prostaglandin F2{alpha}? Endocrinology, February 1, 2008; 149(2): 626 - 633. [Abstract] [Full Text] [PDF]

				P. Brownbill, G. C. McKeeman, J. C. Brockelsby, I. P. Crocker, and C. P. Sibley Vasoactive and Permeability Effects of Vascular Endothelial Growth Factor-165 in the Term in Vitro Dually Perfused Human Placental Lobule Endocrinology, October 1, 2007; 148(10): 4734 - 4744. [Abstract] [Full Text] [PDF]

				S. M Gifford, F.-X. Yi, and I. M Bird Pregnancy-enhanced Ca2+ responses to ATP in uterine artery endothelial cells is due to greater capacitative Ca2+ entry rather than altered receptor coupling. J. Endocrinol., August 1, 2006; 190(2): 373 - 384. [Abstract] [Full Text] [PDF]

				S. M Gifford, F.-X. Yi, and I. M Bird Pregnancy-enhanced store-operated Ca2+ channel function in uterine artery endothelial cells is associated with enhanced agonist-specific transient receptor potential channel 3-inositol 1,4,5-trisphosphate receptor 2 interaction. J. Endocrinol., August 1, 2006; 190(2): 385 - 395. [Abstract] [Full Text] [PDF]

				J. A. Sullivan, M. A. Grummer, F.-X. Yi, and I. M. Bird Pregnancy-Enhanced Endothelial Nitric Oxide Synthase (eNOS) Activation in Uterine Artery Endothelial Cells Shows Altered Sensitivity to Ca2+, U0126, and Wortmannin But Not LY294002--Evidence that Pregnancy Adaptation of eNOS Activation Occurs at Multiple Levels of Cell Signaling Endocrinology, May 1, 2006; 147(5): 2442 - 2457. [Abstract] [Full Text] [PDF]

				N. I. Gokina and T. Goecks Upregulation of endothelial cell Ca2+ signaling contributes to pregnancy-enhanced vasodilation of rat uteroplacental arteries Am J Physiol Heart Circ Physiol, May 1, 2006; 290(5): H2124 - H2135. [Abstract] [Full Text] [PDF]

				J. M. Cale and I. M. Bird Dissociation of endothelial nitric oxide synthase phosphorylation and activity in uterine artery endothelial cells Am J Physiol Heart Circ Physiol, April 1, 2006; 290(4): H1433 - H1445. [Abstract] [Full Text] [PDF]

				F.-X. Yi and I. M. Bird Pregnancy-Specific Modulatory Role of Mitochondria on Adenosine 5'-Triphosphate-Induced Cytosolic [Ca2+] Signaling in Uterine Artery Endothelial Cells Endocrinology, November 1, 2005; 146(11): 4844 - 4850. [Abstract] [Full Text] [PDF]

				J. Zheng, Y. Wen, D.-b. Chen, I. M. Bird, and R. R. Magness Angiotensin II Elevates Nitric Oxide Synthase 3 Expression and Nitric Oxide Production Via a Mitogen-Activated Protein Kinase Cascade in Ovine Fetoplacental Artery Endothelial Cells Biol Reprod, June 1, 2005; 72(6): 1421 - 1428. [Abstract] [Full Text] [PDF]

				K. A. Vonnahme, M. E. Wilson, Y. Li, H. L. Rupnow, T. M. Phernetton, S. P. Ford, and R. R. Magness Circulating levels of nitric oxide and vascular endothelial growth factor throughout ovine pregnancy J. Physiol., May 15, 2005; 565(1): 101 - 109. [Abstract] [Full Text] [PDF]

				J. M. Cale, S. C. Tsoi, M. Toppe, M. A. Grummer, M. Ochiai, R. R. Magness, and I. M. Bird Molecular Cloning of Ovine Endothelial Nitric Oxide Synthase and Expression in COS-7 Cells Reproductive Sciences, April 1, 2005; 12(3): 156 - 168. [Abstract] [PDF]

