Effects of Addition of Tissue‐Type Plasminogen Activator in In Vitro Fertilization Medium on Bovine Embryo Development and Quality

Abstract

Plasminogen activators/Plasmin system plays pivotal role in regulating reproductive functions of mammals. Here, we examined the effects of modification of in vitro fertilization medium (IVF medium) with the addition of tissue-type plasminogen activator (t-PA), on bovine embryo development and quality, assessed by quantification of expression of various genes related to metabolism, oxidation, implantation and apoptosis. In addition, plasminogen activator activity (PAA) and plasminogen activator inhibition (PAI) were measured in the spent media. After conventional IVM, 2016 cumulus-oocyte complexes (COCs) were divided into four groups with modified composition of the IVF medium containing t-PA and/or its inhibitor epsilon-aminocaproic acid (control, t-PA, t-PA+ε-ACA, ε-ACA). Presumptive zygotes were cultured for 8 days in synthetic oviductal fluid (SOF) medium; gene expression studies were carried out on morulae and blastocysts. t-PA alone significantly suppressed cleavage and blastocyst formation rates, but this effect was neutralized by the addition of ε-ACA. PAA in the treated group was significantly reduced by ε-ACA, but without total elimination. Significant differences were detected in the expression of genes related to apoptosis and/or cell cycle arrest (BAX, BCL2L1, KAT2B) between embryos produced in t-PA-modified media and controls, giving an overall notion that the inferior developmental competence of treated embryos may be attributed to apoptotic phenomena induced by t-PA. In conclusion, it appears that excessive t-PA content in the IVF media, suppresses blastocyst formation rate, possibly due to induction of apoptotic phenomena.

Urokinase-type plasminogen activator does not affect in vitro bovine embryo development and quality

“…Lastly, PLG and PLAUR mRNA expression were selected to investigate whether u-PA addition altered the relative mRNA abundance of those related genes which are members of the PAs/PL system. PLG mRNA expression was not detected either in control or in u-PA treated embryos, which is in accordance with our previous results for PLG expression in control day 7 embryos (Krania et al, 2015).…”

“…Acta Veterinaria Hungarica 63, 2015 Experiment 2: Effect of u-PA addition in IVC, on embryos cultured on a cumulus cell monolayer under serum-containing conditions Bovine embryos were produced as previously described (Dovolou et al, 2014;Krania et al, 2015).…”

miRNA profiles in livers with different mass deficits after partial hepatectomy and miR-106b~25 cluster accelerating hepatocyte proliferation in rats

“…It interacts with E2F1 and represses its transcriptional activity, thereby resulting in cell cycle arrest 27 . KAT2B inhibits cell cycle progression and counteracts the mitogenic activity of the adenoviral oncoprotein E1A 28 29 . Thus, it is conceivable that up-regulation of the miR106b~25 cluster during PH is vital for the reentry of quiescent hepatocytes into the cell cycle for regrowth.…”

Effects of 17β-estradiol, Interleukin-1β, and Human Chorionic Gonadotropin on Activity and mRNA Expression of Plasminogen Activators in Porcine Endometrial Cells

“…They play a role in reproductive and physiological processes including angiogenesis, oocyte maturation, embryo development, ovulation, activation of matrix metalloproteinases (MMPs), and degradation of collagens and ECM proteins ( Ebisch et al, 2008 ). In particular, sperm-zona pellucida binding is decreased by both PAs in the cytoplasm and zona pellucida of porcine oocytes during fertilization ( Coy et al, 2012 ), and Krania et al (2015) reported that addition of tPA during the in vitro fertilization of bovine oocytes decreased embryo development and increased the expression of apoptosis-related genes in embryos. Therefore, regulatory mechanisms underlying PA mRNA expression and activation in the female reproductive tract are important for animal reproduction.…”