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Expression of Selected Connexin and Aquaporin Genes and Real-Time Proliferation of Porcine Endometrial Luminal Epithelial Cells in Primary Culture Model

K. Wojtanowicz-Markiewicz, M. Kulus, S. Knap, I. Kocherova, M. Jankowski, K. Stefańska, M. Jeseta, H. Piotrowska-Kempisty, D. Bukowska, M. Zabel, P. Mozdziak, M. Nowicki, B. Kempisty, P. Antosik,

. 2020 ; 2020 (-) : 7120375. [pub] 20200203

Language English Country United States

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc20028525

Luminal epithelial cells are the first embryonic-maternal contact site undergoing very specific changes associated with reproductive processes. Cells prepare for embryo development by increasing their volume, with the help of aquaporins that provide a transcellular path of rapid water movement during the secretion and absorption of fluids, as well as connexins enabling the flow of inorganic ions and small molecules. In this work, we have examined how AQPs and Cx's behave in luminal epithelium primary cell culture. Cells obtained from porcine specimen during slaughter were primarily in vitro cultured for 7 days. Their proliferation patterns were then analyzed using RTCA, with the expression of genes of interest evaluated with the use of immunofluorescence and RT-qPCR. The results of these changes of gene of interest expression were analyzed on each of the seven days of the porcine luminal primary cell culture. Our study showed that the significant changes were noted in the case of Cx43, whose level of protein expression and distribution increases after 120 hours of culture, when the cells enter the lag phase, and maintains an upward trend until the end of the culture. We noted an increase in AQP4, AQP7, AQP8, and AQP11 levels throughout the entire culture period, while the largest differences in expression were found in AQP3, AQP4, and AQP10. The obtained results could become a point of reference for further in vivo and clinical research. Experiments conducted with these proteins showed that they influence the endometrial fluid content during the oestrous cycle and participate in the process of angiogenesis, which intensifies during endometrial development.

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$a Luminal epithelial cells are the first embryonic-maternal contact site undergoing very specific changes associated with reproductive processes. Cells prepare for embryo development by increasing their volume, with the help of aquaporins that provide a transcellular path of rapid water movement during the secretion and absorption of fluids, as well as connexins enabling the flow of inorganic ions and small molecules. In this work, we have examined how AQPs and Cx's behave in luminal epithelium primary cell culture. Cells obtained from porcine specimen during slaughter were primarily in vitro cultured for 7 days. Their proliferation patterns were then analyzed using RTCA, with the expression of genes of interest evaluated with the use of immunofluorescence and RT-qPCR. The results of these changes of gene of interest expression were analyzed on each of the seven days of the porcine luminal primary cell culture. Our study showed that the significant changes were noted in the case of Cx43, whose level of protein expression and distribution increases after 120 hours of culture, when the cells enter the lag phase, and maintains an upward trend until the end of the culture. We noted an increase in AQP4, AQP7, AQP8, and AQP11 levels throughout the entire culture period, while the largest differences in expression were found in AQP3, AQP4, and AQP10. The obtained results could become a point of reference for further in vivo and clinical research. Experiments conducted with these proteins showed that they influence the endometrial fluid content during the oestrous cycle and participate in the process of angiogenesis, which intensifies during endometrial development.
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$a Kulus, Magdalena $u Veterinary Centre, Nicolaus Copernicus University in Torun, Torun, Poland.
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$a Knap, Sandra $u Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland.
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$a Kocherova, Ievgenia $u Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland.
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$a Jankowski, Maurycy $u Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland.
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$a Stefańska, Katarzyna $u Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland.
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$a Jeseta, Michal $u Department of Obstetrics and Gynaecology, University Hospital and Masaryk University, Brno, Czech Republic.
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$a Zabel, Maciej $u Division of Histology and Embryology, Department of Human Morphology and Embryology, Wroclaw Medical University, Wroclaw, Poland. Division of Anatomy and Histology, University of Zielona Góra, Zielona Góra, Poland.
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$a Mozdziak, Paul $u Physiology Graduate Program, North Carolina State University, Raleigh, North Carolina, USA.
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$a Nowicki, Michał $u Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland.
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$a Kempisty, Bartosz $u Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland. Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland. Department of Obstetrics and Gynaecology, University Hospital and Masaryk University, Brno, Czech Republic.
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