Quercetin is a plant-derived flavonoid found in fruits or vegetables that has antioxidant properties and acts as a free radical scavenger. We investigated the effects of quercetin on porcine oocyte nuclear maturation and embryonic development after parthenogenetic activation. We then evaluated the antioxidant activities of quercetin by measuring reactive oxygen species (ROS) levels in matured oocytes. Immature oocytes were untreated or treated with 1, 10, and 50 microg/mL quercetin during in vitro maturation (IVM). Quercetin treatment did not improve oocyte nuclear maturation, but significantly higher blastocyst rates (p < 0.05) of parthenogenetically activated oocytes were achieved when the IVM medium was supplemented with an adequate concentration of quercetin (1 microg/mL). However, cleavage rates and blastocyst cell numbers were not affected. Oocytes treated with 1 or 10 microg/mL quercetin had significantly lower (p < 0.05) levels of ROS than the control and group treated with the highest concentration of quercetin (50 microg/mL). Moreover, this highest concentration was detrimental to oocyte nuclear maturation and blastocyst formation. Based on our findings, we concluded that exogenous quercetin reduces ROS levels during oocyte maturation and is beneficial for subsequent embryo development.
Animals ; Antioxidants/administration & dosage/*pharmacology ; Dose-Response Relationship, Drug ; In Vitro Oocyte Maturation Techniques/*veterinary ; Oocytes/cytology/*drug effects/physiology ; Quercetin/administration & dosage/*pharmacology ; Reactive Oxygen Species/*metabolism ; *Swine

Quercetin is a plant-derived flavonoid found in fruits or vegetables that has antioxidant properties and acts as a free radical scavenger. We investigated the effects of quercetin on porcine oocyte nuclear maturation and embryonic development after parthenogenetic activation. We then evaluated the antioxidant activities of quercetin by measuring reactive oxygen species (ROS) levels in matured oocytes. Immature oocytes were untreated or treated with 1, 10, and 50 microg/mL quercetin during in vitro maturation (IVM). Quercetin treatment did not improve oocyte nuclear maturation, but significantly higher blastocyst rates (p < 0.05) of parthenogenetically activated oocytes were achieved when the IVM medium was supplemented with an adequate concentration of quercetin (1 microg/mL). However, cleavage rates and blastocyst cell numbers were not affected. Oocytes treated with 1 or 10 microg/mL quercetin had significantly lower (p < 0.05) levels of ROS than the control and group treated with the highest concentration of quercetin (50 microg/mL). Moreover, this highest concentration was detrimental to oocyte nuclear maturation and blastocyst formation. Based on our findings, we concluded that exogenous quercetin reduces ROS levels during oocyte maturation and is beneficial for subsequent embryo development.
Animals ; Antioxidants/administration & dosage/*pharmacology ; Dose-Response Relationship, Drug ; In Vitro Oocyte Maturation Techniques/*veterinary ; Oocytes/cytology/*drug effects/physiology ; Quercetin/administration & dosage/*pharmacology ; Reactive Oxygen Species/*metabolism ; *Swine

OBJECTIVETo study the effects of the co-administration of growth hormone (GH) and aspirin to women with suboptimal response to GnRHa/FSH hyperstimulation protocol during in vitro fertilization and embryo transfer (IVF-ET) cycles.

METHODSForty cases of poor ovarian response in previous IVF-ET cycles were randomly divided into 2 groups: the studied group of GH and aspirin (n = 20), and the control group without GH or aspirin (n = 20).

RESULTSThe co-administration of GH and aspirin significantly increased the rates of retrieved oocytes (P < 0.01), promoted the maturation of oocytes (P < 0.01) and improve the fertilization rates (P < 0.05). However, there were no statistically differences between the two groups in the number of replaced embryos (P > 0.05) and the pregnancy rate (P > 0.05).

CONCLUSIONThe co-administration of GH and aspirin to poor ovarian responders is effective to increase the rates of retrieved oocytes, promote the maturation of oocytes and improve the fertilization rate in IVF-ET.

Aspirin ; administration & dosage ; Embryo Transfer ; Female ; Fertilization in Vitro ; Growth Hormone ; administration & dosage ; Humans ; Infertility, Female ; drug therapy ; physiopathology ; Oocytes ; cytology ; drug effects ; physiology ; Ovulation Induction ; Pregnancy ; Treatment Outcome

OBJECTIVETo investigate the potential of siRNAs targeting sphingomyelin phosphodiesterase 1 (SMPD1) in protecting the oocytes from apoptosis, and explore new approaches to female fertility preservation.

