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      Human Cumulus Cells in Long-Term In Vitro Culture Reflect Differential Expression Profile of Genes Responsible for Planned Cell Death and Aging—A Study of New Molecular Markers

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          Abstract

          In the ovarian follicle, maturation of the oocyte increases in the presence of somatic cells called cumulus cells (CCs). These cells form a direct barrier between the oocyte and external environment. Thanks to bidirectional communication, they have a direct impact on the oocyte, its quality and development potential. Understanding the genetic profile of CCs appears to be important in elucidating the physiology of oocytes. Long-term in vitro culture of CCs collected from patients undergoing controlled ovarian stimulation during in vitro fertilization procedure was conducted. Using microarray expression analysis, transcript levels were assessed on day 1, 7, 15, and 30 of culture. Apoptosis and aging of CCs strictly influence oocyte quality and subsequently the outcome of assisted reproductive technologies (ART). Thus, particular attention was paid to the analysis of genes involved in programmed cell death, aging, and apoptosis. Due to the detailed level of expression analysis of each of the 133 analyzed genes, three groups were selected: first with significantly decreased expression during the culture; second with the statistically lowest increase in expression; and third with the highest significant increase in expression. COL3A1, SFRP4, CTGF, HTR2B, VCAM1, TNFRSF11B genes, belonging to the third group, were identified as potential carriers of information on oocyte quality.

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          Most cited references59

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          Oocyte environment: follicular fluid and cumulus cells are critical for oocyte health.

          Bidirectional somatic cell-oocyte signaling is essential to create a changing intrafollicular microenvironment that controls primordial follicle growth into a cohort of growing follicles, from which one antral follicle is selected to ovulate a healthy oocyte. Such intercellular communications allow the oocyte to determine its own fate by influencing the intrafollicular microenvironment, which in turn provides the necessary cellular functions for oocyte developmental competence, which is defined as the ability of the oocyte to complete meiosis and undergo fertilization, embryogenesis, and term development. These coordinated somatic cell-oocyte interactions attempt to balance cellular metabolism with energy requirements during folliculogenesis, including changing energy utilization during meiotic resumption. If these cellular mechanisms are perturbed by metabolic disease and/or maternal aging, molecular damage of the oocyte can alter macromolecules, induce mitochondrial mutations, and reduce adenosine triphosphate production, all of which can harm the oocyte. Recent technologies are now exploring transcriptional, translational, and post-translational events within the human follicle with the goal of identifying biomarkers that reliably predict oocyte quality in the clinical setting.
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            Mater, a maternal effect gene required for early embryonic development in mice.

            Maternal effect genes produce mRNA or proteins that accumulate in the egg during oogenesis. We show here that Mater, a mouse oocyte protein dependent on the maternal genome, is essential for embryonic development beyond the two-cell stage. Females lacking the maternal effect gene Mater are sterile. Null males are fertile.
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              The gametic synapse: RNA transfer to the bovine oocyte.

              Even after several decades of quiescent storage in the ovary, the female germ cell is capable of reinitiating transcription to build the reserves that are essential to support early embryonic development. In the current model of mammalian oogenesis, there exists bilateral communication between the gamete and the surrounding cells that is limited to paracrine signaling and direct transfer of small molecules via gap junctions existing at the end of the somatic cells' projections that are in contact with the oolemma. The purpose of this work was to explore the role of cumulus cell projections as a means of conductance of large molecules, including RNA, to the mammalian oocyte. By studying nascent RNA with confocal and transmission electron microscopy in combination with transcript detection, we show that the somatic cells surrounding the fully grown bovine oocyte contribute to the maternal reserves by actively transferring large cargo, including mRNA and long noncoding RNA. This occurrence was further demonstrated by the reconstruction of cumulus-oocyte complexes with transfected cumulus cells transferring a synthetic transcript. We propose selective transfer of transcripts occurs, the delivery of which is supported by a remarkable synapselike vesicular trafficking connection between the cumulus cells and the gamete. This unexpected exogenous contribution to the maternal stores offers a new perspective on the determinants of female fertility. © 2014 by the Society for the Study of Reproduction, Inc.
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                Author and article information

                Journal
                Cells
                Cells
                cells
                Cells
                MDPI
                2073-4409
                21 May 2020
                May 2020
                : 9
                : 5
                : 1265
                Affiliations
                [1 ]Division of Infertility and Reproductive Endocrinology, Department of Gynecology, Obstetrics and Gynecological Oncology, Poznan University of Medical Sciences, 33 Polna St., 60-535 Poznan, Poland; blazej.chermula@ 123456wp.pl (B.C.); pawelczyk.leszek@ 123456ump.edu.pl (L.P.); rspaczynski@ 123456yahoo.com (R.Z.S.)
                [2 ]Department of Anatomy, Poznan University of Medical Sciences, 6 Swiecickiego St., 60-781 Poznan, Poland; wkranc@ 123456ump.edu.pl (W.K.); g.hutchings.16@ 123456abnd.ac.uk (G.H.); krzysztof.janowicz.16@ 123456abdn.ac.uk (K.J.)
                [3 ]Department of Histology and Embryology, Poznan University of Medical Sciences, 6 Swiecickiego St., 60-781 Poznan, Poland; karoljopek01@ 123456gmail.com (K.J.); joanna.budna@ 123456wp.pl (J.B.-T.); claudia.dompe.16@ 123456abdn.ac.uk (C.D.); l.moncrieff.16@ 123456abdn.ac.uk (L.M.)
                [4 ]The School of Medicine, Medical Sciences and Nutrition, University of Aberdeen, Aberdeen AB25 2ZD, UK
                [5 ]Department of Toxicology, Poznan University of Medical Sciences, 30 Dojazd St., 60-631 Poznan, Poland; malgorzata.jozkowiak@ 123456gmail.com
                [6 ]Department of Obstetrics and Gynecology, University Hospital and Masaryk University, 20 Jihlavská St., 62500 Brno, Czech Republic; jeseta@ 123456gmail.com
                [7 ]Prestage Department of Poultry Science, North Carolina State University, Raleigh, NC 27695, USA; jnppo@ 123456ncsu.edu
                [8 ]Physiology Graduate Program, North Carolina State University, Raleigh, NC 27695, USA; pemozdzi@ 123456ncsu.edu
                [9 ]Department of Veterinary Surgery, Institute of Veterinary Medicine, Nicolaus Copernicus University in Torun, 1 Lwowska St., 87-100 Torun, Poland
                Author notes
                [* ]Correspondence: bkempisty@ 123456ump.edu.pl ; Tel.: +48-6185-464-18
                Author information
                https://orcid.org/0000-0001-7518-4834
                https://orcid.org/0000-0001-9285-5417
                https://orcid.org/0000-0003-1700-1627
                https://orcid.org/0000-0003-1778-3454
                https://orcid.org/0000-0003-2764-6007
                https://orcid.org/0000-0002-1575-3123
                https://orcid.org/0000-0002-4306-6590
                Article
                cells-09-01265
                10.3390/cells9051265
                7291080
                32455542
                9de7355a-65e8-429f-9f6e-d4251df57df6
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 25 March 2020
                : 19 May 2020
                Categories
                Article

                cumulus cells,human,programmed cell death,gene expression

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