				C. M. Anderson, F. Lopez, H.-Y. Zhang, K. Pavlish, and J. N. Benoit Reduced Uteroplacental Perfusion Alters Uterine Arcuate Artery Function in the Pregnant Sprague-Dawley Rat Biol Reprod, March 1, 2005; 72(3): 762 - 766. [Abstract] [Full Text] [PDF]

				W. X. Liao, R. R. Magness, and D.-b. Chen Expression of Estrogen Receptors-{alpha} and -{beta} in the Pregnant Ovine Uterine Artery Endothelial Cells In Vivo and In Vitro Biol Reprod, March 1, 2005; 72(3): 530 - 537. [Abstract] [Full Text] [PDF]

				F.-X. Yi, R. R. Magness, and I. M. Bird Simultaneous imaging of [Ca2+]i and intracellular NO production in freshly isolated uterine artery endothelial cells: effects of ovarian cycle and pregnancy Am J Physiol Regulatory Integrative Comp Physiol, January 1, 2005; 288(1): R140 - R148. [Abstract] [Full Text] [PDF]

				D. Xiao and L. Zhang Adaptation of uterine artery thick- and thin-filament regulatory pathways to pregnancy Am J Physiol Heart Circ Physiol, January 1, 2005; 288(1): H142 - H148. [Abstract] [Full Text] [PDF]

				K. A. Vonnahme and S. P. Ford Differential Expression of the Vascular Endothelial Growth Factor-Receptor System in the Gravid Uterus of Yorkshire and Meishan Pigs Biol Reprod, July 1, 2004; 71(1): 163 - 169. [Abstract] [Full Text] [PDF]

				Y. Li, J. Zheng, I. M. Bird, and R. R. Magness Mechanisms of Shear Stress-Induced Endothelial Nitric-Oxide Synthase Phosphorylation and Expression in Ovine Fetoplacental Artery Endothelial Cells Biol Reprod, March 1, 2004; 70(3): 785 - 796. [Abstract] [Full Text] [PDF]

				D.-b. Chen, I. M. Bird, J. Zheng, and R. R. Magness Membrane Estrogen Receptor-Dependent Extracellular Signal-Regulated Kinase Pathway Mediates Acute Activation of Endothelial Nitric Oxide Synthase by Estrogen in Uterine Artery Endothelial Cells Endocrinology, January 1, 2004; 145(1): 113 - 125. [Abstract] [Full Text] [PDF]

				Y. Li, J. Zheng, I. M. Bird, and R. R. Magness Effects of Pulsatile Shear Stress on Nitric Oxide Production and Endothelial Cell Nitric Oxide Synthase Expression by Ovine Fetoplacental Artery Endothelial Cells Biol Reprod, September 1, 2003; 69(3): 1053 - 1059. [Abstract] [Full Text] [PDF]

				S. M. Gifford, J. M. Cale, S. Tsoi, R. R. Magness, and I. M. Bird Pregnancy-Specific Changes in Uterine Artery Endothelial Cell Signaling in Vivo Are Both Programmed and Retained in Primary Culture Endocrinology, August 1, 2003; 144(8): 3639 - 3650. [Abstract] [Full Text] [PDF]

				I. M. Bird, L. Zhang, and R. R. Magness Possible mechanisms underlying pregnancy-induced changes in uterine artery endothelial function Am J Physiol Regulatory Integrative Comp Physiol, February 1, 2003; 284(2): R245 - R258. [Abstract] [Full Text] [PDF]

				J. M. Joyce, T. M. Phernetton, and R. R. Magness Effect of Uterine Blood Flow Occlusion on Shear Stress-Mediated Nitric Oxide Production and Endothelial Nitric Oxide Synthase Expression During Ovine Pregnancy Biol Reprod, July 1, 2002; 67(1): 320 - 326. [Abstract] [Full Text] [PDF]

				D. Xiao, X. Huang, S. Bae, C. A. Ducsay, and L. Zhang Cortisol-mediated potentiation of uterine artery contractility: effect of pregnancy Am J Physiol Heart Circ Physiol, July 1, 2002; 283(1): H238 - H246. [Abstract] [Full Text] [PDF]