METHODSChemically synthesized siRNA targeting SMPD1 were introduced into mouse oocytes retrieved by hyperstimulation, and the cell apoptosis was analyzed by comic assay 48 and 72 h later.

RESULTSIn the oocytes without any siRNA injection, oocyte DNA damage occurred after 24 h, and large amount of DNA fragments migrated from the cells 48 h later. In oocytes injected with siRNA003, DNA migration decreased significantly as compared with the control and the other two groups injected with siRNA001 and siRNA002 (P<0.01).

CONCLUSIONsiRNA targeting SMPD1 may protect the oocytes from apoptosis, and has the potential for use in future female fertility preservation.

Animals ; Apoptosis ; drug effects ; genetics ; Comet Assay ; Female ; Mice ; Oocytes ; cytology ; RNA Interference ; RNA, Small Interfering ; genetics ; Sphingomyelin Phosphodiesterase ; genetics ; physiology ; Transfection

The purpose of this study was to investigate whether sperm selection by hyaluronic acid (HA) binding could improve fertilization rate and embryo quality in intracytoplasmic sperm injection (ICSI) cycles. Two hundred nineteen oocytes obtained from eighteen women were injected with either HA-bound (n = 107) or conventionally selected spermatozoa (n = 112) in a randomized way. All of the participants were infertile couples who had normal sperm parameters but low fertilization rate in previous in vitro fertilization (IVF) cycle (n = 5) or experienced multiple IVF failures (n = 13). Lower fertilization (75.7% vs 83.0%) and cleavage rate on day 2 (72.9% vs 83.0%) was observed in oocytes injected with HA-bound spermatozoa than the conventional group, but the difference was not significant. Significantly lower cleavage rate was observed on day 3 in HA group (56.0% vs 69.6%, P = 0.038). Blastocyst formation rate and the number of transferred embryos were similar in both groups. In multiple IVF failure patients, significantly reduced fertilization rate (71.8% vs 85.3%, P = 0.046) and cleavage rate on day 2 (70.4% vs 85.3%, P = 0.029) and day 3 (53.5% vs 77.3%, P = 0.002) were noticed in HA group. Five women achieved pregnancy continuing more than 12 weeks after transfer (27.8%). Success of ICSI was not related with the number of embryos fertilized by HA-bound spermatozoa. Application of ICSI by sperm selection using HA binding is not helpful in couples with repeated poor fertilization or implantation despite normal sperm parameters.
Adult ; Blastocyst/cytology ; Embryo Transfer ; Female ; *Fertilization in Vitro ; Humans ; Hyaluronic Acid/*pharmacology ; Infertility, Male/therapy ; Male ; Oocytes/cytology/physiology ; Pregnancy ; Pregnancy Rate ; Prospective Studies ; *Sperm Injections, Intracytoplasmic ; Spermatozoa/*drug effects/physiology

AIMThe mechanisms of cytokines in regulating oocyte maturation is still little known. The present study attempt to investigate whether the protooncogene of c-erbB2, c-myb are involved in introducing of cytokines to regulate oocyte maturation.

METHODSThis research used mouse GV stage oocyte culture model in vitro and RT-PCR, Western blotting method to explore the effect of EGF, TNFalpha, ET-1 and NO on oocyte maturation; to analyze the c-erbB2 mRNA and c-myb mRNA expression and the phosphorylation of MAPK and cyclinB1 expression in oocytes affected by above cytokines.

RESULTSEGF(10 microg/L) stimulated meiosis of oocytes significantly, the level of c-erbB2 mRNA, c-myb mRNA were increased, and promoted the phosphorylation of MAPK and cyclinB1 expression; TNFalpha (1 microg/L) and ET-1 ((10(-1) mol/L) had the results to EGF. Low dose of SNP (10(-5)mol/L) had no effect on oocyte maturation, but could significantly reverse the suppression of dbcAMP on oocyte maturation.

CONCLUSIONc-erbB2 and c-myb were involved in introducing of cytokines to regulate oocyte maturation, might be the middle link in connection of the cytokines with MAPK and MPF in regulation oocyte maturation.

