ROMO1 is required for mitochondrial metabolism during preimplantation embryo development in pigs

Abstract

Background Reactive oxygen species (ROS) modulator 1 (ROMO1) is a mitochondrial membrane protein that is essential for the regulation of mitochondrial ROS production and redox sensing. ROMO1 regulates ROS generation within cells and is involved in cellular processes, such as cell proliferation, senescence, and death. Our purpose is to investigates the impact of ROMO1 on the mitochondria during porcine embryogenesis. Results We found that high expression of ROMO1 was associated with porcine preimplantation embryo development, indicating that ROMO1 may contribute to the progression of embryogenesis. Knockdown of ROMO1 disrupted porcine embryo development and blastocyst quality, thereby inducing ROS production and decreasing mitochondrial membrane potential. Knockdown of ROMO1 induced mitochondrial dysfunction by disrupting the balance of OPA1 isoforms to release cytochrome c, reduce ATP, and induce apoptosis. Meanwhile, ROMO1 overexpression showed similar effects as ROMO1 KD on the embryos. Overexpression of ROMO1 rescued the ROMO1 KD-induced defects in embryo development, mitochondrial fragmentation, and apoptosis. Conclusions ROMO1 plays a critical role in embryo development by regulating mitochondrial morphology, function, and apoptosis in pigs.

Oroxin A reduces oxidative stress, apoptosis, and autophagy and improves the developmental competence of porcine embryos in vitro

“…In this study, OA supplementation effectively improved the MMP of porcine early embryos, suggesting that OA can strengthen mitochondrial function. These findings are consistent with those of previous research, in which embryos with better mitochondrial function showed better proliferation and development(Niu et al, 2020; Zhou et al, 2021). .…”

“…In this study, OA supplementation effectively improved the MMP of porcine early embryos, suggesting that OA can strengthen mitochondrial function. These findings are consistent with those of previous research, in which embryos with better mitochondrial function showed better proliferation and development (Niu et al, 2020;Zhou et al, 2021).. In summary, this study suggested that supplementation of the IVC medium with 2.5 μM of OA could augment the quality of early embryos and enhance blastocyst rates and total cell numbers.…”

Epigallocatechin-3-gallate protects porcine oocytes against post-ovulatory aging through inhibition of oxidative stress

“…As previously report [ 23 ], a total of approximately 100 porcine oocytes per group were lysis with 10 μl RIPA buffer and 10 μl loading buffer, and then heated at 98 C for 10 min. Lysates were separated by 6–12% SDS-PAGE gel and transferred onto polyvinylidene fluoride membranes.…”

ATF7‐dependent epigenetic changes induced by high temperature during early porcine embryonic development

“…To prepare ATF7 dsRNA, ATF7 was amplified using a pair of primers containing the T7 promoter sequence ( ATF7 dsRNA: forward, TAA TAC GAC TCA CTA TAG GGG GGC TAT GAT CCA CTT CAC CC; reverse, TAA TAC GAC TCA CTA TAG GGG CTC ATC CGG ATC TTC ATC CA). The purified PCR products were used to synthesize dsRNA using the MEGAscript T7 Kit (AM1333; Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer's instructions 32,33 . After in vitro transcription, dsRNA was treated with DNase I and RNase A to remove the DNA template and any single‐stranded RNA, followed by purification by phenol–chloroform extraction and isopropyl alcohol precipitation.…”

“…The purified PCR products were used to synthesize dsRNA using the MEGAscript T7 Kit (AM1333; Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer's instructions. 32 , 33 After in vitro transcription, dsRNA was treated with DNase I and RNase A to remove the DNA template and any single‐stranded RNA, followed by purification by phenol–chloroform extraction and isopropyl alcohol precipitation. The purified dsRNA was dissolved in RNase‐free water.…”