Interaction of the EGFR signaling pathway with porcine cumulus oocyte complexes and oviduct cells in a coculture system.

It has become increasingly recognized that coculture has a beneficial effect on the in vitro maturation (IVM) of oocytes and embryo development in many species. However, these effects of coculture on IVM have been documented only for their positive conditioning roles without any evidence on the precise mechanisms underlying the action of coculture systems on the development of cumulus oocyte complexes (COCs). It has been suggested that the epidermal growth factor receptor (EGFR) signaling pathway is important for development of COCs, mediated by several epidermal growth factor (EGF)-like proteins with downstream mitogen-activated protein kinase 1/3 signaling. Therefore, we hypothesized that canine oviduct cells (OCs) in a coculture system, which shows improvement of oocyte quality in several species, are associated with EGFR signaling by exposure to progesterone (P4; imitating its production before ovulation and its continuous increase while oocytes reside in the oviduct to complete maturation in dogs). We designed three experimental groups: control, OCs coculture exposed to P4, and OCs coculture without exposure to P4. The result showed that the OCs coculture exposed to P4 strongly expressed EGF-like proteins and significantly improved COCs and subsequent embryo development. Furthermore, the expression of EGFR-related genes in cumulus cells and GDF9 and BMP15 in oocytes was upregulated in the P4-treated group. This study provides the first evidence that OCs exposed to P4 can induce strong expression of EGF-like proteins, and OCs effectively mediate improved porcine COCs development and subsequent embryo development by altering EGFR signaling related mRNA expression.

Lee SH ，Oh HJ ，Kim MJ ，Setyawan EMN ，Lee BC ... -  《-》

Effect of co-culture canine cumulus and oviduct cells with porcine oocytes during maturation and subsequent embryo development of parthenotes in vitro.

In the estrus stage, canine oocytes are surrounded by cumulus cells and undergo maturation in the oviduct for 2-3 days after ovulation. We hypothesized that canine oviduct cells (cOC) and canine cumulus cells (cCC) during this stage might affect the maturation of oocytes and thereby improve subsequent embryo development. Therefore, the objective of this study was to compare the effects of a cOC and cCC co-culture on oocyte in vitro maturation (IVM) and subsequent embryo development, and to analyze the gene expressions in a molecular fashion what co-culture actually gives the specific pathways in which the co-culture cells act to improve maturation and embryo development. The effect of co-culture using cOC and cCC on porcine oocyte IVM was investigated. Thereafter, oocytes were activated using electrical stimulation and embryo developmental competence was estimated. The expression of the genes related to oocyte maturation, embryo development and apoptosis were analyzed. Also, reactive oxygen species (ROS) levels after IVM was analyzed. The IVM rate and embryo development including cleavage, blastocyst formation rates, and total blastocyst cell numbers from cOC group were significantly higher than other groups (P < 0.05). The expression of SMAD2/3 and growth differentiation factor 9 (GDF9) was significantly increased in cOC and oocytes from the cOC group compared with other groups. Moreover, the levels of GDF9, prostaglandin-endoperoxide synthase 2 (PTGS2), WNT3A and matrix metalloproteinase 2 (MMP2) were significantly up-regulated in blastocysts from the cOC group. The concentration of ROS was significantly lower in the supernatant of cOC groups compared with other groups. Also, the expression of BCL2 was significantly increased in porcine cumulus cells and oocytes from cOC group. The present study demonstrated that co-culture with cOC improved in vitro porcine oocyte maturation and subsequent embryo development competence. Also, co-culture with cOC during IVM induces a suitable environment for oocyte maturation by enhancing the mRNA level of SMAD2/3 and GDF9, and for embryo development by elevating the expression level of PTGS2, WNT3A and MMP2. In addition, the decreased ROS level in cOC co-culture could have a beneficial influence on oocyte maturation.

Lee SH ，Oh HJ ，Kim MJ ，Kim GA ，Choi YB ，Jo YK ，Setyawan EMN ，Lee BC ... -  《-》

Exosomes derived from oviduct cells mediate the EGFR/MAPK signaling pathway in cumulus cells.

Current studies indicate that application of oviduct cells (OCs) in in vitro system create microenvironment similar to the in vivo conditions by releasing multiple growth factors which has beneficial effects on the development of cumulus-oocyte complexes and embryos. In particular, recent evidence with a coculture system indicates that there is a reciprocal relationship between canine OCs and cumulus cells and that oviductal secretions can promote changes in cellular protein/gene expression. Despite the fact that OCs respond to cumulus cells, a clear understanding of the mechanism by which the components released from OCs that play a role in modulating the biological function of cumulus cells is still elusive. Therefore, we hypothesized that exosomes derived from OCs (OC-Exo), which efficiently mediate cellular communication by transferring their molecular cargo to recipient cells, could be key modulators of the cross-talk with cumulus cells. We aimed to characterize OC-Exo and decipher their physiological effects on cumulus cells via the epidermal growth factor receptor/mitogen-activated protein kinase (EGFR/MAPK) pathway, which is one of the prerequisite pathways for cell development. Exposure of OC-Exo improved physiological cumulus cell condition including cell concentration, viability, and proliferation rate could reduce the accumulation of reactive oxygen species and the apoptotic rate. Moreover, exosomes could enhance the messenger RNA transcript and protein levels related to EGFR signaling in cumulus cells. The present study provides the first evidence that OC-Exo effectively enhance the physiological condition of cumulus cells exposed to GW4869 or Gefitinib via the EGFR/MAPK signaling pathway and this could be the primary mediators of molecular interactions among cumulus cells and shedding light on the role of exosomes in cumulus cells might permit improvement of oocyte and embryo development in vitro.

Lee SH ，Oh HJ ，Kim MJ ，Lee BC ... -  《-》

Mouse oocytes enable LH-induced maturation of the cumulus-oocyte complex via promoting EGF receptor-dependent signaling.

Su YQ ，Sugiura K ，Li Q ，Wigglesworth K ，Matzuk MM ，Eppig JJ ... -  《-》

Exogenous Ganglioside GT1b Enhances Porcine Oocyte Maturation, Including the Cumulus Cell Expansion and Activation of EGFR and ERK1/2 Signaling.

Kim JW ，Park HJ ，Yang SG ，Kim MJ ，Kim IS ，Jegal HG ，Wee G ，Yang HY ，Park JJ ，Choo YK ，Koo DB ... -  《-》