Integrative DNA Methylation and Gene Expression Analyses Identify DNA Packaging and Epigenetic Regulatory Genes Associated with Low Motility Sperm

In previous studies using candidate gene approaches, low sperm count (oligospermia) has been associated with altered sperm mRNA content and DNA methylation in both imprinted and non-imprinted genes. We performed a genome-wide analysis of sperm DNA methylation and mRNA content to test for associations with sperm function. (NCBI 1788). There was a trend among altered expression of these epigenetic regulatory genes and RPMM DNA methylation class.Using integrative genome-wide approaches we identified CpG methylation profiles and mRNA alterations associated with low sperm motility

The in vitro culture supplements and selected aspects of canine oocytes maturation

The maturation of oocytes is one of the most important steps determining their developmental competence. Due to the low percentage of oocytes of bitches that reach the MII stage, searching for reagents that may stimulate the growth and maturation of oocytes is still present in this species of mammals. The most important media supplements include gonadotropins (LH, FSH, hCG), growth factors (IGF, TGF, EGF, FGF), progesterone and follicular fluid. It is suggested that the supplement of EGF, and/or follicular cells may have an important influence on the percentage of cells that reach the MII stage. Despite plenty of research based on the improvement of bitch oocytes in vitro culture, the results obtained are still unsatisfactory. Moreover, in the long stages of canine oocytes maturation many molecular and morphological modifications (including changes in mitochondria structure and configuration in the cytoplasm) are involved. In this article, the influence of selected media supplements on the efficiency of bitch oocytes in vitro maturation was described. The molecular and morphological modifications during canine oocytes maturation were also considered in the text

Analysis of integrins and vascular endothelial growth factor isoforms mRNA expression in the canine uterus during perimplantation period

Integrins are the major receptors within the extracellular matrix (ECM) that mediate several functions connected with cell life and metabolism, such as cell adhesion, migration, cytoskeletal organization, proliferation, survival, and differentiation. A vascular endothelial growth factor (VEGF) is one of the most important angiogenic factors. It has been suggested that the expression of this gene may play crucial physiological roles in reproductive organs. All investigated endometrial tissues were isolated on day 10-12 after mating. Control bitches, used in this study, were in metestrus, which was determined according to the vaginal cytology and progesterone level in blood. Early pregnancy was verified by flushing the uterine horns with PBS. Total RNA was isolated from the bitches endometrium by means of the Chomczyński and Sacchi method, treated by DNase I, and reverse-transcribed into cDNA. A quantitative analysis of integrins α2b, β2 and β3, VEGF 164, 182 and 188 cDNA was performed by RT-PCR. In results we have shown an increased expression of all investigated genes (integrins α2b, β2 and β3, VEGF 164, 182, and 188) in pregnant bitches uterus as compared to non-pregnant females (P<0.001). Our results indicated that the expression of genes encoding integrins and vascular endothelial growth factors is different in relation to the time of the embryo implantation and it is increased in the first period of this process. This may be associated with the induction of specific mechanisms responsible for receptivity of uterus following the embryo attachment. In addition, all of investigated genes are up-regulated in a pregnancy-specific manner and the increased expression of these genes may regulate the uterus function during the implantation of canine embryos

Differential expression of epidermal growth factor and transforming growth factor beta isoforms in dog endometrium during different periods of the estrus cycle

Both epidermal growth factor (EGF) and transforming growth factor (TGF) play an important physiological role in the processes of proliferation and differentiation of several different cell types. However, the expression profiles of these factors in domestic bitches endometrium are still poorly recognized. The aim of the present study was to identify and analyze the differential expression of these factors in various stages of the estrus cycle. Endometrial tissue from proestrus (n=17), estrus (n=10), day 10 diestrus (n=15), day 35 diestrus (n=18) and anestrus (n=25) was collected soon after ovariohysterectomy. Total RNA was isolated from the endometrium by means of Chomczyński and Sacchi method, treated by DNase I, and reverse- transcribed into cDNA. Quantitative analysis of EGF, TGFβ1, TGFβ2, and TGFβ3 cDNA was performed by real-time quantitative polymerase chain reaction (RT-PCR). EGF expression in canine endometrium was increased in the estrus stage as compared to proestrus (P<0.05), day 10 diestrus (P<0.05), day 35 diestrus (P<0.01) and anestrus (P<0.001). We also found the differences in EGF expression between day 10 and day 35 of estrus as well as between day 35 of estrus with anestrus (P<0.05, P<0.01, respectively). The TGFf1 transcript contents were also higher in estrus as compared to other stages (P<0.01). The TGFβ2 and TGFβ3 in the estrus stage was increased compared to proestrus, day 10 diestrus, day 35 diestrus and anestrus (P<0.05). We proved that expression of EGF and TGFβ transcript isoforms is related to the phase of estrus in bitches and therefore may be regulated by specific hormone concentrations during these periods. Our results confirm the hypothesis that these growth factors play a role in the regulation of biochemical changes in the endometrial tissues during the estrus cycle

Salinity gradient energy conversion by custom-made interpolymer ion exchange membranes utilized in reverse electrodialysis system

Reverse electrodialysis (RED) is one of methods to extract salinity gradient energy between two aqueous solutions with different salt concentrations. In this work, custom-made interpolymer ion exchange membranes were employed in the RED stack. The effects of divalent (Mg2+, Ca2+ and SO42-) and monovalent (Li+, K+ and Cl-) ions in the feed solutions prepared from NaCl salt as a function of such process parameters as number of membrane pairs, flow rate, and salinity ratio on power generation by the RED method were studied. It was shown that the maximum power density of 0.561 W/m2 was reached by using three membrane pairs using 1:45 of salt ratio with a feed flow rate of 120 mL/min using only NaCl salt in the feed solutions. The maximum power density was 0.398 W/m2 at 120 mL/min of the flow rate of the feed solutions composed of 90 wt% NaCl and 10 wt% KCl by using a salt ratio of 1:30 while the lowest power density of 0.246 W/m2 was obtained with a feed flow rate of 30 mL/min in the presence of SO42- ions with a similar salt ratio. Consequently, it was seen that while the presence of divalent ions in NaCl solutions had negative impact on power generation by RED system, the addition of monovalent ions having smaller hydrated radius than that of the Na+ ions contributed positively to the power generation. © 2023 Elsevier Ltd.This research was funded by TÜBİTAK , grant number TÜBİTAK 117M023 .YÖK 100-2000; Türkiye Bilimsel ve Teknolojik Araştırma Kurumu, TÜBİTAK: 117M023This research has been financially supported by the bilateral collaboration programme (TUBITAK-NCBR-2549) between Turkey and Poland (Project No: TÜBİTAK 117M023). E. Altıok is grateful for the Ph.D. scholarship of Turkish Higher Education Council ( YÖK 100-2000 )