R. Magness | University of Wisconsin-Madison (original) (raw)

Papers by R. Magness

Journal of Biological Chemistry, 1990

Biology of Reproduction, 2011

Biology of Reproduction, 1992

Biology of Reproduction, Jul 1, 2011

Endocrinology, 1999

Basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and vascular endothelial gr... more Basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and vascular endothelial growth factor (VEGF) may play important roles in the placental vasculature, not only by controlling cell growth and differentiation, but also by mediating production of local vasodilators such as nitric oxide. As the mitogen-activated protein kinase (MAPK) signal cascade has been widely associated with cell growth in response to growth factors, herein we investigate whether bFGF, EGF, and VEGF also stimulate expression of endothelial nitric oxide synthase (eNOS) via activation of the MAPK cascade in ovine fetoplacental artery endothelial cells. The presence of the receptors for all three growth factors was confirmed by both immunocytochemistry and a functional cell proliferation assay. All three growth factors at 10 ng/ml rapidly (<10 min) activated MAPK. This activation was inhibited by PD 98059, a specific MAPK kinase inhibitor. bFGF and EGF, but not VEGF, dose- and time-dependently i...

Reproductive Sciences, 2018

Nitric oxide (NO) production is essential to facilitate rises in uterine blood flow (UBF) during ... more Nitric oxide (NO) production is essential to facilitate rises in uterine blood flow (UBF) during pregnancy. It has been proposed that the metabolites of E 2 b, 2-hydroxyestradiol (2-OHE 2), 4-hydroxyestradiol (4-OHE 2), 2-methoxyestradiol (2-ME 2), and 4-methoxyestradiol (4-ME 2) play a role in mediating vasodilation and rises in UBF during pregnancy. We previously showed that the E 2 b metabolites stimulate prostacyclin production in pregnancy-derived ovine uterine artery endothelial cells (P-UAECs); however, it is unknown whether the E 2 b metabolites also induce NO production. Herein, UAECs derived from nonpregnant and pregnant ewes were used to test the hypothesis that E 2 b metabolites stimulate NO production in a pregnancy-specific manner. Specific estrogen receptor (ER) and adrenergic receptor (AR) antagonists were used to determine the roles of ERs or ARs in E 2 b metabolite-induced NO production. E 2 b and its metabolites increased total nitric oxide metabolites (NOx) levels (NO 2 þ NO 3) in P-UAECs, but not in NP-UAECs. Pretreatment with combined 1 mmol/L 1,3-bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole dihydrochloride (MPP; ERa antagonist) and 1 mmol/L 4-[2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-3-yl]phenol (PHTPP; ER-b antagonist) inhibited the rises in NOx levels stimulated by E 2 b and 2-ME 2 , but had no effect on 2-OHE 2-, 4-OHE 2-, or 4-ME 2-stimulated rises in NOx levels. Pretreatment with yohimbine (a 2-AR antagonist) and propranolol (b 2,3-AR antagonist) inhibited the rises in NOx levels stimulated by 2-OHE 2 , but not by E 2 b, 4-OHE 2 , 2-ME 2 , or 4-ME 2. These data demonstrate that E 2 b metabolites stimulate NO synthesis via ERs or ARs in UAECs in a pregnancy-specific manner, suggesting that these metabolites contribute to rises in vasodilation and UBF during pregnancy.

Biology of Reproduction, 2010

Intrauterine growth restriction (IUGR) is observed in conditions with limitations in uterine spac... more Intrauterine growth restriction (IUGR) is observed in conditions with limitations in uterine space (e.g., uterine anomalies and multifetal gestations). IUGR is associated with reduced fetal weight, organ growth, and a spectrum of adult-onset diseases. To examine the interaction of uterine anomalies and multifetal gestations, we developed a surgical uterine space restriction model with a unilateral uterine horn ligation before breeding (unilateral surgery). Placentas and fetuses were studied on Gestational Day (GD) 120 and GD 130 (term ¼ 147 days). Unilateral surgery decreased placentome numbers in singleton and twin pregnancies (25% and 50%, respectively) but not unilateral triplets. Unilateral surgery decreased total placentome weight in twin pregnancies (decreased 24%). Fetuses categorized as uterine space restricted (unilateral twin and both groups of triplets) had 51% fewer placentomes per fetus and a 31% reduction in placentomal weight per fetus compared to the nonrestricted group (control singleton, unilateral singleton, and control twin). By GD 130, uterine space-restricted fetuses exhibited decreased weight, smaller crown-rump, abdominal girth, and thoracic girth as well as decreased fetal heart, kidney, liver, spleen, and thymus weights. Lung and brain weights were unaffected, demonstrating asymmetric IUGR. At GD 130, placental efficiency (fetal weight per total placentomal weight) was elevated in uterine space-restricted fetuses. However, fetal arterial creatinine, blood urea nitrogen, and cholesterol were elevated, suggesting insufficient placental clearance. Maternalto-fetal glucose and triglycerides ratios were elevated in the uterine space-restricted pregnancies, suggesting placental nutrient transport insufficiency. This model allows for examination of interactive effects of uterine space restriction-induced IUGR on placental adaptation and fetal organ growth.

