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Article: Mapping uterine calcium dynamics during the ovulatory cycle in live mice.

Combs, David J / Moult, Eric M / England, Sarah K / Cohen, Adam E

bioRxiv : the preprint server for biology

2024

Abstract: Uterine contraction patterns vary during the ovulatory cycle and throughout pregnancy but prior measurements have produced limited and conflicting information on these patterns. We combined a virally delivered genetically encoded calcium reporter ( ... ...

Abstract	Uterine contraction patterns vary during the ovulatory cycle and throughout pregnancy but prior measurements have produced limited and conflicting information on these patterns. We combined a virally delivered genetically encoded calcium reporter (GCaMP8m) and ultra-widefield imaging in live nonpregnant mice to characterize uterine calcium dynamics at organ scale throughout the estrous cycle. Prior to ovulation (proestrus and estrus) uterine excitations primarily initiated in a region near the oviduct, but after ovulation (metestrus and diestrus), excitations initiated at loci homogeneously distributed throughout the organ. The frequency of excitation events was lowest in proestrus and estrus, higher in metestrus and highest in diestrus. These results establish a platform for mapping uterine activity, and show that the question of whether there is an anatomically localized trigger for uterine excitations depends on the estrous cycle phase.
Language	English
Publishing date	2024-04-06
Publishing country	United States
Document type	Preprint
DOI	10.1101/2024.02.02.578395
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Article ; Online: Solving the Puzzle of Preterm Birth.

Stevenson, David K / Winn, Virginia D / Shaw, Gary M / England, Sarah K / Wong, Ronald J

Clinics in perinatology

2024 Volume 51, Issue 2, Page(s) 291–300

Abstract: Solving the puzzle of preterm birth has been challenging and will require novel integrative solutions as preterm birth likely arises from many etiologies. It has been demonstrated that many sociodemographic and psychological determinants of preterm birth ...

Abstract	Solving the puzzle of preterm birth has been challenging and will require novel integrative solutions as preterm birth likely arises from many etiologies. It has been demonstrated that many sociodemographic and psychological determinants of preterm birth relate to its complex biology. It is this understanding that has enabled the development of a novel preventative strategy, which integrates the omics profile (genome, epigenome, transcriptome, proteome, metabolome, microbiome) with sociodemographic, environmental, and psychological determinants of individual pregnant people to solve the puzzle of preterm birth.
MeSH term(s)	Humans ; Premature Birth/epidemiology ; Female ; Pregnancy ; Infant, Newborn ; Risk Factors
Language	English
Publishing date	2024-03-08
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	193116-7
ISSN	1557-9840 ; 0095-5108
ISSN (online)	1557-9840
ISSN	0095-5108
DOI	10.1016/j.clp.2024.02.001
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Article ; Online: Absolute Quantification of Plasma Membrane Receptors Via Quantitative Flow Cytometry.

Fang, Yingye / Malik, Manasi / England, Sarah K / Imoukhuede, P I

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2475, Page(s) 61–77

Abstract: Plasma membrane receptors are transmembrane proteins that initiate cellular response following the binding of specific ligands (e.g., growth factors, hormones, and cytokines). The abundance of plasma membrane receptors can be a diagnostic or prognostic ... ...

Abstract	Plasma membrane receptors are transmembrane proteins that initiate cellular response following the binding of specific ligands (e.g., growth factors, hormones, and cytokines). The abundance of plasma membrane receptors can be a diagnostic or prognostic biomarker in many human diseases. One of the best techniques for measuring plasma membrane receptors is quantitative flow cytometry (qFlow). qFlow employs fluorophore-conjugated antibodies against the receptors of interest and corresponding fluorophore-loaded calibration beads offers standardized and reproducible measurements of plasma membrane receptors. More importantly, qFlow can achieve absolute quantification of plasma membrane receptors when phycoerythrin (PE) is the fluorophore of choice. Here we describe a detailed qFlow protocol to obtain absolute receptor quantities on the basis of PE calibration. This protocol is foundational for many previous and ongoing studies in quantifying tyrosine kinase receptors and G-protein-coupled receptors with in vitro cell models and ex vivo cell samples.
MeSH term(s)	Calibration ; Cell Membrane ; Flow Cytometry/methods ; Fluorescent Dyes ; Humans ; Phycoerythrin
Chemical Substances	Fluorescent Dyes ; Phycoerythrin (11016-17-4)
Language	English
Publishing date	2022-04-22
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, N.I.H., Extramural
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2217-9_4
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Article ; Online: Uterine contractions in rodent models and humans.

Malik, Manasi / Roh, Michelle / England, Sarah K

Acta physiologica (Oxford, England)

2021 Volume 231, Issue 4, Page(s) e13607

Abstract: Aberrant uterine contractions can lead to preterm birth and other labour complications and are a significant cause of maternal morbidity and mortality. To investigate the mechanisms underlying dysfunctional uterine contractions, researchers have used ... ...

