scholarly journals Hypercholesterolemia impairs oxytocin-induced uterine contractility in late pregnant mouse

Reproduction ◽  
2017 ◽  
Vol 153 (5) ◽  
pp. 565-576 ◽  
Author(s):  
Amol R Padol ◽  
Susanth V Sukumaran ◽  
Abdul Sadam ◽  
Manickam Kesavan ◽  
Kandasamy Arunvikram ◽  
...  
Keyword(s):  
Pasteurella Multocida ◽  
Ex Vivo ◽  
Late Pregnancy ◽  
Sodium Cholate ◽  
Pregnant Mouse ◽  
Standard Diet ◽  
Uterine Contractility ◽  
High Cholesterol ◽  
Mouse Uterus

High cholesterol is known to negatively affect uterine contractility inex vivoconditions. The aim of the present study was to reveal the effect ofin vivohypercholesterolemia on spontaneous and oxytocin-induced uterine contractility in late pregnant mouse uterus. Female Swiss albino mice were fed with high cholesterol (HC) diet (0.5% sodium cholate, 1.25% cholesterol and 15% fat) for 6 weeks and then throughout the gestation period after mating. On day 19 of gestation, serum cholesterol level was increased more than 3-fold while triglycerides level was reduced in HC diet-fed animals as compared to control animals fed with a standard diet. In tension experiments, neither the mean integral tension of spontaneous contractility nor the response to CaCl2in high K+-depolarized tissues was altered, but the oxytocin-induced concentration-dependent contractile response in uterine strips was attenuated in hypercholesterolemic mice as compared to control. Similarly, hypercholesterolemia dampened concentration-dependent uterine contractions elicited by a GNAQ protein activator,Pasteurella multocidatoxin. However, it had no effect on endogenous oxytocin level either in plasma or in uterine tissue. It also did not affect the prostaglandin release in oxytocin-stimulated tissues. Western blot data showed a significant increase in caveolin-1 and GRK6 proteins but decline in oxytocin receptor, GNAQ and RHOA protein expressions in hypercholesterolemic mouse uterus. The results of the present study suggest that hypercholesterolemia may attenuate the uterotonic action of oxytocin in late pregnancy by causing downregulation of oxytocin receptors and suppressing the signaling efficacy through GNAQ and RHOA proteins.

scholarly journals Standardized Aronia melanocarpa Extract as Novel Supplement against Metabolic Syndrome: A Rat Model

2018 ◽  
Vol 20 (1) ◽  
pp. 6 ◽  
Author(s):  
Vladimir Jakoviljevic ◽  
Petar Milic ◽  
Jovana Bradic ◽  
Jovana Jeremic ◽  
Vladimir Zivkovic ◽  
...  
Keyword(s):  
Metabolic Syndrome ◽  
Dietary Habits ◽  
Ex Vivo ◽  
Heart Function ◽  
Standard Diet ◽  
Oral Supplementation ◽  
Beneficial Effects ◽  
Aronia Melanocarpa ◽  
Dietary Strategies

The aim of our study was to examine the effects of different dietary strategies, high-fat (HFd) or standard diet (Sd) alone or in combination with standardized oral supplementation (0.45 mL/kg/day) of Aronia melanocarpa extract (SAE) in rats with metabolic syndrome (MetS). SAE is an official product of pharmaceutical company Pharmanova (Belgrade, Serbia); however, the procedure for extraction was done by EU-Chem company (Belgrade, Serbia). Rats were divided randomly into six groups: control with Sd, control with Sd and SAE, MetS with HFd, MetS with HFd and SAE, MetS with Sd and MetS with Sd and SAE during 4 weeks. At the end of the 4-week protocol, cardiac function and liver morphology were assessed, while in the blood samples glucose, insulin, iron levels and systemic redox state were determined. Our results demonstrated that SAE had the ability to lower blood pressure and exert benefits on in vivo and ex vivo heart function. Moreover, SAE improved glucose tolerance, attenuated pathological liver alterations and oxidative stress present in MetS. Obtained beneficial effects of SAE were more prominent in combination with changing dietary habits. Promising potential of SAE supplementation alone or in combination with different dietary protocols in triggering cardioprotection should be further examined in future.

scholarly journals Progesterone receptor isoform B regulates the Oxtr-Plcl2-Trpc3 pathway to suppress uterine contractility