Seminars in Perinatology, 1997

Endocrinology, 2004

Rapid uterine vasodilatation after estrogen administration is believed to be mediated by endothel... more Rapid uterine vasodilatation after estrogen administration is believed to be mediated by endothelial production of nitric oxide (NO) via endothelial NO synthase (eNOS). However, the mechanism(s) by which estrogen activates eNOS in uterine artery endothelial cells (UAEC) is unknown. In this study, we observed that estradiol-17β (E2) and E2-BSA rapidly (<2 min) increased total NOx production in UAEC in vitro. This was associated with rapid eNOS phosphorylation and activation but was unaltered by pretreatment with actinomycin-D. estrogen receptor-α protein was detectable in isolated plasma membrane proteins by immunoblotting, and E2-BSA-fluorescein isothiocyanate binding was evident on the plasma membrane of UAEC. E2 did not mobilize intracellular Ca2+, but E2 and ionomycin in combination induced greater eNOS phosphorylation than either E2 or ionomycin alone. E2 did not stimulate rapid Akt phosphorylation. E2 stimulated rapid ERK2/1 activation in a time- and dose-dependent manner, w...

Alcoholism: Clinical and Experimental Research, 2011

Background-Pregnancy-induced utero-placental growth, angiogenic remodeling, and enhanced vasodila... more Background-Pregnancy-induced utero-placental growth, angiogenic remodeling, and enhanced vasodilation are all partly regulated by estradiol-17β-mediated activation of endothelial nitric oxide synthase (eNOS) and nitric oxide (NO) production. However, very little is known about the effects of alcohol on these maternal utero-placental vascular adaptations during pregnancy and its potential role in the pathogenesis of Fetal Alcohol Spectrum Disorders (FASD). In this study, we hypothesized that in vitro chronic binge-like alcohol will decrease uterine arterial endothelial eNOS expression and alter its multi-site phosphorylation activity state via disruption of AKT signaling. To study the direct effects of alcohol on uterine vascular adaptations, we further investigated the effects of alcohol on estradiol-17β-induced uterine angiogenesis in vitro. Methods-Uterine artery endothelial cells were isolated from pregnant ewes (gestational day 120-130; term = 147), Fluorescence Activated Cell sorted, validated, and maintained in culture to passage 4. To mimic maternal binge drinking patterns, cells were cultured in the absence or presence of a lower (LD) or higher dose (HD) of alcohol in a compensating sealed humidified chamber system equilibrated with aqueous alcohol for 3 h on 3 consecutive days. Immunoblotting was performed to assess expression of NO system-associated proteins and eNOS multi-site phosphorylation. Following this treatment paradigm, control and binge alcohol treated cells were passaged, grown for two days, and then treated with increasing concentrations of estradiol-17β (0.1, 1, 10, 100 nM) in the absence or presence of LD or HD alcohol to evaluate estradiol-17βinduced angiogenesis index using BrdU Proliferation Assay. Results-LD and HD binge-like alcohol decreased uterine arterial eNOS expression (P=0.009). eNOS multi-site phosphorylation activation state was altered: P 635 eNOS was decreased (P=0.017), P 1177 eNOS was not altered, and P 495 eNOS exhibited an inverse U shaped dose-dependent relationship with alcohol. LD and HD alcohol decreased the major eNOS-associated protein cav-1 (P<0.

24th Joint Propulsion Conference, 1988

Abstract—ATP leads to endothelial NO synthase (eNOS)/NO-mediated vasodilation, a process hypothes... more Abstract—ATP leads to endothelial NO synthase (eNOS)/NO-mediated vasodilation, a process hypothesized to depend on the endothelial caveolar eNOS partitioning and subcellular domain-specific multisite phosphorylation state. We demonstrate herein that, in both the absence and presence of ATP, the uterine artery endothelial caveolae contain specific protein machinery related to subcellular partitioning and act as specific focal “hubs ” for NO- and ATP-related proteins. ATP-induced eNOS regulation showed a complex set of multisite posttranslational phosphorylation events that were closely associated with the enzyme’s partitioning between caveolar and noncaveolar endothelial subcellular domains. The comprehensive model that we present demonstrates that ATP repartitioned eNOS between the caveolar and noncaveolar subcellular domains; specifically, the stimulatory PSer635eNOS was substantially higher in the caveolar pool with subcellular domain-independent increased levels on ATP treatment....

