Treatment of allicin improves maturation of immature oocytes and subsequent developmental ability of preimplantation embryos

Zygote ◽  
2017 ◽  
Vol 25 (4) ◽  
pp. 480-488 ◽  
Author(s):  
Sang-Gi Jeong ◽  
Seung-Eun Lee ◽  
Yun-Gwi Park ◽  
Yeo-Jin Son ◽  
Min-Young Shin ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Polar Body ◽  
Treated Group ◽  
Developmental Competence ◽  
Mitogen Activated Protein Kinase ◽  
Preimplantation Embryos ◽  
Maturation Medium ◽  
Mitogen Activated Protein ◽  
Porcine Oocyte

SummaryAllicin (AL) regulates the cellular redox, proliferation, viability, and cell cycle of different cells against extracellular-derived stress. This study investigated the effects of allicin treatment on porcine oocyte maturation and developmental competence. Porcine oocytes were cultured in medium supplemented with 0 (control), 0.01, 0.1, 1, 10 or 100 μM AL, respectively, during in vitro maturation (IVM). The rate of polar body emission was higher in the 0.1 AL-treated group (74.5% ± 2.3%) than in the control (68.0% ± 2.6%) (P < 0.1). After parthenogenetic activation, the rates of cleavage and blastocyst formation were significantly higher in the 0.1 AL-treated group than in the control (P < 0.05). The reactive oxygen species level at metaphase II did not significantly differ among all groups. In matured oocytes, the expression of both BAK and CASP3, and BIRC5 was significantly lower and higher, respectively, in the 0.1 AL-treated group than in the control. Similarly, the expression of BMP15 and CCNB1, and the activity of phospho-p44/42 mitogen-activated protein kinase (MAPK), significantly increased. These results indicate that supplementation of oocyte maturation medium with allicin during IVM improves the maturation of oocytes and the subsequent developmental competence of porcine oocytes.

scholarly journals The effect of vitrification after warming on the expressions of p38, CDK1, and cyclin B in immature goat oocytes followed by in vitro maturation

2020 ◽  
Vol 13 (10) ◽  
pp. 2126-2132
Author(s):  
A. A. Muhammad Nur Kasman ◽  
Budi Santoso ◽  
Widjiati Widjiati
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Cyclin B ◽  
Mitogen Activated Protein Kinase ◽  
Oocyte Competence ◽  
Maturation Medium ◽  
Mitogen Activated Protein ◽  
Oocyte Meiotic Maturation

Background and Aim: The combination of vitrification techniques and in vitro maturation can reduce oocyte competence. Mitogen-activated protein kinase and maturation-promoting factor are significant in oocyte meiotic maturation regulation. This study aimed to analyze vitrification's effect, after warming followed by in vitro maturation, on the expressions of protein 38 (p38), cyclin-dependent kinase 1 (CDK1), and cyclin B and oocyte maturation level. Materials and Methods: Immature goat oocytes were soaked in vitrification and warming solutions. The procedure was followed by in vitro maturation and in vitro maturation without post-warming vitrification as a control. These oocytes, along with their cumulus, were vitrified using hemistraw in liquid nitrogen. Oocyte maturation was carried out in a maturation medium that was added with 10 μg/mL of FSH, 10 μg/mL of LH, and 1 μg/mL E2 for 22 h. The expressions of p38, CDK1, and cyclin B were observed using immunocytochemical methods, which were assessed semiquantitatively according to the modified Remmele method. The oocyte maturation level was observed using the aceto-orcein staining method based on the achievement of chromosomes up to the metaphase II stage and/or the formation of the polar body I. Results: p38 expression in vitrified oocytes after warming, followed by in vitro maturation, increased insignificantly (p≥0.05), with the acquisition of 3.91±2.69 and 2.69±0.50 in the control oocytes. CDK1 expression in vitrified oocytes decreased significantly (p≤0.05) after warming, followed by in vitro maturation, with the acquisition of 2.73±1.24 and 7.27±4.39 in the control oocytes. Cyclin B expression in vitrified oocytes decreased insignificantly (p≥0.05) after warming, followed by in vitro maturation, with the acquisition of 3.09±1.4 and 4.18±2.61 in the control oocytes. The proportion of vitrified oocyte maturation levels after warming, followed by in vitro maturation, decreased significantly (p≤0.05), with the acquisition of 45.45% and 77.27% in the control oocytes. Conclusion: This study concluded that vitrification after warming resulted in an insignificant increase in p38 expression, a significant decrease in CDK1 expression, an insignificant decrease in cyclin B expression, and a significant reduction in oocyte maturation levels.