				T. Kimura, K. Ayajiki, Y. Noda, and T. Okamura Comparison of the Responses to Thrombin in Monkey Renal and Uterine Arteries Reproductive Sciences, May 1, 2002; 9(3): 146 - 151. [Abstract] [PDF]

				H. L. Rupnow, T. M. Phernetton, M. L. Modrick, M. C. Wiltbank, I. M. Bird, and R. R. Magness Endothelial Vasodilator Production by Uterine and Systemic Arteries. VIII. Estrogen and Progesterone Effects on cPLA2, COX-1, and PGIS Protein Expression Biol Reprod, February 1, 2002; 66(2): 468 - 474. [Abstract] [Full Text] [PDF]

				D. Xiao and L. Zhang ERK MAP kinases regulate smooth muscle contraction in ovine uterine artery: effect of pregnancy Am J Physiol Heart Circ Physiol, January 1, 2002; 282(1): H292 - H300. [Abstract] [Full Text] [PDF]

				J. M. Joyce, T. M. Phernetton, C. E. Shaw, M. L. Modrick, and R. R. Magness Endothelial vasodilator production by uterine and systemic arteries. IX. eNOS gradients in cycling and pregnant ewes Am J Physiol Heart Circ Physiol, January 1, 2002; 282(1): H342 - H348. [Abstract] [Full Text] [PDF]

				J. K. Peterson, F. Moran, A. J. Conley, and I. M. Bird Zonal Expression of Endothelial Nitric Oxide Synthase in Sheep and Rhesus Adrenal Cortex Endocrinology, December 1, 2001; 142(12): 5351 - 5363. [Abstract] [Full Text] [PDF]

				T. Di, J. A. Sullivan, R. R. Magness, L. Zhang, and I. M. Bird Pregnancy-Specific Enhancement of Agonist-Stimulated ERK-1/2 Signaling in Uterine Artery Endothelial Cells Increases Ca2+ Sensitivity of Endothelial Nitric Oxide Synthase as well as Cytosolic Phospholipase A2 Endocrinology, July 1, 2001; 142(7): 3014 - 3026. [Abstract] [Full Text] [PDF]

				D. Xiao, W. J. Pearce, and L. Zhang Pregnancy enhances endothelium-dependent relaxation of ovine uterine artery: role of NO and intracellular Ca2+ Am J Physiol Heart Circ Physiol, July 1, 2001; 281(1): H183 - H190. [Abstract] [Full Text] [PDF]

				R. R. Magness, J. A. Sullivan, Y. Li, T. M. Phernetton, and I. M. Bird Endothelial vasodilator production by uterine and systemic arteries. VI. Ovarian and pregnancy effects on eNOS and NOx Am J Physiol Heart Circ Physiol, April 1, 2001; 280(4): H1692 - H1698. [Abstract] [Full Text] [PDF]

				H. L. Rupnow, T. M. Phernetton, C. E. Shaw, M. L. Modrick, I. M. Bird, and R. R. Magness Endothelial vasodilator production by uterine and systemic arteries. VII. Estrogen and progesterone effects on eNOS Am J Physiol Heart Circ Physiol, April 1, 2001; 280(4): H1699 - H1705. [Abstract] [Full Text] [PDF]

				L. P. Reynolds and D. A. Redmer Angiogenesis in the Placenta Biol Reprod, April 1, 2001; 64(4): 1033 - 1040. [Abstract] [Full Text]

				D. Xiao, I. M. Bird, R. R. Magness, L. D. Longo, and L. Zhang Upregulation of eNOS in pregnant ovine uterine arteries by chronic hypoxia Am J Physiol Heart Circ Physiol, February 1, 2001; 280(2): H812 - H820. [Abstract] [Full Text] [PDF]

				C. Kurz, L. Hefler, H. Zeisler, C. Schatten, P. Husslein, and C. Tempfer Maternal Basic Fibroblast Growth Factor Serum Levels Are Associated With Pregnancy-Induced Hypertension Reproductive Sciences, January 1, 2001; 8(1): 24 - 26. [Abstract] [PDF]