Recent results indicate a key role for cyclic guanosine monophosphate (cGMP) in the regulation of oocyte meiotic arrest preovulatory mammalian follicles. The aim our study was to determine whether resumption meiosis and expansion cumulus cells isolated pig cumulus-oocyte complexes (COCs) can be blocked by high intracellular concentration cGMP, this effect is mediated cGMP-dependent inhibition mitogen-activated protein kinase 3/1 (MAPK3/1).The COCs were from ovaries slaughtered gilts cultured...

The aim of this work was to assess the FSH-stimulated expression epidermal growth factor (EGF)-like peptides in cultured cumulus–oocyte complexes (COCs) and find out effect on cumulus expansion, oocyte maturation, acquisition developmental competence vitro . FSH promptly stimulated amphiregulin ( AREG ) epiregulin EREG ), but not betacellulin BTC COCs. Expression reached maximum at 2 or 4 h after addition respectively. also significantly expansion-related genes PTGS2 , TNFAIP6 HAS2 COCs 8...

Abstract All porcine oocytes cultured 20 hr in medium with 10 μg/ml cycloheximide rested the germinal vesicle (GV) stage but highly condensed bivalents nucleoplasm. When these were washed and control for 2, 4, 6 hr, breakdown (GVBD) was completed 0, 86, 100% of them, respectively. similarly inhibited successively only 2.5 given again enriched (3.5 hr), nearly all them reached late diakinesis again. It means that free this inhibitor protein synthesis GVBD rapidly (4 hr) crucial nuclear...

To define signaling pathways that drive FSH- and epidermal growth factor (EGF)-like peptide-induced cumulus expansion oocyte meiotic resumption, in vitro cultured pig cumulus-oocyte complexes were treated with specific protein kinase inhibitors. We found FSH-induced maturation of oocytes was blocked germinal vesicle (GV) stage by A (PKA), MAPK14, MAPK3/1, EGF receptor (EGFR) tyrosine inhibitors (H89, SB203580, U0126, AG1478 respectively) whereas phosphoinositide-3-kinase/v-akt murine thymoma...

We have recently shown that epidermal growth factor (EGF) strongly stimulates expansion of porcine oocyte-cumulus complexes (OCCs) isolated from large follicles (>6 mm) and does not promote OCCs small (3-4-mm) follicles. In order to elucidate the role EGF in expansion, present study, we first examined presence receptors (EGFRs) cumulus cells different sizes. Surprisingly, immunoblotting showed obtained all follicular size categories contained similar amounts EGFR protein. On other hand,...

Summary. Oocyte–cumulus cell complexes (OCC) and with an attached piece of membrana granulosa (C + P), isolated from prepubertal or cyclic gilts stimulated pregnant mares' serum gonadotrophin, were cultured in media supplemented follicle-stimulating hormone (FSH; 0·01–1·0 μg/ml) forskolin (50–100 μmol/l) for 24 32 h. FSH each induced dose-dependent cumulus expansion. After 2 h culture, (0·1 (100 increased the contents intracellular adenosine 3′,5′-phosphate (cAMP) OCC to almost 10 times that...

The gonadotropin-induced resumption of oocyte meiosis in preovulatory follicles is preceded by expression epidermal growth factor (EGF)-like peptides, amphiregulin (AREG) and epiregulin (EREG), mural granulosa cumulus cells. Both the gonadotropins EGF-like peptides possess capacity to stimulate vitro via activation a broad signaling network To better understand rapid genomic actions (FSH) we analyzed transcriptomes cells at 3 h after their stimulation.We hybridized aRNA from pig...

Abstract Elevated summer temperature is reported to be the leading cause of stress in dairy and beef cows, which negatively affects various reproductive functions. Follicular cells respond heat (HS) by activating expression shock family proteins (HSPs) other antioxidants. HS affect bi-directional communication between follicular oocyte, partly mediated fluid extracellular vesicles (EVs) released from surrounding cells. As carriers bioactive molecules (DNA, RNA, protein, lipids), involvement...

We have shown previously that porcine cumulus and mural granulosa cells produce a factor is very similar, if not identical, to the oocyte-derived expansion-enabling (CEEF). Because growth differentiation 9 (GDF9) most likely candidate for CEEF, in present study we tested hypothesis GDF9 expressed only oocytes pig but also somatic follicular cells. In addition, asked whether relative abundance (RA) of mRNA changes and/or during periovulatory period or culture oocyte-cumulus complexes (OCCs)...

In recent studies using a rat aortic balloon occlusion model, we have demonstrated that spinal grafting of or human neuronal precursors postmitotic hNT neurons leads to progressive amelioration spasticity and rigidity corresponding improvement in ambulatory function. the present study, characterized optimal dosing regimen safety profile stem cells (HSSC) when grafted into lumbar cord segments naive immunosuppressed minipigs. Gottingen-Minnesota minipigs (18–23 kg) were anesthetized with...