185 PGI2 ANALOG INDUCES HIGH-QUALITY PORCINE BLASTOCYSTS IN VITRO

2007 ◽  
Vol 19 (1) ◽  
pp. 209 ◽  
Author(s):  
J.-S. Kim ◽  
G. Wee ◽  
B.-S. Song ◽  
J.-S. Park ◽  
X.-L. Jin ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Treated Group ◽  
Blastocyst Stage ◽  
Early Embryo ◽  
Mitogen Activated Protein Kinase ◽  
Blastocyst Formation ◽  
Prostaglandin I2 ◽  
Western Blots ◽  
Porcine Oocyte

Prostaglandin I2 (PGI2) exists in oviductal fluid and is synthesized from arachidonic acid by prostacyclin synthetase. In hatched embryos, prostaglandin I2 (PGI2) is related to implantation improvement, but its role during oocyte maturation and early embryo development remains controversial. Therefore, in this study, the effect of addition of a PGI2 analog during early porcine oocyte maturation on nuclear maturation, blastocyst formation, and pre-implantation embryonic quality was investigated. Porcine oocytes were matured in NCSU-23 medium supplemented with 10% (v/v) porcine follicular fluid, 10 ng mL-1 epidermal growth factor, 25 �M �-mercaptoethanol, 0.57 mM cysteine, 10 IU mL-1 pregnant mare serum gonadotropin, 10 IU mL-1 hCG, and 1 �M PGI2 analog for 22 h, and then further cultured in maturation medium without PGI2 analog and hormones for 22 h. After fertilization in Tris-buffered (mTBM) medium for 6 h, presumptive porcine zygotes were cultured in the NCSU-23 medium supplemented with 4% BSA for 6 days. All data were analyzed by using the Duncan test of ANOVA by the Statistical Analysis System (SAS Institute, Inc., Cary, NC, USA). First, we confirmed that PGI2 analog-treated (90.0 � 2.6%) oocytes showed a higher proportion of the metaphase II stage than non-treated (65.7 � 1.4%) ones (P &lt; 0.05). Thus, to confirm the activities of maturation promoting factor (MPF) and mitogen-activated protein kinase (MAPK), Western blots were performed in matured oocytes by using specific antibodies such as anti-cdc2 and anti-ERK1/2. The activities of MPF and MAPK were increased in porcine oocytes treated with PGI2 analog. In the PGI2 analog-treated group, polyspermic rate (17.9 � 13.3%) was reduced as compared with that of the non-treated group (35.8 � 9.4%). Furthermore, the rate (25.3%, 40/158) of blastocyst formation in the PGI2 analog-treated group was higher than in the non-treated group (19.7%, 27/137; P &lt; 0.05). Also, cell numbers of blastocysts were increased (29 � 2.5% vs. 39.6 � 1.4%) in the treated vs. the non-treated group. The numbers of fragmented DNA nuclei detected in the blastocyst stage by the TUNEL assay were decreased in the PGI2-treated group compared with the non-treated group (2.1% vs. 5.2%). In conclusion, direct roles of PGI2 during porcine oocyte maturation may involve reducing apoptosis and enhancing blastocyst quality.

scholarly journals Nucleus, Cytoskeleton, and Mitogen-Activated Protein Kinase p38 Dynamics during In Vitro Maturation of Porcine Oocytes

Animals ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 163
Author(s):  
Payungsuk Intawicha ◽  
Li-Kuang Tsai ◽  
Shih-Ying Yen ◽  
Neng-Wen Lo ◽  
Jyh-Cherng Ju
Keyword(s):  
Oocyte Maturation ◽  
Mammalian Cells ◽  
In Vitro Maturation ◽  
Germinal Vesicle ◽  
Mitogen Activated Protein Kinase ◽  
Meiotic Maturation ◽  
Germinal Vesicle Breakdown ◽  
Mitogen Activated Protein ◽  
Porcine Oocyte

