Mammalian oocyte maturation

Thibault, C.; Szöllősi, Dániel; Gérard, Micheline

doi:10.1051/rnd:19870701

Cited by 207 publications

(103 citation statements)

References 49 publications

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“…Cortical granules are distributed throughout the entire inner surface of the sphere of the oocyte during final maturation [10,17]. Exocytosis of cortical granules is the most common mechanism for the prevention of polyspermy.…”

Section: Discussionmentioning

confidence: 99%

In Vitro Fertilization and Cortical Granule Distribution of Bovine Oocytes Having Heterogeneous Ooplasm with Dark Clusters.

Nagano

Takahashi

Katagiri

1999

The Journal of Veterinary Medical Science

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ABSTRACT. In vitro maturation, fertilization and subsequent development of oocytes with homogeneous (category 1), or heterogeneous ooplasm (category 2) were investigated. No significant differences were observed in the nuclear maturation and total fertilization rates between the two categories. However, category 2 oocytes showed a higher normal fertilization rate due to their lower incidence of polyspermy as compared to category 1 oocytes. Electron microscopic study revealed that all category 2 oocytes had cortical granules lined up next to the plasma membrane, and that some category 1 oocytes still had small clusters of cortical granules after maturation. Although the proportion of cleaved zygotes was higher in category 2, the percentages of cleaved zygotes that developed to the blastocyst stage did not differ between the two categories. These results demonstrate that oocytes with heterogeneous ooplasm have a higher capacity for normal fertilization due to the reduction in polyspermy. This can be attributed to the normal distribution of cortical granules in category 2 oocytes after maturation. -KEY WORDS: bovine, cortical granule, fertilization, morphology.J. Vet. Med. Sci. 61(5): 531-535, 1999 Sigma Chemical Co., St. Louis, U.S.A.), 0.2 mM sodium pyruvate (Sigma) and 50 µg/ml gentamicin sulfate (Sigma). During the washing procedure, oocytes were examined, selected and classified under a stereomicroscope by the following criteria. Category 1 represented oocytes with homogeneous ooplasm and multilayered cumulus investment. Category 2 represented oocytes having heterogeneous ooplasm with dark clusters, and multilayered cumulus investment.In vitro maturation, fertilization and culture: In vitro maturation, fertilization and culture were carried out using the procedure described previously [15]. Briefly, the COCs were washed once in the maturation medium; 25 mM Hepesbuffered TCM 199 supplemented with 10% fetal calf serum (Gibco Laboratories, Grand Island, NY, U.S.A.), 0.02 units/ ml follicle-stimulating hormone (Sigma), 1 µg/ml estradiol-17β (Sigma) and 0.2 mM sodium pyruvate and 50 µg/ml gentamicin sulfate. The oocytes in each category were cultured separately for 22 hr in a 50 µl droplet of maturation medium (10 or 11 oocytes per droplet) covered with paraffin oil at 39°C in a humidified atmosphere of 5% CO 2 in air. Frozen bovine semen collected from a Holstein bull was used for in vitro fertilization. After thawing the semen in a 37°C water bath for 30 sec, the motile sperm were separated using a discontinuous gradient with 45 and 90% Percoll (Pharmacia BioProcess, Uppsala, Sweden) by centrifugation at 700 × g for 20 min. The sperm were subsequently washed with a modified Brackett and Oliphant (mBO) medium [3] by centrifugation at 500 × g for 5 min. The mature oocytes were co-incubated with the sperm at the concentration of 5 × 10 6 sperm/ml in a 100 µl droplet of mBO medium containing 3 mg/ml fatty-acid-free BSA (Sigma) and 2.5 mM theophylline (Sigma) covered with paraffin oil for 20 hr at 39°C in a humidif...

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Section: Discussionmentioning

confidence: 99%

In Vitro Fertilization and Cortical Granule Distribution of Bovine Oocytes Having Heterogeneous Ooplasm with Dark Clusters.

Nagano

Takahashi

Katagiri

1999

The Journal of Veterinary Medical Science

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“…Among the steps in IVP, in vitro maturation (IVM) is highlighted (Brackett & Zuelke, 1993;Eppig, 1996;Sirard & Blondin, 1996) because in this phase the oocyte accumulates the mRNA and proteins that are essential to progression to the embryonic genome activation stage and to the acquisition of developmental competence (Thibault et al, 1987;Sirard et al, 1989).…”

Section: Introductionmentioning

confidence: 99%

The effect of interaction between macromolecule supplement and oxygen tension on bovine oocytes and embryos culturedin vitro

Mingoti

Castro

Méo

et al. 2009

Zygote

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SummaryAiming to improve in vitro production of bovine embryos and to obtain supplements to replace serum for in vitro maturation (IVM), this study evaluated the effects of macromolecular supplementation of IMV medium (bovine serum albumin -BSA, polyvinyl alcohol -PVA, polyvinyl pyrrolidone -PVP, Ficoll, KnockoutSR, or fetal calf serum -FCS) and oxygen tension [5% CO 2 in air (20% O 2 ) or 5% CO 2 , 5% O 2 and 90% N 2 (5% O 2 )] on oocyte maturation and embryo development. Nuclear progression to germinal vesicle breakdown, metaphase I and metaphase II stages were evaluated and overall results revealed that undefined (FCS) and semi-defined (BSA) media gave better results at 20% O 2 and defined media (PVA, PVP and Ficoll) at 5% O 2 . Independent of macromolecule supplement, IVM at 20% O 2 was considered optimal for nuclear maturation. To evaluate embryo development, oocytes matured in the previously described conditions were fertilized and cultured at the same oxygen tension used for IVM and assessed for cleavage (43.0 to 74.8%) and development to morulae (16.4 to 33.8%), blastocyst (7.7 to 52.9%) and hatched blastocyst (9.6 to 48.1%). Apart from oxygen tension, all treatments, except Knockout (22.7%), gave similar results for blastocyst development (26.5 to 38.7%). Independently of macromolecule supplement, higher development rates were obtained in an oxygen tension of 20% O 2 (67.4% cleavage, 29.2% morulae, 40.8% blastocyst and 34.0% hatched blastocyst) when compared with 5% O 2 (52.5, 21.8, 18.2 and 15.6%, respectively). This study indicates that BSA, PVA, PVP and Ficoll can replace serum during IVM and that the optimal atmospheric condition for in vitro production of bovine embryos is 5% CO 2 and 20% O 2 .

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“…A further factor that may influence the success of enucleation in mice and ungulates is the difference in orientation of the first metaphase plate between these species. In mouse oocytes the spindle is organized in a parallel orientation to the membrane while in sheep, pig and cattle a perpendicular orientation is normal [24]. After ETO treatment, however, no spindle is detected and only a single cluster of chromatin can be seen [10].…”

mentioning

confidence: 99%

Chemically enucleated mouse oocytes: ultrastructure and kinetics of histone H1 kinase activity

Kárníková

Horská²,

Tománek³

et al. 1998

Reprod. Nutr. Dev.

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Mammalian oocyte maturation

Abstract: Eppig, 1977Eppig, , 1979Bachvarova et al., 1980

Cited by 207 publications

References 49 publications

In Vitro Fertilization and Cortical Granule Distribution of Bovine Oocytes Having Heterogeneous Ooplasm with Dark Clusters.

In Vitro Fertilization and Cortical Granule Distribution of Bovine Oocytes Having Heterogeneous Ooplasm with Dark Clusters.

The effect of interaction between macromolecule supplement and oxygen tension on bovine oocytes and embryos culturedin vitro

Chemically enucleated mouse oocytes: ultrastructure and kinetics of histone H1 kinase activity

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