Amino acids metabolism and degradation is regulated during porcine oviductal epithelial cells (OECs) primary culture in vitro – a signaling pathways activation approach

Abstract

The ovary is part of the reproductive system, possessing very important functions in the reproduction process (ovum and embryo transfer, providing a suitable environment for sperm capacitation, etc.). There are two types of cells in the fallopian tubes: alveolar and secretive cells. These study shows the metabolic processes in pig oviductal epithelial cells associated with the activation of signaling pathways of amino acids metabolism and degradation during long-term in vitro culture. Oviductal epithelial cells from 45 colonies in the anestrous phase of the estrous cycle have been utilized in this study. RNA extract from the OEC primary cultures was pooled after 24h, 7days, 15 days and 30 days from the beginning of culture and the transcriptome investigated by Affymetrix® Porcine Gene 1.1 ST. From the whole transcript that consisted of 2009 different genes, 1537 were upregulated and 995 were downregulated after 7 days of culture, 1471 were upregulated and 1061 were downregulated after 15 days of culture and 1329 were upregulated and 1203 were downregulated after 30 days of culture. The results of these studies provide, for the first time, information on the activation of metabolic pathways of amino acids such as valine, leucine, isoleucine, cysteine, and methionine in the investigated tissue. They also indicate genes that may be OECs-specific genetic markers that are expressed or upregulated during long-term in vitro culture.Running title: regulation of amino acid signaling pathways

Genes Involved in the Processes of Cell Proliferation, Migration, Adhesion, and Tissue Development as New Potential Markers of Porcine Granulosa Cellular ProcessesIn Vitro: A Microarray Approach

“…The Affymetrix procedure was performed as previously described (Kranc et al, 2017(Kranc et al, , 2018a(Kranc et al, , 2018bBryja et al, 2018;Chamier-Gliszczyńska et al, 2018;Stefańska et al, 2018;Nawrocki et al, 2018). cDNA was reverse transcribed from total sample RNA (100 ng) (Ambion Ò WT Expression Kit).…”

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].…”

Genes regulating biochemical pathways of oxygen metabolism in porcine oviductal epithelial cells during long-term IVC

“…The method used for OEC harvesting and culture has been described in our previous studies [17][18][19]. Briefly, collected oviducts were washed twice in Dulbecco's phosphate buffered saline (PBS) and sectioned longitudinally.…”