Abstract Innumerable similarities in reproductive cyclicity and hormonal alterations highlight the considerable utility of mare to study aspects follicular dynamics function view largely constricted, human research subjects. The bi-directional communication between growing oocyte surrounding somatic cells embodies hallmark mammalian development, partially mediated by extracellular vesicles (EVs) encapsulated with microRNAs (miRNAs) present fluid (FF). Here, we aimed decipher miRNAs EVs from...

Abstract Background Oocytes of large animal species isolated from small ovarian follicles (< 2 mm) are less competent to support early embryonic development after in vitro maturation and fertilization than their counterparts medium-sized preovulatory follicles. This study aimed assess the effect a new medium containing FGF2, LIF, IGF1 (FLI medium) on meiotic developmental competence pig cumulus-oocytes complexes (COCs) derived Methods The growing oocytes were 1 (small follicle; SF)...

Abstract In this article, we focused on the impact of precisely chemically modified FLI maturation medium enriched with fibroblast growth factor 2 (FGF2), leukemia inhibitory (LIF), insulin-like 1 (IGF1), and polyvinyl alcohol (PVA) its potential to improve efficiency in vitro production porcine embryos. We hypothesized that enhancing composition could result an elevated embryos can affect EGA . resulted a significantly higher rate oocyte blastocyst formation compared control DMEM medium....

Previous studies have shown that the heavy chains (HCs) of serum-derived inter-alpha-trypsin inhibitor (IalphaI) molecules become covalently linked to hyaluronan (HA) during in vivo mouse cumulus expansion and significantly contribute matrix organization. Experiments with mice suggest incorporation such proteins appears be rather complex, involving LH/hCG-induced changes blood-follicle barrier functional cooperation between cells, granulosa oocyte within follicle. We demonstrate here HC-HA...

The purpose of the present study was to elucidate signaling pathways by which insulin like-growth factor 1 (IGF1) promotes FSH-stimulated synthesis and retention hyaluronic acid (HA) in pig oocyte-cumulus complexes (OCCs) cultured serum-free medium. We found that IGF1 had no effects on production cAMP activation protein kinase A OCCs. Immunoblotting with phospho-specific antibodies showed FSH moderately phosphorylated v-akt murine thymoma viral oncogene homolog (AKT) mitogen-activated 3...

Several lines of evidence suggest that in mice the activation SMAD2/3 signaling by oocyte secreted factors, together with epidermal growth factor receptor (EGFR) activation, is essential to induce cumulus expansion. Here we show inhibition EGFR kinase follicle stimulating hormone (FSH)-stimulated porcine oocyte-cumulus cell complex (OCCs) strongly decreases hyaluronan (HA) synthesis and its retention matrix, as well progesterone synthesis. Although cells undergo expansion independently...

The developmental potential of porcine oocytes cultured in vitro was remarkably enhanced a medium containing FGF2, LIF and IGF1 (FLI) when compared to supplemented with gonadotropins EGF (control). We analyzed the molecular background oocyte quality by comparing time course MAPK3/1 AKT activation, expression genes controlled these kinases cumulus-oocyte complexes (COCs) FLI control medium. pattern activation COCs very similar both media, except for robust increase phosphorylation during...

Ovarian follicular fluids (FFs) contain several kinds of regulatory factors that maintain a suitable microenvironment for oocyte development. Extracellular vesicles (EVs) are among the play essential roles in regulating follicle and development through their cargo molecules include microRNAs (miRNAs). This study aimed to investigate small-EV (s-EV) miRNAs porcine FFs potential association with quality.Individual aspirated oocytes were stained lissamine green B stain (LB), vital quality, each...

Abstract Oocyte developmental competence is acquired during folliculogenesis and regulated by complex molecular mechanisms. Several molecules are involved in these mechanisms, including microRNAs (miRNAs) that essential for oocyte‐specific processes throughout the development. The objective of this study was to identify expression profile miRNAs porcine oocytes derived from follicles different sizes using RNA deep sequencing. Oocytes were aspirated large (LO; 3–6 mm) or small (SO; 1.5–1.9...

Mouse oocytes secrete a factor that enables cumulus cells to undergo expansion in response FSH (1 μg/ml), whereas of the porcine oophorus has been shown be independent oocyte. The aim this study was assess FSH-induced synthesis hyaluronic acid (HA) by before and after oocytectomy. In addition, we studied effect insulin-like growth factor-I (IGF-I) on ability synthesize retain HA serum-free medium. Porcine oocyte-cumulus complexes from which had removed oocytectomy were cultured for 24 h...

The objective of this study was to find out whether porcine cumulus and mural granulosa cells can secrete expansion-enabling factor (CEEF). Culture drops M-199 medium were conditioned with denuded oocytes (1 oocyte/μl), from oocytectomized complexes OOX/μl), pieces isolated preantral preovulatory follicles (1000 cells/μl), or oviductal cells/μl) for 24 hr. production CEEF assessed by the addition mouse OOX follicle-stimulating hormone (FSH) μg/ml) microdrops medium. After 16–18 hr, expansion...