Analysis of fructose and mannose – regulatory peptides signaling pathway in porcine epithelial oviductal cells (OECs) primary cultured long-term in vitro

Abstract

Advances in Cell BiologyAnalysis of fructose and mannose -regulatory peptides signaling pathway in porcine epithelial oviductal cells (OECs) primary cultured long-term in vitro AbstractThe morphological and biochemical modification of oviductal epithelial cells (OECs) belongs to the compound process responsible for proper oocytes transport and successful fertilization. However, the main mechanisms which regulated this process are still not entirely known. Moreover, the OECs metabolism, which may be identified as the "cellular activity" marker, is poorly recognized. In this study we investigated the fructose and mannose metabolic pathway in porcine OECs primary long-term cultured in vitro. In our study, we employ a primary long term in vitro culture (IVC) and microarray approach (the Affymetrix microarray were used for analysis of transcriptomic profile of OECs) for expression levels analysis. We found that from the whole analyzed transcriptome, 1537 genes were upregulated and 995 were down regulated after 7 days of culture, 1471 genes were upregulated and 1061 were downregulated after 15 days of culture and 1329 genes were upregulated and 1203 were downregulated after 30 days of culture. Moreover, the differential expression of SORD, FPGT, PFKFB4, TPI1, MPI, ALDOC, HK2 and PFKFB3 at 24 hours, 7 day, 15 day and 30 day, was also observed. We suggested that fructose and mannose metabolism may be important molecular bio-marker of porcine OECs capability in in vitro model. The metabolic profile is significantly accompanied by cells proliferation in vitro. The transcriptomic profile of SORD, FPGT, PFKFB4, TPI1, MPI, ALDOC, HK2 and PFKFB3 expression may be identified as "fingerprint" of fructose and mannose metabolism in OECs as well as involved in cellular in vitro developmental capacity in pigs.Running title: Regulation of fructose and mannose metabolism

Nucleotide, ribonucleotide and ribonucleoside binding belongs to differentially expressed genes in porcine epithelial oviductal cells during longterm primary cultivation

“…Therefore, the goal of the present project is to describe molecular changes in OECs during primary in vitro long-term culture, through employing microarray approach. Previous studies have shown various aspects of OECs' morphological modifications and transcriptomic profile changes during in vitro cultivation [3][4][5]. The emphasis of the present study, is on groups of genes associated with nucleotide/ribonucleotide binding mechanism.…”

“…The results presented in previous studies are a valuable contribution for better understanding and elucidating many biochemical pathways and morphological/metabolomic factors existing in epithelial oviductal cells. Thus, employing a microarray approach, the molecular level processes identified were angiogenesis and blood vessels development [3], carbohydrates [4] and amino acids [5] metabolism. Furthermore, and plasticity of porcine cells associated with reproduction processes [6,7].…”

Ontology groups representing angiogenesis and blood vessels development are highly up-regulated during porcine oviductal epithelial cells long-term real-time proliferation – a primary cell culture approach

“…Epithelial oviductal cell lines may be a valuable tool for better understanding and elucidating many biochemical pathways existing in oviducts. Our previous studies have shown various aspects of OECs' morphological modifications and transcriptomic profile changes during in vitro cultivation [8,9]. Therefore, employing a microarray approach in our study, we aimed to investigate the transcriptomic profile of OECs during long-term in vitro primary cell culture.…”

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

“…For amplification, 2 μl of cDNA solution was added to 18 μl of QuantiTect ® SYBR ® Green PCR (Master Mix Qiagen GmbH, Hilden, Germany) and primers ( Table 1). One RNA sample of each preparation was processed without the RT-reaction to provide a negative control for subsequent PCR, where each sample processes in two technical repeats [17][18][19].…”