Placenta and Trophoblast Volume 2 Methods And Protocols (Methods in Molecular Medicine)

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In humans, taurine is maternally derived during pregnancy as fetal production is insufficient to support development [60]. Maternal taurine deprivation in animal models demonstrates decreased fertility and increased incidence of fetal resorption and stillbirth in feline and rodent models [59] , [65] — [67]. Surviving neonates have lower birth weight, decreased brain weight and experience slower growth [59] , [60]. In the complete absence of taurine transporters, knock-out mice are significantly growth restricted at birth, and have visual, auditory, and muscular alterations in adulthood compared to their wild types [68].

At 40 mM alcohol, the inhibition of taurine transporter function is potentially exposing the fetus to periods of significant deprivation [73]. Thus taurine deprivation, secondary to ethanol exposure, may contribute towards the varying degrees of neurological and behavioural differences and growth restriction seen in FASD affected children.

In addition, intracellular taurine behaves as an osmolyte, regulating the osmotic balance during cellular hydration, which plays an essential role in balancing proliferation and apoptosis [74] , [75]. Addition of taurine in vitro has also shown to increase rat neural stem cell proliferation and secretion of synapse developmental proteins [77]. When utilised as a dietary supplement after ethanol exposure in animal models, taurine markedly reduces the rate of abnormal neuronal migration and decreases the number of brain lesions [78].

Further work is required to investigate whether supplementation can improve outcomes where high levels of alcohol exposure are known to have occurred. Acetaldehyde, on the other hand, did not affect taurine transporter activity. Acetaldehyde has been suggested to enhance toxicity of ethanol and be the more toxic teratogen [27] , [30].

These pharmacological concentrations of acetaldehyde would not be found in a biological system as the metabolism of acetaldehyde plateaus at The system A amino acid transporter is a sodium-dependent transport system of small neutral amino acids that has shown to be associated with FGR, a key feature of FASD [6]. In contrast to the taurine transporter, no effects of ethanol on the activity of system A amino acid transport in either BeWo cells or in first trimester placental explants were observed, in agreement with previous work, with in vitro models, using placental explants exposed to 60 mM ethanol [26] , [80].

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Acetaldehyde, applied at concentrations much lower than in other studies [28] , [66] , [81] induced a small increase in 14 C-MeAIB uptake in placental explants, but not in BeWo cells. This observation may reflect a compensatory increase in response to an adverse environment as seen in other physiological systems when exposed to a chemical stimulus, as first trimester explants retain their phenotype in culture [82]. There is a potential capacity for functional compensation in first trimester tissue, but not in cells, because of the diversity of cell types. As a trophoblast cell layer model, BeWo cells are commonly used to assess membrane transport as they express similar functional transport receptors [83] — [87] and retain the trophoblastic property of fusion into a multinucleated layer [88].

Detrimental effects have been described at pharmacological concentrations of acetaldehyde x20 to greater than those found biologically on system A transport of the amino acid, valine, in rat placental explants [89] and on the neutral system A transport of lithium-dependant L-alanine in human term explants [79]. This indicates that acetaldehyde may have selective effects on different amino acid transporters; however, though toxic at pharmacological concentrations, it does not affect MeAIB transport at clinically relevant levels. Alcohol consumption is common in many cultures [90] and alcohol over-use by young women is an increasing public health issue [1].

This study has shown that short, acute exposure to ethanol, at levels readily achieved in a single sitting, negatively affects first trimester placental cell growth, trophoblast migration and the function of an amino acid transporter vital for normal neurological development. This implicates adverse developmental effects of ethanol at the earliest stages of placental development, with potentially major developmental effects for the fetus, and suggests that abstinence in the early stages of pregnancy is the safest clinical advice.

We would also like to express our gratitude to Dr A. Berry, Mrs C.

Meet the placenta! - Reproductive system physiology - NCLEX-RN - Khan Academy

Vinod and Mrs L. Abi for their assistance in obtaining first trimester placentas. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. National Center for Biotechnology Information , U. PLoS One. Published online Feb 4. Jones , 1 , 2 Nathalie J. Robinson , 1 , 2 Susan L. Greenwood , 1 , 2 John D. Aplin , 1 , 2 and Clare L.

Tower 1 , 2.

Phenotypic Analysis of the Rat Placenta

Rebecca L. Nathalie J. Susan L. John D.

Publications — Reproductive and Cardiovascular Disease Research Group

Clare L. Yan-Ling Wang, Editor. Author information Article notes Copyright and License information Disclaimer. Competing Interests: The authors have declared that no competing interests exist. Received Sep 18; Accepted Dec This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

This article has been cited by other articles in PMC. Abstract Fetal alcohol spectrum disorder FASD describes developmental issues from high maternal alcohol intake, which commonly results in fetal growth restriction and long term morbidity.

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  • Placenta and Trophoblast: Methods and Protocols, Volume II / Edition 1.

Introduction Rates of alcohol consumption amongst women of reproductive age are steadily increasing, with almost half of all young women in the UK are reported to drink during the week and a fifth reported to binge drink [1]. Immunohistochemistry Quantification Cell turnover was quantified by the ratio of positively stained cells and total number of nuclei for Ki67 in-cycle and M30 cytokeratin epitope to generate proliferative and apoptotic indices respectively. Open in a separate window. Figure 1. Effect of ethanol and acetaldehyde on invasive capacity of SGHPL4 extravillous cytotrophoblast cells.

Figure 2. Effect of ethanol and acetaldehyde on the proliferation and apoptotic index of BeWo cells. Figure 3. Effect of ethanol and acetaldehyde on the proliferation and apoptotic index of first trimester tissue. Figure 4. Figure 5.