The mitogen-activated kinase (MAPK) p38, a member of the MAPK subfamily, is conserved in all mammalian cells and plays important roles in response to various physiologic cues, including mitogens and heat shock. In the present study, MAPK p38 protein expression in porcine oocytes was analyzed during in vitro maturation (IVM) by Western blotting and immunocytochemistry. The levels of p-p38 or activated p38 and p38 expression were at the lowest in the germinal vesicle (GV) stage oocyte, gradually rising at the germinal vesicle breakdown (GVBD) and then reaching a plateau throughout the IVM culture (p < 0.05). Similarly, the expression level of total p38 was also lower in the GV oocyte than in the oocyte of other meiotic stages and uprising after GVBD and remained high until the metaphase III (MII) stage (p < 0.05). In the GV stage, phosphorylated p38 (p-p38) was initially detectable in the ooplasm and subsequently became clear around the nucleus and localized in the ooplasm at GVBD (18 h post-culture). During the metaphase I (MI) and metaphase II (MII) stages, p-p38 was evenly distributed throughout the ooplasm after IVM for 30 or 42 h. We found that the subcellular localization increased in p-p38 expression throughout oocyte maturation (p < 0.05) and that dynamic reorganization of the cytoskeleton, including microfilaments and microtubules, was progressively changed during the course of meiotic maturation which was likely to be associated with the activation or networking of p38 with other proteins in supporting oocyte development. In conclusion, the alteration of p38 activation is essential for the regulation of porcine oocyte maturation, accompanied by the progressive reorganization and redistribution of the cytoskeleton and MAPK p38, respectively, in the ooplasm.

scholarly journals Tributyltin Oxide Exposure During in vitro Maturation Disrupts Oocyte Maturation and Subsequent Embryonic Developmental Competence in Pigs

2021 ◽  
Vol 9 ◽  
Author(s):  
Yue Xiao ◽  
Bao Yuan ◽  
Weiyi Hu ◽  
Jiajia Qi ◽  
Hao Jiang ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Iron Homeostasis ◽  
Polar Body ◽  
Developmental Competence ◽  
Limited Information ◽  
Cellular Iron ◽  
Ros Accumulation ◽  
Porcine Oocyte ◽  
Cellular Iron Homeostasis

Tributyltin oxide (TBTO), an organotin compound, has been demonstrated to have toxic effects on several cell types. Previous research has shown that TBTO impairs mouse denuded oocyte maturation. However, limited information is available on the effects of TBTO exposure on livestock reproductive systems, especially on porcine oocytes in the presence of dense cumulus cells. In the present research, we evaluated the effects of TBTO exposure on porcine oocyte maturation and the possible underlying mechanisms. Porcine cumulus-oocyte complexes were cultured in maturation medium with or without TBTO for 42 h. We found that TBTO exposure during oocyte maturation prevented polar body extrusion, inhibited cumulus expansion and impaired subsequent blastocyst formation after parthenogenetic activation. Further analysis revealed that TBTO exposure not only induced intracellular reactive oxygen species (ROS) accumulation but also caused a loss of mitochondrial membrane potential and reduced intracellular ATP generation. In addition, TBTO exposure impaired porcine oocyte quality by disrupting cellular iron homeostasis. Taken together, these results demonstrate that TBTO exposure impairs the porcine oocyte maturation process by inducing intracellular ROS accumulation, causing mitochondrial dysfunction, and disrupting cellular iron homeostasis, thus decreasing the quality and impairing the subsequent embryonic developmental competence of porcine oocytes.

scholarly journals Curcumin supplementation in the maturation medium improves the maturation, fertilisation and developmental competence of porcine oocytes

2020 ◽  
Author(s):  
Zhao Namula ◽  
Yoko Sato ◽  
Manita Wittayarat ◽  
Quynh Anh Le ◽  
Nhien Thi Nguyen ◽  
...  
Keyword(s):  
Treated Group ◽  
Developmental Competence ◽  
Negative Effects ◽  
Beneficial Effects ◽  
Positive Effects ◽  
Fragmentation Index ◽  
Maturation Medium ◽  
Porcine Oocyte ◽  
Maturation Rate

