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Cytoplasmic Maturation (cytoplasmic + maturation)
Selected AbstractsElevated Histone H1 (MPF) and Mitogen-activated Protein Kinase Activities in Pig Oocytes Following In Vitro Maturation do not Indicate Cytoplasmic MaturationREPRODUCTION IN DOMESTIC ANIMALS, Issue 2 2009MA Setiadi Contents Effects of different media (TCM 199 + BSA, TCM 199 + FCS, TCM 199 + NBCS, Whitten's medium + BSA) supplemented with estradiol-17, and two isolated and everted follicle shells on MPF and MAP kinase activities and the sensitivity to parthenogenetic activation of pig oocytes were examined at the end of culture (48 h). Elevated (P < 0.05) activities of MAP kinase were recorded in metaphase II oocytes following culture in Whitten's medium, whereas MPF levels were lowest (P < 0.05) in MII oocytes matured in TCM 199 supplemented with BSA. Oocytes matured in TCM 199 based media showed higher (P < 0.05) activation rates when compared to oocytes incubated in Whitten's medium. Whitten's medium supplemented with different protein sources (amino acids, FCS, BSA) was used to study the effects of different exposure periods to eCG/hCG stimulation on MPF and MAP kinase activities and in vivo fertilisability following culture for 48 h. MPF and MAP kinase activities were significantly increased by eCG/hCG stimulation of COCs during maturation. Further, the continuous presence of eCG/hCG during culture (48 h) significantly increased the levels of both kinases in comparison to stimulation by gonadotrophins alone during the first 24 h of incubation. In vivo fertilisation of oocytes matured in Whitten's medium supplemented with eCG/hCG for 24 or 48 h led to a significant retardation of early embryonic development compared to ovulated oocytes. In conclusion, media composition and gonadotrophin stimulation affect MPF/MAP kinase activities and the susceptibility to parthenogenetic activation of IVM oocytes. However, elevated kinase levels in pig oocytes following culture do not indicate complete cytoplasmic maturation. [source] Ion current activity and molecules modulating maturation and growth stages of ascidian (Ciona intestinalis) oocytesMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 11 2009Francesco Silvestre Electrophysiological techniques were used to study ion currents in the ascidian Ciona intestinalis oocyte plasma membranes during different stages of growth and meiosis. Three stages (A, B, C) of immature oocytes were discriminated in the ovary, with the germinal vesicle (GV) showing specific different features of growth and maturation. Stage A (pre-vitellogenic) oocytes exhibited the highest L-type Ca2+current activity, and were incompetent for meiosis resumption. Stage B (vitellogenic) oocytes showed Na+ currents that remained high during the maturation, up to the post-vitellogenic stage C oocytes. The latter had acquired meiotic competence, undergoing spontaneous maturation and interacting with the spermatozoon. However, fertilized oocytes did not produce normal larvae, suggesting that cytoplasmic maturation plays a specific role in embryo development. Spontaneous maturation was inhibited at low pH whereas trypsin was able to trigger germinal vesicle breakdown (GVBD) regardless of pH; in addition spontaneous maturation was not affected by removal of follicle cells or by inhibiting junctional communication between oocyte and follicle cells. Taken together these results imply: (i) Ca2+ and Na+ currents are involved in meiotic progression, growth, and acquisition of meiotic competence; (ii) trypsin-like molecules may have a role as candidates for providing the physiological stimulus to resume meiosis. Finally, we provide evidence that follicle cells in Ciona are not involved in triggering GVBD as it occurs in other ascidians. Mol. Reprod. Dev. 76: 1084,1093, 2009. © 2009 Wiley-Liss, Inc. [source] Effects of thiol compounds on in vitro maturation of canine oocytes collected from different reproductive stagesMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 9 2007Mohammad Shamim Hossein Abstract Various thiol compounds are known to improve cytoplasmic and/or nuclear maturation of oocytes in vitro. The present study examined the effects of two thiol compounds, cysteine (0.1, 0.5, and 1.0 mM) and cysteamine (50, 100, and 200 µM), on cytoplasmic and nuclear maturation of canine oocytes. Oocytes collected from different reproductive stages were cultured in TCM-199 supplemented with 10% fetal bovine serum, 2.2 mg/ml sodium carbonate, 2.0 µg/ml estrogen, 0.5 µg/ml FSH, 0.03 IU/ml hCG, and 1% penicillin,streptomycin solution for 72 h. Data were analyzed by two-way ANOVA after arcscine transformation and protected by Bonferroni post hoc test. The effects of cysteine and cysteamine on canine IVM were varied depending on the reproductive stage of oocyte donor bitches. In the follicular stage, significantly more oocytes reached the metaphase II (M II) stage when cultured with 0.5 or 1.0 mM cysteine (16.7% and 16.9%, respectively) compared to the control (6.2%). In the follicular stage, cysteamine increased oocyte maturation rate upto the M II stage (15.1% to 17.0%) compared to the control (4.4%). Both the 0.5 mM cysteine and 100 µM cysteamine, alone or together, increased the intracellular GSH level of canine oocytes compared to the control. Irrespective of reproductive stage, no further beneficial effects on nuclear or cytoplasmic maturation were observed when 0.5 mM cysteine and 100 µM cysteamine were supplemented