title: The loss of sustained Ca(2+) signaling underlies suppressed endothelial nitric oxide production in preeclamptic pregnancies: implications for new therapy.
abstract: Approximately 8% of pregnancies are complicated by preeclampsia (PE), a hypertensive condition characterized by widespread endothelial dysfunction. Reduced nitric oxide (NO) output in PE subjects has been inferred but not directly measured, and there is little understanding of why this occurs. To address this we have used direct imaging of changes in intracellular Ca(2+) concentration ([Ca(2+)]i) and NO in umbilical vein endothelium of normal and PE subjects that is still intact and on the vessel luminal surface. This was achieved by dissection and preloading with fura 2 and DAF-2 imaging dyes, respectively, before subsequent challenge with ATP (100 μM, 30 min). As a control to reveal the content of active endothelial nitric oxide synthase (eNOS) per vessel segment, results were compared with a maximal stimulus with ionomycin (5 μM, 30 min). We show for the first time that normal umbilical vein endothelial cells respond to ATP with sustained bursting that parallels sustained NO output. Furthermore, in subjects with PE, a failure of sustained [Ca(2+)]i bursting occurs in response to ATP and is associated with blunted NO output. In contrast, NO responses to maximal [Ca(2+)]i elevation using ionomycin and the levels of eNOS protein are more similar between groups than the responses to ATP. When the endothelial cells from PE subjects are isolated and allowed to recover in culture, they regain the ability under fura 2 imaging to show multiple [Ca(2+)]i bursts otherwise seen in the cells from normal subjects. Thus novel clinical therapy aimed at restoring function in vivo may be possible.
authors.affiliation: Perinatal Research Laboratories, Department of Obstetrics and Gynecology, School of Medicine and Public Health, University of Wisconsin Madison, Madison, Wisconsin; and.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Krupp
authors.forename: Derek S
authors.initials: DS
authors.lastname: Boeldt
authors.forename: Fu-Xian
authors.initials: FX
authors.lastname: Yi
authors.forename: Mary A
authors.initials: MA
authors.lastname: Grummer
authors.forename: Heather A
authors.initials: HA
authors.lastname: Bankowski Anaya
authors.forename: Dinesh M
authors.initials: DM
authors.lastname: Shah
authors.forename: Ian M
authors.initials: IM
authors.lastname: Bird
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000293:Adolescent; D000328:Adult; D061207:Calcium Ionophores / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects; D016022:Case-Control Studies; D002478:Cells, Cultured; D015536:Down-Regulation; D005260:Female; D005456:Fluorescent Dyes; D016257:Fura-2 / Q000031:analogs & derivatives; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000379:methods; D009569:Nitric Oxide / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D011225:Pre-Eclampsia / Q000378:metabolism* / Q000503:physiopathology / Q000628:therapy; D011247:Pregnancy; D013997:Time Factors; D055815:Young Adult
chemical_list: D061207:Calcium Ionophores; D005456:Fluorescent Dyes; C049925:fura-2-am; D009569:Nitric Oxide; D015759:Ionomycin; D000255:Adenosine Triphosphate; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D016257:Fura-2
keywords: calcium ion; endothelium; hypertension
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