Abstract
Oviductal epithelial cells (OECs) actively produce stimulating and protecting factors, favoring survival and viability of gametes and early embryos. The oviduct participates in the initial reproductive events, which strongly depends on adhesion. The analysis of differential gene expression in OECs, during long-term in vitro culture, enables recognition of new molecular markers regulating several processes, including “biological adhesion”. Porcine oviducts were stained with hematoxylin and eosin, as well as with antibodies against epithelial markers. Then, OECs were long-term in vitro cultured and after 24 h, 7, 15, and 30 days of culture were subjected to transcriptomic and proteomic assays. Microarrays were employed to evaluate gene expression, with Matrix-assisted laser desorption/ionization-time of light (MALDI-TOF) mass spectrometry applied to determine the proteome. The results revealed proper morphology of the oviducts and typical epithelial structure of OECs during the culture. From the set of differentially expressed genes (DEGs), we have selected the 130 that encoded proteins detected by MALDI-TOF MS analysis. From this gene pool, 18 significantly enriched gene ontology biological processes (GO BP) terms were extracted. Among them we focused on genes belonging to “biological adhesion” GO BP. It is suggested that increased expression of studied genes can be attributed to the process of intensive secretion of substances that exhibit favorable influence on oviductal environment, which prime gametes adhesion and viability, fertilization, and early embryo journey.
Highlights
The oviduct is a cylindrical structure extending between the ovary and the uterus
The most prominent changes in oviductal epithelial cells (OECs) isolated from the ampulla were related to genes associated with cell migration and motility, ciliary movement, and beat frequency
We only discuss genes that were differentially expressed during 30 days of OEC culture, protein products of which were found in cells on the last day of culture
Summary
The oviduct is a cylindrical structure extending between the ovary and the uterus. It is organized into three segments (the infundibulum, the ampulla, and the isthmus) and four histological layers (mucosa, submucosa, muscularis, and serosa). Transient adhesion of COCs to OECs is most likely the signal indicating the occurrence of ovulation, triggering sperm hyperactivation and release [10]. This passage is crucial for successful fertilization and ensures that only a low number of selected spermatozoa reaches the ampullar-isthmic junction where fertilization takes place [11]. EEaacchh ooff tthhee aannaallyyzzeedd sseeccttiioonnss sshhoowweedd nnoorrmmaall hhiissttoollooggiiccaall ssttrruuccttuurree. It was performed to indicate changes in Collagen Type I Alpha 1 Chain (COL1A1), Fibronectin 1 (FN1), and Talin 1 (TLN1) protein levels during OEC primary long-term in vitro culture (Figure 7) as genes encoding them presented the most substantial changes in RNA expression. An increase in FN1 protein level was more pronounced in 15 and 30 days of culture (~500%) as compared to those in 24 h
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