Abstract	Aberrant uterine contractions can lead to preterm birth and other labour complications and are a significant cause of maternal morbidity and mortality. To investigate the mechanisms underlying dysfunctional uterine contractions, researchers have used experimentally tractable small animal models. However, biological differences between humans and rodents change how researchers select their animal model and interpret their results. Here, we provide a general review of studies of uterine excitation and contractions in mice, rats, guinea pigs, and humans, in an effort to introduce new researchers to the field and help in the design and interpretation of experiments in rodent models.
MeSH term(s)	Animals ; Female ; Guinea Pigs ; Humans ; Infant, Newborn ; Mice ; Myometrium ; Pregnancy ; Premature Birth ; Rats ; Rodentia ; Uterine Contraction ; Uterus
Language	English
Publishing date	2021-01-07
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2218636-0
ISSN	1748-1716 ; 1748-1708
ISSN (online)	1748-1716
ISSN	1748-1708
DOI	10.1111/apha.13607
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Article: Quantification of surface-localized and total oxytocin receptor in myometrial smooth muscle cells.

Fang, Yingye / Reinl, Erin L / Liu, Audrey / Prochaska, Trinidi D / Malik, Manasi / Frolova, Antonina I / England, Sarah K / Imoukhuede, Princess I

Heliyon

2024 Volume 10, Issue 4, Page(s) e25761

Abstract: Oxytocin acts through the oxytocin receptor (OXTR) to modulate uterine contractility. We previously identified OXTR genetic variants and showed that, in HEK293T cells, two of the OXTR protein variants localized to the cell surface less than wild-type ... ...

Abstract	Oxytocin acts through the oxytocin receptor (OXTR) to modulate uterine contractility. We previously identified OXTR genetic variants and showed that, in HEK293T cells, two of the OXTR protein variants localized to the cell surface less than wild-type OXTR. Here, we sought to measure OXTR in the more native human myometrial smooth muscle cell (HMSMC) line on both the cell-surface and across the whole cell, and used CRISPR editing to add an HA tag to the endogenous OXTR gene for anti-HA measurement. Quantitative flow cytometry revealed that these cells possessed 55,000 ± 3200 total OXTRs and 4900 ± 390 cell-surface OXTRs per cell. To identify any differential wild-type versus variant localization, we transiently transfected HMSMCs to exogenously express wild-type or variant OXTR with HA and green fluorescent protein tags. Total protein expression of wild-type OXTR and all tested variants were similar. However, the two variants with lower surface localization in HEK293T cells also presented lower surface localization in HMSMCs. Overall, we confirm the differential surface localization of variant OXTR in a more native cell type, and further demonstrate that the quantitative flow cytometry technique is adaptable to whole-cell measurements.
Language	English
Publishing date	2024-02-13
Publishing country	England
Document type	Journal Article
ZDB-ID	2835763-2
ISSN	2405-8440
ISSN	2405-8440
DOI	10.1016/j.heliyon.2024.e25761
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Article ; Online: Risk of pre-term birth as a function of sleep quality and obesity: prospective analysis in a large Prematurity Research Cohort.

Sutcliffe, Siobhan / Zhao, Peinan / Pilz, Luisa Klaus / Oakes, Megan / Frolova, Antonina I / Herzog, Erik D / England, Sarah K

Sleep advances : a journal of the Sleep Research Society

2023 Volume 4, Issue 1, Page(s) zpad043

Abstract: Study objective: To investigate whether poor sleep quality is associated with pre-term birth (PTB) risk, overall and independent of sleep apnea and habitual snoring.: Methods: We used longitudinal data from the Washington University Prematurity ... ...

Abstract	Study objective: To investigate whether poor sleep quality is associated with pre-term birth (PTB) risk, overall and independent of sleep apnea and habitual snoring. Methods: We used longitudinal data from the Washington University Prematurity Research Cohort to investigate the association between poor sleep quality (defined as a Pittsburgh Sleep Quality Index > 5) and PTB, overall and independent of sleep apnea and snoring (defined by the Berlin questionnaire and prior sleep clinic attendance). Associations were investigated for sleep quality early and throughout pregnancy. Stratified analyses were performed by factors previously shown to modify associations between sleep and PTB (race, pre-pregnancy obesity). Results: Of the 976 eligible participants, 50.1% experienced poor sleep quality early in pregnancy (<20 completed weeks) and 14.2% delivered pre-term ( Conclusion: Our findings suggest that improving sleep quality early in pregnancy may be important for PTB prevention, particularly among individuals with obesity.
Language	English
Publishing date	2023-11-02
Publishing country	United States
Document type	Journal Article
ISSN	2632-5012
ISSN (online)	2632-5012
DOI	10.1093/sleepadvances/zpad043
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Article ; Online: The Association between Mode of Transportation Support and Research Study Visit Attendance among Pregnant Patients.

