Gestational Age-Dependent Fetal Fluid Dynamics in the Ovine Developmental Model: Establishment of Surrogate Markers for the Differentiation of Stem Cell Origin

The ovine developmental model represents the standard in vivo model for studies involving maternofetal physiology, amniotic fluid (AF) research, and fetal cell therapy prior to human clinical use. Although being close to the human fetal anatomy, 2 separate extraembryonic fluid compartments remain during gestation, known as the amnion and the allantois. A clear distinction between AF versus allantoic fluid (AL) is therefore indispensable for correct scientific conclusions with regard to human translation. In the presented study, the biochemical composition of AF and AL was evaluated in ovine gravid uteri postmortem (n = 31) over the entire gestation. Four parameters, consisting of Na+, Cl–, Mg2+, and total protein, have been found to allow for specific discrimination of the 2 fetal fluids at all gestational phases and therefore as potential surrogate parameters for gestational age. In addition, volumetric changes of the developing fetus and the 2 fetal fluid cavities were analyzed by contrast-enhanced computed tomography (n = 12). AF showed a significant, linear volumetric increase over gestation, whereas AL volume maintained relatively static independent of gestational age. These results serve as a basis for future studies by providing surrogate markers enabling a reliable distinction of isolated fetal fluids and contained cells in the ovine developmental model over the entire gestation.

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Fetal-fluid proteome analyses in late-term healthy pregnant mares and in mares with experimentally induced ascending placentitis

Reproduction Fertility and Development ◽

10.1071/rd18460 ◽

2019 ◽

Vol 31 (9) ◽

pp. 1486 ◽

Cited By ~ 2

Author(s):

Igor F. Canisso ◽

Shavahn Loux ◽

Kirsten E. Scoggin ◽

Edward L. Squires ◽

Mats H. Troedsson ◽

...

Keyword(s):

Calcium Binding ◽

Protein Composition ◽

Allantoic Fluid ◽

Signalling Molecules ◽

Disease States ◽

Streptococcus Equi ◽

Before And After ◽

Fetal Fluid ◽

Fetal Fluids ◽

Experimentally Induced

Characterisation of fetal fluids in healthy and disease states of pregnant mares can help to unravel the pathophysiology and to identify putative markers of disease. Thus, this study aimed to compare the protein composition of: (1) amniotic and allantoic fluids of healthy mares obtained immediately after euthanasia and (2) allantoic fluid harvested via centesis before and after experimental induction of placentitis via transcervical inoculation of Streptococcus equi ssp zooepidemicus in healthy mares. Fetal fluids were analysed with a high-throughput proteomic technique after in-gel digestion. Statistical comparisons were performed following normalisation of peptide spectral match. Global normalisation was performed to calculate relative expression. There were 112 unique proteins present in both allantoic and amniotic fluids. There were 13 and 29 proteins defined as amniotic- or allantoic-specific respectively that were present in at least two fluid samples. Another 26 proteins were present in both amniotic and allantoic fluids. Panther DB functional classification grouped fetal-fluid proteins as transfer carriers, signalling molecules, receptors, immunity, hydrolase, enzymes, membrane traffic, cytoskeleton, cell adhesion, calcium binding and extracellular matrix. Experimentally induced placentitis resulted in 10 proteins being upregulated and 10 downregulated in allantoic fluid. Newly identified proteins and changes in the fetal-fluid proteome provide clues about the physiology of pregnancy and pathogenesis of placentitis.

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54 COMPOSITION OF ALLANTOIC FLUID IN CATTLE PREGNANT WITH AI-, IVP-, OR NUCLEAR TRANSFER-GENERATED EMBRYOS

Reproduction Fertility and Development ◽

10.1071/rdv17n2ab54 ◽

2005 ◽

Vol 17 (2) ◽

pp. 177 ◽

Cited By ~ 2

Author(s):

C. Morrow ◽

M. Berg ◽

R. McDonald ◽

D. Wells ◽

A. Peterson ◽

...