2021 ◽  
Vol 118 (11) ◽  
pp. e2011643118
Author(s):  
Mary C. Peavey ◽  
San-Pin Wu ◽  
Rong Li ◽  
Jian Liu ◽  
Olivia M. Emery ◽  
...  
Keyword(s):  
Progesterone Receptor ◽  
Ex Vivo ◽  
Muscle Relaxation ◽  
Gene Signature ◽  
Genetic Profile ◽  
Uterine Contractility ◽  
Gestational Length ◽  
In Vivo Experiments ◽  
Labor Dystocia

Uterine contractile dysfunction leads to pregnancy complications such as preterm birth and labor dystocia. In humans, it is hypothesized that progesterone receptor isoform PGR-B promotes a relaxed state of the myometrium, and PGR-A facilitates uterine contraction. This hypothesis was tested in vivo using transgenic mouse models that overexpress PGR-A or PGR-B in smooth muscle cells. Elevated PGR-B abundance results in a marked increase in gestational length compared to control mice (21.1 versus 19.1 d respectively, P < 0.05). In both ex vivo and in vivo experiments, PGR-B overexpression leads to prolonged labor, a significant decrease in uterine contractility, and a high incidence of labor dystocia. Conversely, PGR-A overexpression leads to an increase in uterine contractility without a change in gestational length. Uterine RNA sequencing at midpregnancy identified 1,174 isoform-specific downstream targets and 424 genes that are commonly regulated by both PGR isoforms. Gene signature analyses further reveal PGR-B for muscle relaxation and PGR-A being proinflammatory. Elevated PGR-B abundance reduces Oxtr and Trpc3 and increases Plcl2 expression, which manifests a genetic profile of compromised oxytocin signaling. Functionally, both endogenous PLCL2 and its paralog PLCL1 can attenuate uterine muscle cell contraction in a CRISPRa-based assay system. These findings provide in vivo support that PGR isoform levels determine distinct transcriptomic landscapes and pathways in myometrial function and labor, which may help further the understanding of abnormal uterine function in the clinical setting.

scholarly journals Dickkopf-1 secreted by decidual cells promotes trophoblast cell invasion during murine placentation

Reproduction ◽  
2008 ◽  
Vol 135 (3) ◽  
pp. 367-375 ◽  
Author(s):  
Sha Peng ◽  
Jing Li ◽  
Chenglin Miao ◽  
Liwei Jia ◽  
Zeng Hu ◽  
...  
Keyword(s):  
Cell Invasion ◽  
Wnt Signaling Pathway ◽  
Pregnant Mouse ◽  
Trophoblast Cell ◽  
Mouse Uterus ◽  
Recombinant Mouse ◽  
Decidual Cells ◽  
Dickkopf 1

Dickkopf-1 (Dkk1) is one of the secreted antagonists in the canonical Wnt signaling pathway. It plays important roles in diverse developmental processes. However, the role of Dkk1 in trophoblast cell invasion during placentation remains unclear. In this study, we found that Dkk1 was mainly expressed in maternal decidual tissue but trivially in ectoplacental cones (EPCs) in day 8 post coitum (p.c.) pregnant mouse uterus and that the efficiency of EPC attachment and outgrowth was increased when co-cultured with decidual cells, which secreted Dkk1, and this enhancement was abolished by pretreating decidual cells with Dkk1 blocking antibody before co-culture experiment. This indicates that Dkk1 secreted by decidual cells plays an important role in trophoblast cell invasion. Indeed, when recombinant mouse Dkk1 was added to EPCs in vitro, the efficiency of attachment and outgrowth was increased. Migration of EPCs toward the decidua was retarded when antisense Dkk1 oligonucleotide (ODN) was administered via intrauterine injection in vivo. Furthermore, the active β-catenin nuclear location was lost when we treated cultured EPCs with recombinant mouse Dkk1, and the efficiency of EPCs attachment and outgrowth was obviously increased when we treated cultured EPCs with antisense β-catenin ODN. Taken together, Dkk1 secreted by decidual cells may induce trophoblast cell invasion in the mouse and β-catenin may be involved in such functions of Dkk1.