Vinson, Ariel / Paul, Rachel / Chubiz, Jessica / Raghuraman, Nandini / Kelly, Jeannie C / England, Sarah K / Carter, Ebony B

American journal of perinatology

2023

Abstract: Objective: This study aimed to examine the association between transportation assistance and study visits, and explore differences by transportation modality.: Study design: This was a secondary analysis of prospective cohort study. We identified ... ...

Abstract	Objective: This study aimed to examine the association between transportation assistance and study visits, and explore differences by transportation modality. Study design: This was a secondary analysis of prospective cohort study. We identified patients requesting transportation support for research ultrasound visits and identified controls (1:2 ratio) who did not request support matched for age, race, and insurance type. Conditional logistic regression examined the association between transportation support and mode of transportation with study visit attendance. Results: Transportation support was requested by 57/1,184 (4.8%) participants. Participants that requested transportation support were three times more likely to attend visits than their matched controls (adjusted odds ratio [aOR] 3.16, 95% confidence interval [CI]: 1.76-5.68). Among visits with transportation support, those supported by a ridesharing service had five-fold higher odds of attendance than visits supported with taxi service (aOR 5.06, 95% CI: 1.50-16.98). Conclusion: Transportation support, especially a ridesharing service, is associated with improved attendance at research study visits in a sample of predominantly low-income, Black, pregnant participants. Implementing transportation support may be a promising strategy to improve engagement in research studies. Key points: · Participants utilizing transportation assistance were more likely to attend study appointments.. · Participants using ridesharing had higher likelihood of attendance than those using taxi service.. · Transportation assistance may improve research engagement for historically marginalized people..
Language	English
Publishing date	2023-12-15
Publishing country	United States
Document type	Journal Article
ZDB-ID	605671-4
ISSN	1098-8785 ; 0735-1631
ISSN (online)	1098-8785
ISSN	0735-1631
DOI	10.1055/s-0043-1777441
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Article ; Online: BKCa channels are involved in spontaneous and lipopolysaccharide-stimulated uterine contraction in late gestation mice†.

Bao, Junjie / Ma, Xiaofeng / Kent, Lindsey N / Wakle-Prabagaran, Monali / McCarthy, Ronald / England, Sarah K

Biology of reproduction

2023 Volume 110, Issue 4, Page(s) 798–807

Abstract: The large-conductance, voltage-gated, calcium (Ca2+)-activated potassium channel (BKCa) is one of the most abundant potassium channels in the myometrium. Previous work conducted by our group has identified a link between inflammation, BKCa channels and ... ...

Abstract	The large-conductance, voltage-gated, calcium (Ca2+)-activated potassium channel (BKCa) is one of the most abundant potassium channels in the myometrium. Previous work conducted by our group has identified a link between inflammation, BKCa channels and excitability of myometrial smooth muscle cells. Here, we investigate the role of BKCa channels in spontaneous and lipopolysaccharide (LPS)-stimulated uterine contraction to gain a better understanding of the relationship between the BKCa channel and uterine contraction in basal and inflammatory states. Uteri of C57BL/6 J mice on gestational day 18.5 (GD18.5) were obtained and either fixed in formalin or used immediately for tension recording or isolation of primary myocytes for patch-clamp. Paraffin sections were used for immunofluorescenctdetection of BKCa and Toll-like receptor (TLR4). For tension recordings, LPS was administered to determine its effect on uterine contractions. Paxilline, a BKCa inhibitor, was used to dissect the role of BKCa in uterine contraction in basal and inflammatory states. Finally, patch-clamp recordings were performed to investigate the relationship between LPS, the BKCa channel and membrane currents in mouse myometrial smooth muscle cells (mMSMCs). We confirmed the expression of BKCa and TLR4 in the myometrium of GD18.5 mice and found that inhibiting BKCa channels with paxilline suppressed both spontaneous and LPS-stimulated uterine contractions. Furthermore, application of BKCa inhibitors (paxilline or iberiotoxin) after LPS inhibited BKCa channel activity in mMSMCs. Moreover, pretreatment with BKCa inhibitor or the TLR4 inhibitor suppressed LPS-activated BKCa currents. Our study demonstrates that BKCa channels are involved in both basal and LPS-stimulated uterine contraction in pregnant mice.
MeSH term(s)	Animals ; Female ; Mice ; Pregnancy ; Large-Conductance Calcium-Activated Potassium Channels/metabolism ; Lipopolysaccharides/pharmacology ; Mice, Inbred C57BL ; Toll-Like Receptor 4/metabolism ; Uterine Contraction/drug effects ; Uterine Contraction/genetics ; Large-Conductance Calcium-Activated Potassium Channel alpha Subunits/metabolism
Chemical Substances	Large-Conductance Calcium-Activated Potassium Channels ; Lipopolysaccharides ; Toll-Like Receptor 4 ; BKCa protein, mouse ; Large-Conductance Calcium-Activated Potassium Channel alpha Subunits
Language	English
Publishing date	2023-12-23
Publishing country	United States
Document type	Journal Article
ZDB-ID	1118-6
ISSN	1529-7268 ; 0006-3363
ISSN (online)	1529-7268
ISSN	0006-3363
DOI	10.1093/biolre/ioad174
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Article: Understanding the effects of oxytocin receptor variants on OXT-OXT receptor binding: A mathematical model.

