Melatonin improves the first cleavage of parthenogenetic embryos from vitrified–warmed mouse oocytes potentially by promoting cell cycle progression

Abstract

Background This study investigated the effect of melatonin (MT) on cell cycle (G1/S/G2/M) of parthenogenetic zygotes developed from vitrified-warmed mouse metaphase II (MII) oocytes and elucidated the potential mechanism of MT action in the first cleavage of embryos. Results After vitrification and warming, oocytes were parthenogenetically activated (PA) and in vitro cultured (IVC). Then the spindle morphology and chromosome segregation in oocytes, the maternal mRNA levels of genes including Miss, Doc1r, Setd2 and Ythdf2 in activated oocytes, pronuclear formation, the S phase duration in zygotes, mitochondrial function at G1 phase, reactive oxygen species (ROS) level at S phase, DNA damage at G2 phase, early apoptosis in 2-cell embryos, cleavage and blastocyst formation rates were evaluated. The results indicated that the vitrification/warming procedures led to following perturbations 1) spindle abnormalities and chromosome misalignment, alteration of maternal mRNAs and delay in pronucleus formation, 2) decreased mitochondrial membrane potential (MMP) and lower adenosine triphosphate (ATP) levels, increased ROS production and DNA damage, G1/S and S/G2 phase transition delay, and delayed first cleavage, and 3) increased early apoptosis and lower levels of cleavage and blastocyst formation. Our results further revealed that such negative impacts of oocyte cryopreservation could be alleviated by supplementation of warming, recovery, PA and IVC media with 10− 9 mol/L MT before the embryos moved into the 2-cell stage of development. Conclusions MT might promote cell cycle progression via regulation of MMP, ATP, ROS and maternal mRNA levels, potentially increasing the first cleavage of parthenogenetic zygotes developed from vitrified–warmed mouse oocytes and their subsequent development.

Melatonin Promotes in vitro Development of Vitrified-Warmed Mouse GV Oocytes, Potentially by Modulating Phosphorylation of Drp1

“…Melatonin regulated the level of p-Drp1 Ser616 to alleviate stress-induced damage, promoting cell development and functional recovery. Cryopreservation has been implicated in mitochondrial damage and reduced vitality (2,14,38,45). TOMM20 is a marker of mitochondrial (46,47).…”

“…The ratio of red fluorescence to green fluorescence was recorded as MMP ( ψm) of oocytes. The intensity of red and green fluorescence in each oocyte was measured using Image J software (Version 1.48; National Institutes of Health, Bethesda, MD, USA), calculated the fluorescence intensity was based on previous study (2,38,42).…”

“…It can also inhibit rotenone-induced SH-SY5Y cell death (33), maintain cardiac function in rats with mechanical trauma (23), protect cardiac microvasculature from ischemia/reperfusion injury (34) and treat prion disease (35) by regulating Drp1 activity. Furthermore, melatonin can reduce cryopreservation-induced mitochondrial damage, oxidative stress, and apoptosis in mouse (2,(36)(37)(38), horse (7), cattle (14) and human (39) oocytes. However, it remains to be explored whether melatonin involvement in the above-mentioned functions during in vitro development of vitrified-warmed mouse GV oocytes occurs via regulation of Drp1 or p-Drp1 Ser616.…”

Placental Dysfunction in Assisted Reproductive Pregnancies: Perinatal, Neonatal and Adult Life Outcomes

“…Melatonin upregulates the primary implantation receptors, ErbB1 and ErbB4, and significantly reduces intracellular ROS in mouse blastocysts (increasing the embryo total antioxidant capacity) and promotes mitosis of the inner cell mass and trophectoderm cells [ 58 ]. Recently, it has been demonstrated that the supplementation of culture medium with melatonin (10 −9 mol/L) improves the growth of mouse parthenogenetic embryo potentially by promoting cell cycle progression [ 59 ]. Despite all these functions, melatonin is not present in culture media used in ART.…”

Mitochondria Transfer from Adipose Stem Cells Improves the Developmental Potential of Cryopreserved Oocytes

“…This suggests that cryopreservation could damage the development of calcium transients at fertilization [ 18 ]. There have been many pre-clinical level research studies that mainly sought to improve mitochondrial biogenesis [ 19 ], decrease ROS production [ 20 ], and reestablish Ca 2+ ion regulation [ 21 ] in cryopreserved oocytes. Although these pre-clinical studies showed some improvement in embryo development, none have been able to restore the damaged oocyte mitochondria following cryopreservation.…”