AbstractThis study was conducted to determine the effects of supplementing the maturation medium with the antioxidant curcumin on the in vitro maturation (IVM), fertilisation and development of porcine oocytes. Curcumin supplementation was performed at concentrations of 0, 5, 10, 20, and 40 µM. At concentrations of 5–20 µM, curcumin had significant positive effects (P < 0.05) on maturation and fertilisation rates compared to the non-treated group. Of the groups cultured with 5–20 µM curcumin, the number of oocytes with DNA-fragmented nuclei after IVM was significantly lower than in groups matured without curcumin. Moreover, curcumin supplementation at 10 µM also gave a significantly higher rate of blastocyst formation compared with oocytes matured without curcumin. Increasing the curcumin concentration to 40 µM yielded negative effects on fertilisation and embryonic development compared with the groups treated with lower concentrations of curcumin. Supplementation with 10 µM curcumin had beneficial effects on the oocyte maturation rate and DNA fragmentation index compared to the non-treated group both in the presence and absence of hydrogen peroxide. These results indicate that curcumin supplementation at a suitable concentration (10 µM) is potentially useful for porcine oocyte culture systems, in terms of protecting oocytes from various forms of oxidative stress.

299 LEPTIN ACCELERATES PRONUCLEAR FORMATION FOLLOWING INTRACYTOPLASMIC SPERM INJECTION OF PORCINE OOCYTES: POSSIBLE ROLE OF MITOGEN-ACTIVATED PROTEIN KINASE INACTIVATION

2007 ◽  
Vol 19 (1) ◽  
pp. 265
Author(s):  
Y. X. Jin ◽  
X. S. Cui ◽  
W. K. Chang ◽  
T. Kim ◽  
N. H. Kim
Keyword(s):  
Protein Kinase ◽  
Oocyte Maturation ◽  
Intracytoplasmic Sperm Injection ◽  
Treated Group ◽  
Mitogen Activated Protein Kinase ◽  
Parthenogenetic Activation ◽  
Mitogen Activated Protein ◽  
Cytoplasmic Maturation ◽  
Pronuclear Formation

Although evidence suggests that leptin modulates oocyte maturation in vitro, few studies have examined the direct effect of leptin on cytoplasmic maturation and pronuclear formation following fertilization of porcine oocytes.The aim of this studywas to determine microtubule and microfilament assembly following oocyte maturation, and to assess blastocyst formation, mitogen-activated protein kinase (MAPK) activity, and pronuclear formation following parthenogenetic stimuli or intracytoplasmic sperm injection (ICSI) of leptin-treated oocytes.A symmetrical shape of the meiotic spindle was considered to be the normal microtubule assembly, and a clear, thick microfilament area containing a second metaphase chromatin was considered to be morphologically normal. Significant differences were determined using Tukey's multiple range test with P &lt; 0.05 being considered significant. Addition of 10 ng mL-1 of leptin to the in vitro maturation (IVM) medium significantly increased the percentage of zygotes that developed to the blastocyst stage (26.1 � 2.1%; P &lt; 0.05) compared with that of the control (0 ng mL-1: 16.3 � 2.8%) or 1 ng mL-1 (18.6 � 2.6%) leptin treatment groups following parthenogenetic activation. After IVM for 44 h, the percentage of oocytes with normal microtubules was higher in the leptin (10 ng mL-1) group than in the control group (89.1 � 4.3% vs. 78.5 � 4.0%, respectively; P &lt; 0.05). However, the percentage of normal microfilament assembly was similar in the leptin-treated and control groups (85.6 � 4.3% and 82.9 � 5.0%, respectively). In oocytes matured in vitro in the presence of 10 ng mL-1 of leptin and subsequently induced by parthenogenetic activation via chemical stimulation, there was a significant increase in the formation of 2 pronuclei (2PN: 52.1 � 5.2%; P &lt; 0.05) at 6 h, compared with the control non-leptin-treated oocytes (40.7 � 4.0%). Similarly, addition of 10 ng mL-1 of leptin to the IVM medium also increased 2PN formation at the same time point following ICSI (leptin treatment: 47.9 � 4.0; control: 36.2 � 3.8%; P &lt; 0.05). The addition of 10 ng mL-1 of leptin to the IVM medium decreased the level of phospho-ERK1/2 at 6 and 9 h and was lower in the leptin-treated group compared with the control non-leptin-treated group (P &lt; 0.01). These results suggest that exogenous leptin enhances cytoplasmic maturation and pronuclear formation following fertilization, possibly via the MAPK pathway.