Simple and highly efficient method for transient in vivo gene transfer to mid-late pregnant mouse uterus

2006 ◽  
Vol 70 (1-2) ◽  
pp. 59-69 ◽  
Author(s):  
Shinsuke Koyama ◽  
Tadashi Kimura ◽  
Kazuhide Ogita ◽  
Hitomi Nakamura ◽  
Chisa Tabata ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Efficient Method ◽  
Pregnant Mouse ◽  
Mouse Uterus ◽  
Highly Efficient ◽  
In Vivo Gene Transfer

scholarly journals Modelling maternal obesity: the effects of a chronic high-fat, high-cholesterol diet on uterine expression of contractile-associated proteins and ex vivo contractile activity during labour in the rat

2015 ◽  
Vol 130 (3) ◽  
pp. 183-192 ◽  
Author(s):  
Ronan Muir ◽  
Jean Ballan ◽  
Bethan Clifford ◽  
Sarah McMullen ◽  
Raheela Khan ◽  
...  
Keyword(s):  
Maternal Obesity ◽  
Ex Vivo ◽  
Contractile Activity ◽  
High Cholesterol Diet ◽  
Obese Women ◽  
Uterine Contractility ◽  
High Cholesterol ◽  
Steroid Synthesis ◽  
Uterine Contractile ◽  
Associated Proteins

Modelling maternal obesity in rats adversely affected steroid synthesis, uterine contractile associated protein expression and ex-vivo uterine contractility during labour. This maternal obesity model can be utilized further to unravel the mechanisms causing uterine dystocia in obese women.

scholarly journals P0020THE NEPHRO-PROTECTIVE ROUTE OF CGMP DOES NOT COMPENSATE NPRHOGENIC DIABETES INSIPIDUS, BOTH CONDITIONS MEDIATED BY RENAL INTEGRIN LINKED KINASE (ILK)

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Mercedes Griera-Merino ◽  
Diego García-Ayuso ◽  
Elena Gutiérrez-Calabrés ◽  
Lourdes Bohorquez Magro ◽  
Sofía Campillo de Blas ◽  
...  
Keyword(s):  
Diabetes Insipidus ◽  
Apical Membrane ◽  
Ex Vivo ◽  
Water Channel ◽  
Urine Volume ◽  
Renal Diseases ◽  
Standard Diet ◽  
No Donor ◽  
Integrin Linked Kinase

Abstract Background and Aims Renal ILK transgenic deletion on mice (cKD-ILK) address basal polyuria compatible with nephrogenic diabetes insipidus (NDI), due to disrupted tubular water channel aquaporin 2 (AQP2) AQP2 expression and vesicular trafficking to the apical membrane [Cano-Peñalver JL et al. FASEB J. 2014; Mamuya FA et al. Am J Physiol Renal Physiol. 2016]. An early symptom of chronic renal diseases (CKD) is NDI. ILK depletion also have a nephron-protective effect in renal damage in mice due to the increase in cGMP [Cano-Peñalver JL et al. Mol Med. 2016; de Frutos S et al. Biochim Biophys Acta Mol Basis Dis. 2019], the latter an alternative anti-NDI mediator. Here we demonstrate that ILK-mediated regulation of AQP2, under basal or CKD contexts, does not depend on cGMP. Method cKD-ILK and controls (WT) were treated orally with a NO donor, as precursor of cGMP formation (IDN, 300mg / Kg / day) for 24 hours. Urine volume was determined as a measure of tubular functionality. Fresh kidneys from non-treated mice were cultured ex vivo with precursors of cGMP (NO donor SNP, 1 µM or cGMP analogue 8Br-cGMP, 0.2 mM) for 30 minutes. AQP2 in the apical membrane of the tubules was quantified by immunohistochemistry. More cKD-ILK and WT were subjected for 6 weeks to a diet supplemented with 0.2% adenine (A) as CKD inducer or baseline standard diet. Urine volume and total ranal AQP2 levels were determined. Results Compared to WT, cGMP axis activation in vivo did not compensate the basal NDI present in cKD-ILK and the activation ex vivo did not improve the apical presence of AQP2 in cKD-ILK. Both WT and cKD-ILK subjected to CKD showed exacerbated polyuria and decreased levels of renal AQP2, but no differences were observed between groups. Conclusion ILK regulates AQP2, both in a basal and pathological context (CKD) independent of cGMP which otherwise is nephron-protective. Our results probably explain the co-existence of cGMP increase together with AQP2 decrease observed in the cKD-ILK kidneys.

