Adiponectin Improves In Vitro Development of Cloned Porcine Embryos by Reducing Endoplasmic Reticulum Stress and Apoptosis

Abstract

The main factor of embryonic demise is endoplasmic reticulum (ER) stress. Successful attenuation of ER stress results in an improvement in embryo development. We studied the impact of adiponectin in the in vitro culture (IVC) of porcine embryos derived from parthenogenetic activation and somatic cell nuclear transfer (SCNT). The first experiment revealed that 15 and 30 μg/mL adiponectin treatments improved cleavage, blastocyst rates, and total cell number (TCN) of parthenogenetic embryos and reduced the expression of XBP1 compared to the 5 μg/mL adiponectin treatment and control groups (p < 0.05). The second experiment showed that cleavage rate, blastocyst formation rate, and TCN of blastocysts were improved in the 15 μg/mL adiponectin treatment group compared with the control group, with significantly reduced XBP1 expression in ≥4-cell stage SCNT embryos and blastocysts (p < 0.05). Treatment with 15 μg/mL adiponectin significantly improved the expression of XBP1 and reduced the expression of ER stress-related genes (uXBP1, sXBP1, PTPN1, and ATF4), increased the expression levels of pluripotency-related genes (Nanog and SOX2), and decreased apoptosis-related gene expression (Caspase-3). These results suggest that 15 μg/mL adiponectin enhanced the in vitro developmental capacity of early-stage SCNT porcine embryos by reducing ER stress and apoptosis.

Strategies to Improve the Efficiency of Somatic Cell Nuclear Transfer

“…In addition, adiponectin (a protein hormone and adipokine) supplementation in IVC media significantly increased cleavage and blastocyst rates as well as the total cell number of cloned porcine embryos. Adiponectin reduced the level of XBP1 expression and ER stress-related genes, enhanced the expression levels of NANOG and SOX2, and decreased that of Caspase-3 [210].…”

Porcine oviductal extracellular vesicles facilitate early embryonic development via relief of endoplasmic reticulum stress

“…XBP1, an essential activator for regulating the gene expression of UPR signaling during ER stress, is generally used as an ER stress marker both in vivo and in vitro (Lin et al, 2019; Zhang, Diao, Oqani, et al, 2012). Inhibiting the expression of active XBP1 was found to improve early embryonic development by relieving ER stress (Ridlo et al, 2021; Zhang, Diao, Kim, et al, 2012). As such, we speculated that the presence of oviductal EVs should ameliorate ER stress in preimplantation embryos.…”

iSCNT embryo culture system for restoration of Cervus nippon hortulorum, presumed to be sika deer in the Korean Peninsula

“…In this context, an important role is also played by the strategies used to artificially synchronize the mitotic cycle of ex vivo-expanded nuclear donor cells at the G0/G1 stages [ 8 – 10 ]. It is also noteworthy that the developmental outcome of somatic cell-cloned embryos is largely determined by the molecular quality parameters reflected in the incidence of apoptotic cell death and oxidative stress processes in the in vitro cultured nuclear donor cells and SCNT-derived embryos [ 11 , 12 ]. Moreover, the developmental capability of cloned embryos is remarkably affected by the molecular quality of metaphase II-stage nuclear recipient oocytes, which depends largely on coordination between the processes of meiotic, cytoplasmic, and epigenomic maturation [ 13 – 15 ].…”