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      Mapping the expression of transient receptor potential channels across murine placental development

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          Abstract

          Transient receptor potential (TRP) channels play prominent roles in ion homeostasis by their ability to control cation influx. Mouse placentation is governed by the processes of trophoblast proliferation, invasion, differentiation, and fusion, all of which require calcium signaling. Although certain TRP channels have been shown to contribute to maternal–fetal transport of magnesium and calcium, a role for TRP channels in specific trophoblast functions has been disregarded. Using qRT-PCR and in situ hybridisation, the spatio-temporal expression pattern of TRP channels in the mouse placenta across gestation (E10.5–E18.5) was assessed. Prominent expression was observed for Trpv2, Trpm6, and Trpm7. Calcium microfluorimetry in primary trophoblast cells isolated at E14.5 of gestation further revealed the functional activity of TRPV2 and TRPM7. Finally, comparing TRP channels expression in mouse trophoblast stem cells (mTSCs) and mouse embryonic stem cells (mESC) confirmed the specific expression of TRPV2 during placental development. Moreover, TRP channel expression was similar in mTSCs compared to primary trophoblasts and validate mTSC as a model to study TRP channels in placental development. Collectivity, our results identify a specific spatio-temporal TRP channel expression pattern in trophoblasts, suggesting a possible involvement in regulating the process of placentation.

          Supplementary Information

          The online version contains supplementary material available at 10.1007/s00018-021-03837-3.

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          RNAscope: a novel in situ RNA analysis platform for formalin-fixed, paraffin-embedded tissues.

          In situ analysis of biomarkers is highly desirable in molecular pathology because it allows the examination of biomarker status within the histopathological context of clinical specimens. Immunohistochemistry and DNA in situ hybridization (ISH) are widely used in clinical settings to assess protein and DNA biomarkers, respectively, but clinical use of in situ RNA analysis is rare. This disparity is especially notable when considering the abundance of RNA biomarkers discovered through whole-genome expression profiling. This is largely due to the high degree of technical complexity and insufficient sensitivity and specificity of current RNA ISH techniques. Here, we describe RNAscope, a novel RNA ISH technology with a unique probe design strategy that allows simultaneous signal amplification and background suppression to achieve single-molecule visualization while preserving tissue morphology. RNAscope is compatible with routine formalin-fixed, paraffin-embedded tissue specimens and can use either conventional chromogenic dyes for bright-field microscopy or fluorescent dyes for multiplex analysis. Unlike grind-and-bind RNA analysis methods such as real-time RT-PCR, RNAscope brings the benefits of in situ analysis to RNA biomarkers and may enable rapid development of RNA ISH-based molecular diagnostic assays. Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
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            TRP channels as cellular sensors.

            TRP channels are the vanguard of our sensory systems, responding to temperature, touch, pain, osmolarity, pheromones, taste and other stimuli. But their role is much broader than classical sensory transduction. They are an ancient sensory apparatus for the cell, not just the multicellular organism, and they have been adapted to respond to all manner of stimuli, from both within and outside the cell.
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              Growth and function of the normal human placenta.

              The placenta is the highly specialised organ of pregnancy that supports the normal growth and development of the fetus. Growth and function of the placenta are precisely regulated and coordinated to ensure the exchange of nutrients and waste products between the maternal and fetal circulatory systems operates at maximal efficiency. The main functional units of the placenta are the chorionic villi within which fetal blood is separated by only three or four cell layers (placental membrane) from maternal blood in the surrounding intervillous space. After implantation, trophoblast cells proliferate and differentiate along two pathways described as villous and extravillous. Non-migratory, villous cytotrophoblast cells fuse to form the multinucleated syncytiotrophoblast, which forms the outer epithelial layer of the chorionic villi. It is at the terminal branches of the chorionic villi that the majority of fetal/maternal exchange occurs. Extravillous trophoblast cells migrate into the decidua and remodel uterine arteries. This facilitates blood flow to the placenta via dilated, compliant vessels, unresponsive to maternal vasomotor control. The placenta acts to provide oxygen and nutrients to the fetus, whilst removing carbon dioxide and other waste products. It metabolises a number of substances and can release metabolic products into maternal and/or fetal circulations. The placenta can help to protect the fetus against certain xenobiotic molecules, infections and maternal diseases. In addition, it releases hormones into both the maternal and fetal circulations to affect pregnancy, metabolism, fetal growth, parturition and other functions. Many placental functional changes occur that accommodate the increasing metabolic demands of the developing fetus throughout gestation.
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                Author and article information

                Contributors
                joris.vriens@kuleuven.be
                Journal
                Cell Mol Life Sci
                Cell Mol Life Sci
                Cellular and Molecular Life Sciences
                Springer International Publishing (Cham )
                1420-682X
                1420-9071
                21 April 2021
                21 April 2021
                2021
                : 78
                : 11
                : 4993-5014
                Affiliations
                [1 ]GRID grid.5596.f, ISNI 0000 0001 0668 7884, Laboratory of Endometrium, Endometriosis and Reproductive Medicine, Department of Development and Regeneration, , KU Leuven, ; Herestraat 49, Box 611, 3000 Leuven, Belgium
                [2 ]GRID grid.5596.f, ISNI 0000 0001 0668 7884, Laboratory of Ion Channel Research, Department of Cellular and Molecular Medicine, VIB Center for Brain and Disease Research, , KU Leuven, ; Herestraat 49, Box 802, 3000 Leuven, Belgium
                [3 ]GRID grid.5335.0, ISNI 0000000121885934, Department of Physiology, Development and Neuroscience, Centre for Trophoblast Research, , University of Cambridge, ; Downing Street, Cambridge, CB2 3EG UK
                [4 ]GRID grid.418195.0, ISNI 0000 0001 0694 2777, Epigenetics Programme, , The Babraham Institute, Babraham Research Campus, ; Cambridge, CB22 3AT UK
                [5 ]GRID grid.418274.c, ISNI 0000 0004 0399 600X, Centro de Investigación Principe Felipe, , Molecular Basis of Human Diseases, ; Eduardo Primo Yúfera 3, 46012 Valencia, Spain
                [6 ]GRID grid.16563.37, ISNI 0000000121663741, Dipartimento di Scienze del Farmaco, , Universita degli Studi del Piemonte Orientale Amedeo, ; Vercelli, 13100 Italy
                Author information
                http://orcid.org/0000-0002-2502-0409
                Article
                3837
                10.1007/s00018-021-03837-3
                8233283
                33884443
                79d1a830-81e5-4b7f-b0b3-0e7a0fcc9f46
                © The Author(s) 2021

                Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 2 December 2020
                : 17 March 2021
                : 8 April 2021
                Funding
                Funded by: Research foundations Flanders
                Award ID: 12R4519N
                Award Recipient :
                Funded by: Research Foundation Flanders
                Award ID: G.0D1417N, G.084515N, G.0A6719N
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100004497, Onderzoeksraad, KU Leuven;
                Funded by: Centre of Trophoblast Research, University of Cambridge
                Categories
                Original Article
                Custom metadata
                © Springer Nature Switzerland AG 2021

                Molecular biology
                trp channels,placental development,primary trophoblast cells,trophoblast stem cells

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