scholarly journals Monitoring uterine contractility in mice using a transcervical intrauterine pressure catheter

Reproduction ◽  
2018 ◽  
Vol 155 (5) ◽  
pp. 447-456 ◽  
Author(s):  
Michael F Robuck ◽  
Christine M O’Brien ◽  
Kelsi M Knapp ◽  
Sheila D Shay ◽  
James D West ◽  
...  
Keyword(s):  
Mouse Models ◽  
Ex Vivo ◽  
Contractile Activity ◽  
Area Under The Curve ◽  
Fetal Outcome ◽  
Isometric Tension ◽  
Uterine Contractility ◽  
Intrauterine Pressure ◽  
Pressure Catheter

In mouse models used to study parturition or pre-clinical therapeutic testing, measurement of uterine contractions is limited to either ex vivo isometric tension or operative intrauterine pressure (IUP). The goal of this study was to: (1) develop a method for transcervical insertion of a pressure catheter to measure in vivo intrauterine contractile pressure during mouse pregnancy, (2) determine whether this method can be utilized numerous times in a single mouse pregnancy without affecting the timing of delivery or fetal outcome and (3) compare the in vivo contractile activity between mouse models of term and preterm labor (PTL). Visualization of the cervix allowed intrauterine pressure catheter (IUPC) placement into anesthetized pregnant mice (plug = day 1, delivery = day 19.5). The amplitude, frequency, duration and area under the curve (AUC) of IUP was lowest on days 16–18, increased significantly (P < 0.05) on the morning of day 19 and reached maximal levels during by the afternoon of day 19 and into the intrapartum period. An AUC threshold of 2.77 mmHg discriminated between inactive labor (day 19 am) and active labor (day 19 pm and intrapartum period). Mice examined on a single vs every experimental timepoint did not have significantly different IUP, timing of delivery, offspring number or fetal/neonatal weight. The IUP was significantly greater in LPS-treated and RU486-treated mouse models of PTL compared to time-matched vehicle control mice. Intrapartum IUP was not significantly different between term and preterm mice. We conclude that utilization of a transcervical IUPC allows sensitive assessment of in vivo uterine contractile activity and labor progression in mouse models without the need for operative approaches.

scholarly journals A cDNA cloned from pregnant mouse uterus exhibits temporo-spatial expression and predicts a novel protein

1998 ◽  
Vol 330 (2) ◽  
pp. 947-950 ◽  
Author(s):  
W. John KASIK
Keyword(s):  
Late Pregnancy ◽  
Pregnant Mouse ◽  
Open Reading Frame ◽  
Mouse Uterus ◽  
Pregnant Uterus ◽  
Reading Frame ◽  
Spatial Expression ◽  
Conceptual Translation ◽  
Novel Protein ◽  
Specific Mrna

A cDNA was cloned from a pregnant mouse uterus cDNA library. On conceptual translation, the cDNA has one long open reading frame that predicts a novel protein of 606 amino acids. This protein is principally composed of two CUB domains and a ZP domain; motifs found in proteins implicated in egg-sperm recognition. Probes derived from the cDNA were used to conduct Northern hybridizations. The expression of this mRNA is temporal; message first appears in the uterus 6 days prior to birth, it increases each subsequent day to attain maximal levels at 3 days prior to birth and then abruptly decreases during the last 3 days of pregnancy. The expression of this mRNA is restricted; message is abundant in the uterus during late pregnancy, but it is not found in non-pregnant uterus or in a variety of adult or fetal tissues. The temporo-spatial expression of this pregnant uterus specific mRNA and the consolidation in the predicted protein of two motifs implicated in early pregnancy events suggests that the product of the gene represented by this mRNA may play an important role in events that transpire during late pregnancy.

scholarly journals OR20-05 Regulation of Mouse Uterine Contractility by Regulator of G-Protein Signalling 2

2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Daniela Urrego ◽  
Stephen L Wood ◽  
Robert Newton ◽  
Donna Michelle Slater
Keyword(s):  
G Protein ◽  
Preterm Labour ◽  
Ex Vivo ◽  
Isometric Force ◽  
Pregnant Mouse ◽  
Receptor Expression ◽  
Human Myometrium ◽  
Protein Signaling ◽  
Uterine Contractility ◽  
Student’S T

