Katarzyna Stefańska, Ievgenia Kocherova, Sandra Knap, Magdalena Kulus, Piotr Celichowski and Michal Jeseta
The oviduct is a part of female reproductive tract that is essential for successful fertilization and early embryo development. It is lined with epithelium consisting of two types of cells: ciliated and secretory. The primary function of ciliated oviductal epithelial cells (OECs) is to support the transport of gametes and embryos through the ovary, whereas secretory OECs produce components of the oviductal fluid. Undoubtedly, the oviductal epithelium plays a major part in the early aspects of pregnancy development, from providing an optimal environment for gametes and embryos to supporting fertilization. Therefore, our aim was to gain a better insight into the genetic changes underlying function of these cells. We have harvested OECs from crossbred gilts (n=45), at the age of about nine months and which displayed two regular estrous cycles, and established long-term primary culture of porcine OECs. Microarray analysis was utilized to determine differentially expressed genes during day 1, 7, 15 and 30 of cultivation, with our results revealing54 differentially expressed genes belonging to three ontology groups: „maintenance of location”, „maintenance of protein location” and „maintenance of protein location in cell”. Since the biochemistry and morphology of epithelial cells may change during long term cultivation, we conclude that our results are a reflection of these changes and help to shed a light on porcine OECs properties in in vitro environment.
Running title: Maintenance of cellular protein location in porcine epithelial oviductal cells
Sylwia Borys-Wójcik, Ievgenia Kocherova, Piotr Celichowski, Małgorzata Popis, Michal Jeseta, Dorota Bukowska, Paweł Antosik, Michał Nowicki and Bartosz Kempisty
A wide variety of mechanisms controlling oligomerization are observed. The dynamic nature of protein oligomerization is important for bioactivity control. The oocyte must undergo a series of changes to become a mature form before it can fully participate in the processes associated with its function as a female gamete. The growth of oocytes in the follicular environment is accompanied by surrounding somatic cumulus (CCs) and granulosa cells (GCs). It has been shown that oocytes tested before and after in vitro maturation (IVM) differ significantly in the transcriptomic and proteomic profiles. The aim of this study was to determine new proteomic markers for the oligomerization of porcine oocyte proteins that are associated with cell maturation competence. The Affymetrix microarray assay was performed to examine the gene expression profile associated with protein oligomerization in oocytes before and after IVM. In total, 12258 different transcriptomes were analyzed, of which 419 genes with lower expression in oocytes after IVM. We found 9 genes: GJA1, VCP, JUP, MIF, MAP3K1, INSR, ANGPTL4, EIF2AK3, DECR1, which were significantly down-regulated in oocytes after IVM (in vitro group) compared to oocytes analyzed before IVM (in vivo group). The higher expression of genes involved in the oligomerization of the protein before IVM indicates that they can be recognized as important markers of biological activation of proteins necessary for the further growth and development of pig embryos.
Katarzyna Stefańska, Sandra Knap, Magdalena Kulus, Ievgenia Kocherova, Piotr Celichowski, Michal Jeseta, Marie Machatkova, Dorota Bukowska and Paweł Antosik
Oxygen metabolism is crucial in establishing successful pregnancy, since excessive amount of reactive oxygen species (ROS) may exert deleterious effects on the developing embryo. There are several defense mechanisms against oxidative stress in the female reproductive tract, including production of antioxidant enzymes by oviductal epithelial cells (OECs). Undoubtedly, OECs play major part in female fertility and may also serve as an in vitro model of the oviduct. Therefore, the aim of this study was to investigate the expression of genes involved in oxygen metabolism. We have isolated OECs from oviducts of crossbred gilts (n=45) and maintained their in vitro culture for 30 days, collecting their RNA at days 1, 7, 15 and 30. The gene expression was determined with the use of Affymetrix® Porcine Gene 1.1 ST Array Strip. Our results revealed 166 differentially expressed genes belonging to four ontology groups: „cellular response to oxidative stress”, “cellular response to oxygen-containing compound”, “cellular response to oxygen levels” and “cellular response to reactive oxygen species”, most of which are also involved in other major processes in the organism. However, our findings provide a valuable insight into porcine reproductive biology and may be utilized in optimization of assisted reproduction techniques.
Running title: Genes involved in oxygen metabolism in oviductal epithelial cells
Edyta Porowska, Magdalena Kulus, Maurycy Jankowski, Ievgenia Kocherova, Michal Jeseta, Agata Chamier-Gliszczyńska, Katarzyna Stefańska, Blanka Borowiec, Dorota Bukowska, Klaus P. Brüssow, Bartosz Kempisty and Paweł Antosik
Pyometra is the most common gynecological disease in female dogs. It usually occurs in middle age female dogs, usually about two months after the completion of heat. This disease is the accumulation of purulent fluid inside the uterus. Etiology of pyometra is not fully understood. It is assumed, that pyometra is a result of hormonal disorders in the endometrium combined with bacterial superinfection. The diagnosis is based on the interview, clinical examination, additional laboratory tests and ultrasound or x-ray of the abdomen. There are two treatments: ovariohysterectomy and conservative treatment with pharmacological agents for example prostaglandin, aglepriston, antibiotics with a broad spectrum of action. Currently conducted molecular studies have a large influence on the development of the present knowledge on the pathogenesis and course of pyometra, whose conclusions may be used to change the current therapeutic protocols.