Dubey, Preeti / Fang, Yingye / Tukei, K Lionel / Kuila, Shobhan / Liu, Xinming / Sahota, Annika / Frolova, Antonina I / Reinl, Erin L / Malik, Manasi / England, Sarah K / Imoukhuede, Princess I

bioRxiv : the preprint server for biology

2024

Abstract: Approximately half of U.S. women giving birth annually receive Pitocin, the synthetic form of oxytocin (OXT), yet its effective dose can vary significantly. This variability presents safety concerns due to unpredictable responses, which may lead to ... ...

Abstract	Approximately half of U.S. women giving birth annually receive Pitocin, the synthetic form of oxytocin (OXT), yet its effective dose can vary significantly. This variability presents safety concerns due to unpredictable responses, which may lead to adverse outcomes for both mother and baby. To address the need for improved dosing, we developed a data-driven mathematical model to predict OXT receptor (OXTR) binding. Our study focuses on five prevalent OXTR variants (V45L, P108A, L206V, V281M, and E339K) and their impact on OXT-OXTR binding dynamics in two distinct cell types: human embryonic kidney cells (HEK293T), commonly used in experimental systems, and human myometrial smooth muscle cells, containing endogenous OXTR. We parameterized the model with cell-specific OXTR surface localization measurements. To strengthen the robustness of our study, we conducted a comprehensive meta-analysis of OXT- OXTR binding, enabling parameterization of our model with cell-specific OXT-OXTR binding kinetics (myometrial OXT-OXTR K
Language	English
Publishing date	2024-03-16
Publishing country	United States
Document type	Preprint
DOI	10.1101/2024.02.28.582600
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Article ; Online: Dietary risk factors for hypertensive disorders of pregnancy.

O'Nan, Suzanne L / Huang, Ruizhi / Zhao, Peinan / Garr Barry, Valene / Lawlor, Megan / Carter, Ebony B / Kelly, Jeannie C / Frolova, Antonina I / England, Sarah K / Raghuraman, Nandini

Pregnancy hypertension

2024 Volume 36, Page(s) 101120

Abstract: Objective: To assess whether diet quality and specific dietary components are associated with hypertensive disorders of pregnancy (HDP).: Study design: Nested case control study in a prospectively collected cohort of 450 participants with singleton ... ...

Abstract	Objective: To assess whether diet quality and specific dietary components are associated with hypertensive disorders of pregnancy (HDP). Study design: Nested case control study in a prospectively collected cohort of 450 participants with singleton pregnancies who completed the National Institutes of Health Diet Health Questionnaire II (DHQ-II) in the third trimester or within 3 months of delivery. Patients with fetal anomalies, conception by in-vitro fertilization, and deliveries at outside hospitals were excluded from the original prospective cohort study. Cases were patients diagnosed with HDP and controls were patients without HDP. Cases and controls were matched by BMI class in a 1:2 ratio. Exposures of interest were HEI-2015 score components and other DHQ-II dietary components including minerals, caffeine, and water. These dietary components were compared between cohorts using univariate analyses. Main outcome measures: HEI-2015 total scores representing diet quality, component scores, and objective background data between patients with HDP and patients without HDP. Results: 150 patients with HDP were matched to 300 controls without HDP. Baseline demographics were similar between groups, including BMI. Patients with HDP were less likely to have high quality diets (HEI ≥ 70) than controls (7.3 % v 15.7 %, P = 0.02). HDP were associated with significantly higher dairy, saturated fat, and sodium intake compared to controls. Other components were similar between groups. Conclusion: Patients with HDP are more likely to have lower diet quality and higher consumption of sodium, dairy, and saturated fats. These results can be used to study antenatal diet modification in patients at high risk of HDP.
Language	English
Publishing date	2024-03-18
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2584464-7
ISSN	2210-7797 ; 2210-7789
ISSN (online)	2210-7797
ISSN	2210-7789
DOI	10.1016/j.preghy.2024.101120
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