Keyword(s):

Nuclear Transfer ◽

Allantoic Fluid ◽

Diagnostic Markers ◽

Morphological Data ◽

Fluid Composition ◽

Abnormal Placentation ◽

Fetal Fluid ◽

Estrone Sulphate ◽

Early Diagnostic ◽

Fetal Fluids

Abnormal placentation, pregnancy failure, and hydroallantois are associated with somatic cell nuclear transfer (SCNT) in cattle. Identification of diagnostic markers for abnormal placentation in early gestation would permit therapeutic intervention. Ultrasonography and transvaginal sampling of amniotic and/or allantoic (fetal) fluid enables regular monitoring of fetal health. We report on the composition of serial samples of fetal fluid from individual cows between Days 70–130 of gestation and the potential of steroid and electrolyte composition as an early diagnostic marker for the subsequent occurrence of hydroallantois in SCNT pregnancies in cattle. On Day 70, pregnancy rates were 50% and 60% for cows or heifers implanted with single in vitro-fertilized (IVP, 20/40) or SCNT (25/42) embryos, respectively, and 67% for pregnancies generated by artificial insemination (AI, 12/18). Resulting fetuses were either clones (SCNT) or offspring (IVP/AI) of a donor Holstein bull. Fetal fluids, sampled using ultrasound-guided transvaginal puncture, were collected on Days 70, 100, and 130 of gestation (n = 12 and 139 for amniotic and allantoic samples, respectively). Placental and fetal morphological data were collected following slaughter between Days 135–163 of gestation (n = 14, 20, and 10 for SCNT, IVP, and AI groups, respectively). Fetal fluids were analyzed for progesterone, estrone sulphate, sodium, chloride, potassium, creatinine, urea, calcium, magnesium and phosphate. Pregnancy outcomes for the SCNT group were retrospectively classified as: Fail 100 (pregnancies failing between Days 70–99; n = 6); Fail 130 (failing between Days 100–129; n = 5); Hydrops (greater than 10 L combined amniotic and allantoic fluid at postmortem between Days 135–163; n = 8) and SCNT Pregnant 150 (pregnant between Days 135–163; n = 6). IVP and AI pregnancies were classified as IVP or AI Pregnant 150. Fluid composition was analyzed by ANOVA on log-transformed data. On Day 70, allantoic progesterone and estrone sulphate concentrations were significantly higher (P < 0.05) for the SCNT cows compared to the IVP/AI Pregnant 150 cows. On Day 70, allantoic potassium, chloride, creatinine, and urea concentrations were significantly higher (P < 0.05) for the SCNT Hydrops cows compared to the IVP/AI Pregnant 150 cows. In addition, Day 70 allantoic creatinine and urea concentrations were significantly higher (P < 0.05) for the SCNT Hydrops cows compared to other SCNT groups. By Day 100, allantoic chloride, creatinine, and urea concentrations in SCNT Hydrops cows were significantly lower (P < 0.05) than in IVP/AI Pregnant 150 groups. We conclude that elevated Day 70 allantoic urea and creatinine concentrations are potential early diagnostic markers predicting hydroallantois in recipient cattle carrying SCNT fetuses. Further investigation of these markers in other somatic donor cell lines used for nuclear transfer is warranted to determine their general utility.

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Gestational profile of the stimulatory effects of porcine amniotic and allantoic fluids on prostaglandin G/H synthase activity

Reproduction Fertility and Development ◽

10.1071/rd9900581 ◽

1990 ◽

Vol 2 (5) ◽

pp. 581

Author(s):

GE Rice ◽

MH Wong ◽

P Christensen ◽

V Dantzer ◽

E Skadhauge

Keyword(s):

Amniotic Fluid ◽

Maximum Dose ◽

Allantoic Fluid ◽

Late Gestation ◽

Dependent Manner ◽

Stimulatory Activity ◽

Dose Dependent ◽

Fetal Fluids

The effects of porcine fetal fluids (amniotic and allantoic) on microsomal prostaglandin G/H synthase (PGHS) activity were assessed. Both amniotic and allantoic fluids obtained from late-gestation sows stimulated PGHS activity (as indicated by increased formation of radiolabelled prostaglandin) in a dose-dependent manner. At the maximum dose tested, amniotic and allantoic fluids stimulated prostaglandin (PG) formation by 55.5 +/- 1.5 and 58.5 +/- 4.7%, respectively (n = 3, P less than 0.01). Based upon ED50 values, amniotic fluid was approximately threefold more effective than allantoic fluid in stimulating PG formation. The stimulatory effect of amniotic but not allantoic fluid increased significantly (P less than 0.01) during gestation (Days 47-112). The observed changes in the stimulatory effect of amniotic fluid on microsomal PG formation parallels the in vivo changes that occur in intra-uterine PG synthesis. Amniotic fluid stimulatory activity may contribute to this gestational increase in PG synthesis.