together. In conclusion, addition of 0.5 mM cysteine and 100 µM cysteamine to the maturation medium improved IVM of canine oocytes. Mol. Reprod. Dev. 74: 1213,1220, 2007. © 2007 Wiley-Liss, Inc. [source] Elevated Histone H1 (MPF) and Mitogen-activated Protein Kinase Activities in Pig Oocytes Following In Vitro Maturation do not Indicate Cytoplasmic MaturationREPRODUCTION IN DOMESTIC ANIMALS, Issue 2 2009MA Setiadi Contents Effects of different media (TCM 199 + BSA, TCM 199 + FCS, TCM 199 + NBCS, Whitten's medium + BSA) supplemented with estradiol-17, and two isolated and everted follicle shells on MPF and MAP kinase activities and the sensitivity to parthenogenetic activation of pig oocytes were examined at the end of culture (48 h). Elevated (P < 0.05) activities of MAP kinase were recorded in metaphase II oocytes following culture in Whitten's medium, whereas MPF levels were lowest (P < 0.05) in MII oocytes matured in TCM 199 supplemented with BSA. Oocytes matured in TCM 199 based media showed higher (P < 0.05) activation rates when compared to oocytes incubated in Whitten's medium. Whitten's medium supplemented with different protein sources (amino acids, FCS, BSA) was used to study the effects of different exposure periods to eCG/hCG stimulation on MPF and MAP kinase activities and in vivo fertilisability following culture for 48 h. MPF and MAP kinase activities were significantly increased by eCG/hCG stimulation of COCs during maturation. Further, the continuous presence of eCG/hCG during culture (48 h) significantly increased the levels of both kinases in comparison to stimulation by gonadotrophins alone during the first 24 h of incubation. In vivo fertilisation of oocytes matured in Whitten's medium supplemented with eCG/hCG for 24 or 48 h led to a significant retardation of early embryonic development compared to ovulated oocytes. In conclusion, media composition and gonadotrophin stimulation affect MPF/MAP kinase activities and the susceptibility to parthenogenetic activation of IVM oocytes. However, elevated kinase levels in pig oocytes following culture do not indicate complete cytoplasmic maturation. [source] Improving in vitro Maturation of Oocytes in the Human Taking Lessons from Experiences in Animal SpeciesREPRODUCTION IN DOMESTIC ANIMALS, Issue 1 2001J Smitz One to three per cent of infertile women develop severe ovarian hyperstimulation syndrome after superovulation for assisted reproduction treatment (ART). This severe complication can be avoided when oocytes are obtained at an immature stage (germinal vesicle stage) out of small or medium-sized follicles. This hypothesis has been tested in several infertile women, but clinical pregnancies are disappointlingly low. This new approach in ART is still at an experimental phase and this treatment has still to be improved before routine clinical application. Experimental work in animals and humans suggest a beneficial effect in providing a short preliminary pretreatment with follicle-stimulating hormone to select for a developing cohort of follicles. The aspiration of oocyte cumulus complexes is carried out with a short needle applying reduced aspiration pressure. A crucial point is to provide the appropriate culture environment for the immature oocytes. An optimal cumulus-enclosed human oocyte culture system needs to be defined. The composition of the culture medium could be suggested by in vitro work carried out in animal models. As developmental competence is established during the latest phases of oocyte growth and is dependent on the storage of RNA, a prolonged in vitro maturation period (before inducing nuclear maturation) could provide the necessary transcriptional and translational changes. The conditions to achieve this improved cytoplasmic maturation by prolonging the in vitro culture remain to be defined. More objective noninvasive parameters for oocyte maturity are also needed to pursue research in this field. [source] Early metaphase II oocytes treated with dibutyryl cyclic adenosine monophosphate provide suitable recipient cytoplasm for the production of miniature pig somatic cell nuclear transfer embryosANIMAL SCIENCE JOURNAL, Issue 1 2010Satoshi SUGIMURA ABSTRACT We investigated the effects of in vitro maturation duration and treatment with dibutyryl cyclic adenosine monophosphate (dbcAMP) on the blind enucleation efficiency and developmental competence of miniature pig somatic cell nuclear transfer (SCNT) embryos. Oocytes were cultured for 22 h in NCSU-23 medium with or without 1 mM dbcAMP and then additionally cultured in dbcAMP-free NCSU-23 for 14, 18, or 22 h. Regardless of dbcAMP treatment, the rate of nuclear maturation reached a plateau at 36 and 40 h. However, mitochondrial distribution, a marker for cytoplasmic maturation, differed between the dbcAMP-untreated oocytes at 36 h and dbcAMP-treated oocytes at 40 h. The metaphase II chromosomes were adjacent to the first polar body in 68.8% and 63.5% of the dbcAMP-untreated oocytes at 36 h and dbcAMP-treated oocytes at 40 h, respectively. Furthermore, the blind enucleation efficiency by removing a small volume of cytoplasm was significantly higher in the dbcAMP-untreated oocytes at 36 h (82.9%) and dbcAMP-treated oocytes at 40 h (89.9%) than other groups. The rate of blastocyst formation was highest in the dbcAMP-treated oocytes at 40 h. Hence, this study demonstrated that dbcAMP-treated early metaphase II oocytes are suitable for the production of miniature pig SCNT embryos. [source] | ||||||||||