Animals ; Cells, Cultured ; Cytokines ; physiology ; Epidermal Growth Factor ; physiology ; Female ; Intercellular Signaling Peptides and Proteins ; physiology ; Maturation-Promoting Factor ; genetics ; metabolism ; Mice ; Mitogen-Activated Protein Kinases ; metabolism ; Oocytes ; cytology ; growth & development ; physiology ; Oogenesis ; drug effects ; physiology ; Proto-Oncogene Proteins c-myb ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Receptor, ErbB-2 ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; physiology

Effect of sodium nitroprusside (SNP), a nitric oxide (NO) donor, on in vitro survival, growth, steroidogenesis, and apoptosis of buffalo preantral follicles (PFs) was investigated. PFs (200~250 microm) were isolated by micro-dissection and cultured in 0 (control), 10(-3), 10(-5), 10(-7), and 10(-9) M SNP. To examine the reversible effect of SNP, PFs were cultured with 10(-5) M SNP + 1 mM Nomega-nitro-L-arginine methyl ester (L-NAME) or 1.0 microg hemoglobin (Hb). The results showed that greater concentrations of SNP (10(-3), 10(-5), 10(-7) M) inhibited (p < 0.05) FSH-induced survival, growth, antrum formation, estradiol production, and oocyte apoptosis in a dose-dependent manner. However, a lower dose of SNP (10(-9) M) significantly stimulated (p < 0.05) the survival, growth, antrum formation, follicular oocyte maturation, and stimulated progesterone secretion compared to the control. A combination of SNP + L-NAME promoted the inhibitor effect of SNP while a SNP + Hb combination reversed this effect. Nitrate and nitrite concentrations in the culture medium increased (p < 0.05) in a dose-dependent manner according to SNP concentration in the culture medium. At higher concentrations, SNP had a cytotoxic effect leading to follicular oocyte apoptosis whereas lower concentrations have stimulatory effects. In conclusion, NO exerts a dual effect on its development of buffalo PFs depending on the concentration in the culture medium.
Animals ; *Apoptosis ; Buffaloes/*physiology ; Estradiol/biosynthesis ; Female ; Follicle Stimulating Hormone/metabolism ; NG-Nitroarginine Methyl Ester/pharmacology ; Nitrates/pharmacology ; Nitric Oxide/*metabolism ; Nitric Oxide Donors/pharmacology ; Nitrites/pharmacology ; Nitroprusside/pharmacology ; Oocytes/cytology/drug effects/growth & development/metabolism ; Ovarian Follicle/*cytology/drug effects/growth & development/*metabolism ; Progesterone/biosynthesis

For parthenogenetic activation as a model system of nuclear transfer, microinjection and electroporation as activation treatments in bovine metaphase II oocytes were administered to each of three groups as follows: control group (treatments with Ca2+, Mg2+ -free PBS+100 micro M EGTA), IP3 group (control+25 micro M IP3) and IP3+ ryanodine group (control+25 micro M IP3+10 mM ryanodine). In experiments using microinjection, no significant differences were observed between any of the developmental stages of the electroporation experiment. For electroporation, cleavage rates were significantly higher in the IP3+ryanodine group than in the IP3 or control group (85.6% vs 73.7% or 67.6%, respectively). In the subsequent stages of embryonic development, such as morula and blastocyst formation, the IP3 and ryanodine group exhibited significantly higher rates of morula fomation than the IP3 or control groups (40.6% vs 24.2% or 16.7%, respectively). Similarly, the rate of blastocyst formation in the IP3+ryanodine group was significantly higher than the control group (16.3% vs 6.9%) but did not differ significantly from the IP3 group (16.3% vs 9.5%). In nuclear transfer, activation was performed at 30 hpm by microinjection and elecroporation with 25 micro M IP3+ 10 mM ryanodine followed by 6-DMAP treatment. No significant differences were observed at any stage of embryonic development and none of the embryos activated by electroporation reached either the morula or blastocyst stage. However, 3.8% and 1.9% of embryos activated by microinjection sucessfully developed to the morula and blastocyst stages, respectively. In conclusion, activation treatments using IP3 and ryanodine are able to support the development of bovine parthenogenetic and reconstructed embryos.
Adenine/administration & dosage/*analogs & derivatives/pharmacology ; Animals ; Cattle/*embryology/physiology ; Cell Fusion ; Electroporation/veterinary ; Embryonic and Fetal Development/*drug effects ; Enzyme Inhibitors/administration & dosage/pharmacology ; Female ; Inositol 1,4,5-Trisphosphate/administration & dosage/*pharmacology ; Microinjections/veterinary ; Nuclear Transfer Techniques ; Oocytes/drug effects/growth & development ; Parthenogenesis/*drug effects ; Protein Kinase Inhibitors ; Ryanodine/administration & dosage/*pharmacology ; Skin/cytology