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DEVELOPMENTAL CHANGES OF FREE AMINO ACIDS IN BOVINE FETAL FLUIDS WITH GESTATIONAL AGE AND THE INTERRELATIONSHIPS BETWEEN THE AMINO ACID CONCENTRATIONS IN THE FLUID COMPARTMENTS

Reproduction ◽

10.1530/jrf.0.0440437 ◽

1975 ◽

Vol 44 (3) ◽

pp. 437-444 ◽

Cited By ~ 11

Author(s):

A. L. BAETZ ◽

W. T. HUBBERT ◽

C. K. GRAHAM

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Gestational Age ◽

Free Amino Acids ◽

Free Amino ◽

Developmental Changes ◽

Fluid Compartments ◽

Amino Acid Concentrations ◽

Fetal Fluids

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Panel Discussion II: Usefulness of in vitro and in vivo Model Systems for Predicting the Adverse Public Health Outcome for Antimicrobial Drug Residues in Food

Microbial Ecology in Health and Disease ◽

10.1080/08910600050216174 ◽

2000 ◽

Vol 12 (3) ◽

pp. 45-53

Author(s):

Andrew Beaulieu ◽

Jacques Boisseau ◽

Carl Cerniglia ◽

Denis Corpet ◽

A. Haydée Fernández ◽

...

Keyword(s):

Public Health ◽

Health Outcome ◽

Panel Discussion ◽

Antimicrobial Drug ◽

Model Systems ◽

In Vivo Model ◽

Drug Residues ◽

Public Health Outcome

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P1735 Arginine supplementation and oxidative processes: in vivo model

European Heart Journal ◽

10.1016/s0195-668x(03)94873-1 ◽

2003 ◽

Vol 24 (5) ◽

pp. 330

Author(s):

A KOPFF

Keyword(s):

In Vivo Model ◽

Oxidative Processes ◽

Arginine Supplementation

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Establishment of an in vivo model for KCNJ5 dependent hyperaldosteronism

Experimental and Clinical Endocrinology & Diabetes ◽

10.1055/s-0035-1547718 ◽

2015 ◽

Vol 122 (03) ◽

Cited By ~ 2

Author(s):

U Lichtenauer ◽

PL Schmid ◽

A Oßwald ◽

I Renner-Müller ◽

M Reincke ◽

...

Keyword(s):

In Vivo Model

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An in vivo Model for the Assessment of Acute Fibrinolytic Capacity of the Endothelium

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657722 ◽

1997 ◽

Vol 78 (04) ◽

pp. 1242-1248 ◽

Cited By ~ 68

Author(s):

David E Newby ◽

Robert A Wright ◽

Christopher A Ludlam ◽

Keith A A Fox ◽

Nicholas A Boon ◽

...

Keyword(s):

Blood Flow ◽

Substance P ◽

Sodium Nitroprusside ◽

Plasma Concentrations ◽

In Vivo Model ◽

Forearm Circulation ◽

Von Willebrand ◽

Local Release ◽

Willebrand Factor

SummaryThe effects on blood flow and plasma fibrinolytic and coagulation parameters of intraarterial substance P, an endothelium dependent vasodilator, and sodium nitroprusside, a control endothelium independent vasodilator, were studied in the human forearm circulation. At subsystemic locally active doses, both substance P (2-8 pmol/min) and sodium nitroprusside (2-8 μg/min) caused dose-dependent vasodilatation (p <0.001 for both) without affecting plasma concentrations of PAI-1, von Willebrand factor antigen or factor VIII:C activity. Substance P caused local increases in t-PA antigen and activity (p <0.001) in the infused arm while sodium nitroprusside did not. At higher doses, substance P increased blood flow and t-PA concentrations in the noninfused arm. We conclude that brief, locally active and subsystemic infusions of intraarterial substance P cause a rapid and substantial local release of t-PA which appear to act via a flow and nitric oxide independent mechanism. This model should provide a useful and selective method of assessing the in vivo capacity of the forearm endothelium to release t-PA acutely.

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