Methods and Protocols, Volume I

Discussion The first trimester of pregnancy is crucial for placental development, which in turn provides for organogenesis and fetal growth [41]. References 1. NHS The health and social care information centre. Br J Addict 84 : — The generation R study.


Ann Epidemiol 17 : — Int J Epidemiol 39 : — BMC Public Health 11 : British Medical Association Fetal alcohol spectrum disorders - A guide for healthcare professionals. British Medical Association Board of Science. Maulik D Fetal growth restriction: the etiology. Clin Obstet Gynecol 49 : — Am J Obstet Gynecol : — Br J Obstet Gynaecol 93 : — Khong TY Placental vascular development and neonatal outcome.

Semin Neonatol 9 : — Pediatr Res 58 : — Placenta 29 : — Health Rep 6 : 45— Hypertension 36 : — Espinet C, Argiles JM Ethanol and acetaldehyde concentrations in the rat foeto-maternal system after an acute ethanol administration given to the mother. Arch Int Physiol Biochim 92 : — Guerri C, Sanchis R Acetaldehyde and alcohol levels in pregnant rats and their fetuses.

Alcohol 2 : — Burd L, Blair J, Dropps K Prenatal alcohol exposure, blood alcohol concentrations and alcohol elimination rates for the mother, fetus and newborn. J Perinatol 32 : — Alcoholism-Clinical and Experimental Research 10 : 39— Cell Biochem Funct 23 : — Beijing Da Xue Xue Bao 37 : — Alcohol 44 : — Reprod Fertil Dev 24 : — J Pediatr Gastroenterol Nutr 4 : — Rout UK, Dhossche JM Liquid-diet with alcohol alters maternal, fetal and placental weights and the expression of molecules involved in integrin signaling in the fetal cerebral cortex. Prog Biochem Pharmacol 18 : — Life Sci 29 : — Ther Drug Monit 23 : — Toxicol In Vitro 24 : 56— Teratology 36 : — Exp Pathol 33 : 17— PLoS One 6 : e Fisher SE, Atkinson M, Van Thiel DH Selective fetal malnutrition: the effect of nicotine, ethanol, and acetaldehyde upon in vitro uptake of alpha-aminoisobutyric acid by human term placental villous slices.

Dev Pharmacol Ther 7 : — Alcohol 9 : — Alcohol Clin Exp Res 20 : — J Soc Gynecol Investig 13 : — Sibley CP Understanding placental nutrient transfer—why bother? New biomarkers of fetal growth. J Physiol : — Sturman JA Taurine in development. Physiol Rev 73 : — J Cell Sci Pt 2 : — Placenta 32 : — Greenwood SL, Sibley CP In vitro methods for studying human placental amino acid transport: placental villous fragments.

Jones CJ, Aplin JD Glycosylation at the fetomaternal interface: does the glycocode play a critical role in implantation? Glycoconj J 26 : — Difference between alcoholic and nonalcoholic subjects. N Engl J Med : — Hayashi M Ethanol and acetaldehyde concentrations in pregnant rats after administration of ethanol. Arukoru Kenkyuto Yakubutsu Ison 26 : 89— Birth Defects Orig Artic Ser 18 : 89— Viability of the tissue and active CT doubling was demonstrated by the incorporation of BrDU in some cells over 24 hours of culture.

Magnified areas are indicated in the hatched boxes.


More unexpectedly, we observed a similar effect of ABCB1 knockdown on vasculogenesis and syncytialization of primary EVT cells in both explants and microdissected EVT anchoring columns. This process usually takes 72 hours in untreated explants which will shed and spontaneously generate a new syncytium in appropriate culture conditions. In contrast, the PTC placenta, like the term placenta in Fig. Endoreduplication events cannot be excluded as the nuclei of these cells are also enlarged. Excessive decidual inflammation during the early events of EVT invasion is a contributory factor in shallow placentation, abruption and PE.

The authors wish to thank Dr.

The authors have declared that no conflict of interest exists. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries other than missing content should be directed to the corresponding author for the article. Volume 22 , Issue The full text of this article hosted at iucr. If you do not receive an email within 10 minutes, your email address may not be registered, and you may need to create a new Wiley Online Library account.

If the address matches an existing account you will receive an email with instructions to retrieve your username. Caroline E. Dunk Corresponding Author E-mail address: dunk lunenfeld. Search for more papers by this author. Jane J. Stephen J. Stephen G. Dunk and Pappas contributed equally to this manuscript.

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Share Give access Share full text access. Share full text access. Please review our Terms and Conditions of Use and check box below to share full-text version of article. Gestational age at delivery weeks Mean Range All clinical data were collected from the patient records by the Placental Biobank of Mount Sinai Hospital, Toronto, with ethical approval from the Mount Sinai Research Ethics Board and written patient consent. All primers were blasted using NCBI primer blast to ensure specificity to the gene of interest and confirmed to produce a single band of the expected size by PCR analysis.

All antibodies used in the study were optimized for antigen retrieval and dilution. Figure 1 Open in figure viewer PowerPoint. B and D, Sense controls in serial sections showing no staining. H, Mouse IgG control. Figure 2 Open in figure viewer PowerPoint. Data were normalized as in A. Figure 3 Open in figure viewer PowerPoint. Data were normalized as in Figure 2 A. Graphs show the mean and standard deviation. Figure 4 Open in figure viewer PowerPoint. Formation of a new ST layer on the villous surface can also be observed in the CK8 image red arrows. Magnified images shown in Bi and Bii correspond to the hatched boxes.

Negative isotype controls of guinea pig and rabbit IgG or rabbit and mouse IgG showed no staining. Three placentas were used in experimental triplicates. Figure 5 Open in figure viewer PowerPoint. Google Scholar.


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