Biology of Reproduction, 2016

Endothelial nitric oxide (NO) production is partly responsible for maintenance of uterine vasodil... more Endothelial nitric oxide (NO) production is partly responsible for maintenance of uterine vasodilatation during physiologic states of high circulating estrogen levels, e.g., pregnancy. Although 3%-5% of estrogen receptors (ER-alpha/beta) localize to the endothelial plasmalemma, these receptors are responsible for the nongenomic vasodilator responses. Estradiol induces endothelial NO synthase (eNOS) activation to increase NO production; however, it is unknown if eNOS regulation is dependent on both ERs. We hypothesize that ER-alpha and/or ER-beta are capable of changing eNOS phosphorylation and increasing NO production in uterine artery endothelial cells (UAECs). UAECs were 1) treated with vehicle or increasing concentrations (0.1-100 nM) or timed treatments (0-30 min) of estradiol and 2) pretreated with the inhibitors ICI 182,780 (nonspecific ER), 1,3-Bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole dihydrochloride (MPP; ER-alpha specific), or 4-[2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-3-yl]phenol (PHTPP; ER-beta specific) followed by estradiol to analyze the changes in eNOS stimulatory Ser1177 eNOS and Ser635 eNOS versus inhibitory Thr495 e-NOS via Western blot analysis. UAECs were also pretreated with MPP, PHTPP, or MPP + PHTTP followed by estradiol or treated with the agonists estradiol, 4,4 0 ,4 00-(4-propyl-[1H]-pyrazole-1,3,5triyl)trisphenol, 2,3-bis(4-hydroxyphenyl)-propionitrile, or ATP to quantify total NOx levels (NO 2 +NO 3). Estrogen and ER-alpha activation induced an increase in Ser1177 eNOS and Ser635 eNOS, a decrease in Thr495 eNOS, and an increase in NOx levels. In contrast, ER-beta activation only reduced Thr495 eNOS without changes in Ser1177 eNOS or Ser635 eNOS. However, ER-beta activation increased NOx levels. Lastly, the antagonism of both receptors induced a reduction in basal and stimulated NOx levels in UAECs. These data demonstrate that 1) eNOS phosphorylation changes occur via ER-alpha-and ER-beta-dependent mechanisms and 2) ER-alpha and ER-beta can both increase NO levels independently from each other.

Trophoblast cell lines are important research tools used as a surrogate for primary trophoblast c... more Trophoblast cell lines are important research tools used as a surrogate for primary trophoblast cells in the study of placental function. Because the cellular origins of transformed trophoblasts are likely to be diverse, it would be of value to understand the unique and shared phenotypes of the cells on a global scale. We have compared two widely used cell lines, BeWo and JEG3, by microarray analysis in order to identify differentially expressed genes. Results indicated that approximately 2700 genes were differentially expressed between the cell lines, with principal differences observed in the biological processes of response to stress, cell adhesion, signal transduction, and protein and nucleobase metabolisms. These data suggest that BeWo and JEG3 cell lines, and perhaps other trophoblast cell lines, are sufficiently dissimilar from each other such that they will be differentially suited for specific experimental paradigms.

Objective: To evaluate vascular dysfunction using both physiologic measures and biochemical marke... more Objective: To evaluate vascular dysfunction using both physiologic measures and biochemical markers, longitudinally, prior to and during pregnancy, in nulliparous women who had uncomplicated pregnancies compared to those who developed complicated hypertension during pregnancy. Methods: Twenty healthy nulliparous women were studied during the follicular phase and in early (EP) and late (LP) pregnancy. All had singleton conceptions and delivered at term, seventeen with uncomplicated pregnancies (NP) and three who developed complicated hypertension (HP) after the LP evaluation. We compared prepregnancy, EP and LP pulse wave velocity (PWV) and soluble vascular cell adhesion molecule (sVCAM-1) between the NP and HP groups. PWV was measured using ultrasound and simultaneous echocardiogram tracing then calculated as the estimated distance divided by the interval between EKG R-wave peak and peak brachial artery flow. SVCAM-1 was measured using a commercially available kit. Data are mean ± SE, significance accepted as p < 0.05. Results: The NP group had significantly lower prepregnant PWV

American Journal of …, 1998

Prolonged 17 -estradiol (E 2 ) infusion decreases mean arterial pressure (MAP) and systemic vascu... more Prolonged 17 -estradiol (E 2 ) infusion decreases mean arterial pressure (MAP) and systemic vascular resistance (SVR) while increasing heart rate (HR) and cardiac output (CO). It is unclear, however, which systemic vascular beds show increases in perfusion. The ...

Experimental Brain Research, 1984

Lamina IV stellate cells and lamina V pyramidal cells were studied in Golgi material of visual co... more Lamina IV stellate cells and lamina V pyramidal cells were studied in Golgi material of visual cortex of rabbits ranging in age from 10 days to adult. Spine density counts revealed that primary branches have lower spine density than secondary or tertiary branches in both stellate and basilar pyramidal dendrites. Most areas sampled showed an increase in spine density from age 10 days to a peak at 25-30 days. In some areas this was followed by a plateau, but in most dendritic areas sampled there was a significant decrease from peak levels to adult levels. Measurements of dendritic length revealed that basilar dendrites undergo changes in length which parallel the changes in spine density counts: a peak in the length of basilar dendrites was followed by a decline to adult levels. However, the dendritic length of stellate cells showed much less change with age after 10 days. We propose that the time period during which spine density and pyramidal cell dendritic organization peaks above adult levels may coincide with, and provide a morphological correlate of, the critical period.

Journal of the Society for Gynecologic Investigation