Abstract INTRODUCTION: Uterine contractions in labour are produced by activation of receptors such as FP (PGF2α) and OXTR (oxytocin). Such receptors signal via Gαq proteins that can be selectively turned off by the regulator of G-protein signaling 2 (RGS2). RGS2 expression can be upregulated by Gαs-coupled receptor signaling. PGE2 elicits various uterine effects, some via activation of Gαs-coupled receptors. In primary human myometrial cells, PGE2 enhances RGS2 expression that attenuates subsequent oxytocin-stimulated calcium responses associated with contraction. RGS2 expression is higher in preterm non-labour versus labour human myometrium, suggesting that RGS2 promotes quiescence in the pregnant uterus. To directly study whether RGS2 dampens uterine contraction, we analyzed the ex vivo contractility of pregnant mouse uteri. We hypothesize that loss of RGS2 expression enhances oxytocin and PGF2α-stimulated uterine contractility. METHODS: Wildtype (WT) and RGS2 knockout (KO) female mice (8-14 weeks) were time-mated and euthanized on days 15-20 of pregnancy or during active labour (n=5-11 per group). Uteri were snap frozen for RNA/protein expression analysis by qPCR and western blot. Uterine sections from d18-19 were dissected into longitudinal strips for ex vivo contractility measurement. Tissues were submerged in Krebs-Henseleit buffer and tied to force transducers to measure isometric force development. Tissues were treated with or without 1μM PGE2 for 2 hours. In parallel, pretreated and non-pretreated tissues were exposed to increasing concentrations of oxytocin or PGF2α (0.1-10uM). All tissues were treated with 100mM KCl to determine the maximum contraction at the end of the experiment. Data were analyzed by ANOVA (Bonferroni post-hoc), or Student’s t-test, as appropriate. RESULTS: RGS2 expression in WT uteri sharply decreases at day 19 (p&lt;0.05), remaining low during labour. OXTR and FP receptor expression increase at day 19 (p&lt;0.01 and p&lt;0.05, respectively) and are sustained through labour. Compared to WT, uteri from KO mice produced larger oxytocin-stimulated contractile amplitudes (p&lt;0.05), and more frequent PGF2α-stimulated contractions (p&lt;0.05). PGE2 pretreatment enhanced RGS2 expression in WT uteri, which reduced the peak amplitudes and integrals elicited by subsequent oxytocin treatment, compared to non-pretreated uteri (p&lt;0.05). CONCLUSION: RGS2 may play a pro-quiescent role in pregnancy, since RGS2 loss enhances uterine contractility. Concomitant upregulation of pro-contractile receptors and RGS2 downregulation may facilitate labour activation. This mechanisms may be involved in the etiology of spontaneous preterm labour; loss of RGS2 expression observed in human preterm labour may allow for early contractile activation. Understanding RGS2’s role in quiescence is important to developing more effective strategies for managing preterm labour.

The Role of Polyphenols in the Modulation of Plasma Non-Enzymatic Antioxidant Capacity (NEAC)

2012 ◽  
Vol 82 (3) ◽  
pp. 228-232 ◽  
Author(s):  
Mauro Serafini ◽  
Giuseppa Morabito
Keyword(s):  
Antioxidant Capacity ◽  
Dietary Intervention ◽  
Ex Vivo ◽  
The Body ◽  
Dietary Polyphenols ◽  
Enzymatic Antioxidant ◽  
Endogenous Antioxidant

Dietary polyphenols have been shown to scavenge free radicals, modulating cellular redox transcription factors in different in vitro and ex vivo models. Dietary intervention studies have shown that consumption of plant foods modulates plasma Non-Enzymatic Antioxidant Capacity (NEAC), a biomarker of the endogenous antioxidant network, in human subjects. However, the identification of the molecules responsible for this effect are yet to be obtained and evidences of an antioxidant in vivo action of polyphenols are conflicting. There is a clear discrepancy between polyphenols (PP) concentration in body fluids and the extent of increase of plasma NEAC. The low degree of absorption and the extensive metabolism of PP within the body have raised questions about their contribution to the endogenous antioxidant network. This work will discuss the role of polyphenols from galenic preparation, food extracts, and selected dietary sources as modulators of plasma NEAC in humans.

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