Paradoxical effects of kisspeptin: it enhances oocyte in vitro maturation but has an adverse impact on hatched blastocysts during in vitro culture

2012 ◽  
Vol 24 (5) ◽  
pp. 656 ◽  
Author(s):  
Islam M. Saadeldin ◽  
Ok Jae Koo ◽  
Jung Taek Kang ◽  
Dae Kee Kwon ◽  
Sol Ji Park ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Formation Rate ◽  
Early Period ◽  
Threshold Effect ◽  
Developmental Competence ◽  
Control Group ◽  
Maturation Medium ◽  
Paradoxical Effects

Kisspeptin (Kp) is best known as a multifunctional peptide with roles in reproduction, the cardiovascular system and cancer. In the present study the expression of kisspeptin hierarchy elements (KISS1, GNRH1 and LHB) and their receptors (KISS1R, GNRHR and LHCGR, respectively) in porcine ovary and in cumulus–oocyte complexes (COCs) were investigated, as were its effects on the in vitro maturation (IVM) of oocytes and their subsequent ability to sustain preimplantation embryo competence after parthenogenetic electrical activation. Kp system elements were expressed and affected IVM of oocytes when maturation medium was supplemented with 10–6 M Kp. Oocyte maturation, maternal gene expression (MOS, GDF9 and BMP15), blastocyst formation rate, blastocyst hatching and blastocyst total cell count were all significantly increased when oocytes were matured in medium containing Kp compared with the control group (without Kp). A Kp antagonist (p234) at 4 × 10–6 M interfered with this hierarchy but did not influence the threshold effect of gonadotrophins on oocyte maturation. FSH was critical and permissive to Kp action on COCs by increasing the relative expression of KISS1R. In contrast, Kp significantly increased apoptosis, the expression of pro-apoptotic gene, BAK1, and suppressed trophoblast outgrowths from hatched blastocysts cultured on feeder cells. The present study provides the first functional evidence of the Kp hierarchy in porcine COCs and its role in enhancing oocyte maturation and subsequent developmental competence in an autocrine–paracrine manner. However, Kp supplementation may have a harmful impact on cultured hatched blastocysts reflecting systemic or local regulation during the critical early period of embryonic development.

scholarly journals Insulin acts via mitogen-activated protein kinase phosphorylation in rabbit blastocysts

Reproduction ◽  
2004 ◽  
Vol 128 (5) ◽  
pp. 517-526 ◽  
Author(s):  
Anne Navarrete Santos ◽  
Sarah Tonack ◽  
Michaela Kirstein ◽  
Marie Pantaleon ◽  
Peter Kaye ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Glucose Transport ◽  
Mapk Signaling ◽  
Mitogen Activated Protein Kinase ◽  
Preimplantation Embryos ◽  
Mrna Levels ◽  
Glut4 Translocation ◽  
Mitogen Activated Protein ◽  
Rabbit Embryos

The addition of insulin during in vitro culture has beneficial effects on rabbit preimplantation embryos leading to increased cell proliferation and reduced apoptosis. We have previously described the expression of the insulin receptor (IR) and the insulin-responsive glucose transporters (GLUT) 4 and 8 in rabbit preimplantation embryos. However, the effects of insulin on IR signaling and glucose metabolism have not been investigated in rabbit embryos. In the present study, the effects of 170 nM insulin on IR, GLUT4 and GLUT8 mRNA levels, Akt and Erk phosphorylation, GLUT4 translocation and methyl glucose transport were studied in cultured day 3 to day 6 rabbit embryos. Insulin stimulated phosphorylation of the mitogen-activated protein kinase (MAPK) Erk1/2 and levels of IR and GLUT4 mRNA, but not phosphorylation of the phosphatidylinositol 3-kinase-dependent protein kinase, Akt, GLUT8 mRNA levels, glucose uptake or GLUT4 translocation. Activation of the MAPK signaling pathway in the absence of GLUT4 translocation and of a glucose transport response suggest that in the rabbit preimplantation embryo insulin is acting as a growth factor rather than a component of glucose homeostatic control.

Sign in / Sign up

Export Citation Format

Share Document