Artificial activation of bovine oocytes matured in vitro by electric shock or exposure to ionophore A23187

Aoyagi, Yoshito; Kameyama, Kimiko; Takeda, T.

doi:10.1016/0093-691x(92)90257-r

Theriogenology

1992

DOI: 10.1016/0093-691x(92)90257-r

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Artificial activation of bovine oocytes matured in vitro by electric shock or exposure to ionophore A23187

Yoshito Aoyagi¹,

Kimiko Kameyama²,

T. Takeda³

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Cited by 7 publications

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“…Many of these treatments, such as strontium (Meo et al 2004), ionomycin (Hill et al 2000), calcium ionophore A23187 (Chung et al 2001, Sedmikova et al 2003, electrical pulses (Aoyagi et al 1992, Yang et al 1992, Prochazka et al 1993, Kure-bayashi et al 1996, Escriba and Garcia-Ximenez 1999 and ethanol (Nagai 1987, Minamihashi et al 1993, are able to induce oocytes activation by promoting an increase in intracellular free calcium concentration. Intracellular calcium transients have been associated with the early events of oocyte activation (White and Yue 1996) and seem to be required for CSF reduction and MPF inactivation by cyclin degradation.…”

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Artificial Meiotic Arrest and Parthenogenetic Activation of Buffalo Oocytes Using Cycloheximide

2007

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Summary Maintaining oocytes at germinal vesicle (GV) stage without damaging their quality would allow synchronization of maturation and homogenization of the oocyte population. Activation of the oocyte is very important for a number of oocyte or embryo related technologies including intracytoplasmic sperm injection and cloning by nuclear transfer. This work will focus on induction of meiotic inhibition and oocyte activation by cycloheximide (CHX), in buffalo. For this purpose 2 experiments were conducted. In experiment I, buffalo cumulus-oocyte complexes (COCs) were cultured in 100 ml droplets of tissue culture medium 199 enriched with 10% v/v fetal calf serum, FSH and LH (0, 05 iu/ml, PERGOVET 75, Serono, Rome, Italy) and 50 mg/ml gentamycin for 24-26 h at 38.5°C in 5% CO 2 and humidified air for 24 h in the presence of 2 mg/ml CHX. COCs were significantly blocked at the GV stage (20.85% vs. 82.72% in control). Reversibility of the CHX effect was assessed by culturing COCs an additional 24 h in CHX-free culture medium. About 65.0% of treated oocytes (control 80.1%) resumed meiosis and progressed to the MII stage. In experiment II, buffalo oocytes were activated by 10 mM calcium ionophore A23187 for 5 min followed by 10 mg/ml CHX for 3 h. Control oocytes were exposed to conventional in vitro fertilization (IVF) in BO medium. After 48 h of culture, significantly more oocytes (pϽ0.05) were cleaved in the IVF group than in treated group and spontaneous activated one. In conclusion, CHX can be used to block spontaneous resumption of meiosis in buffalo oocytes and its effect is reversible. Also, CHX in combination with calcium ionophore have the ability to induce parthenogenetic activation in buffalo oocytes.

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Artificial Meiotic Arrest and Parthenogenetic Activation of Buffalo Oocytes Using Cycloheximide

2007

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Development of an efficient method to produce uniformly haploid parthenogenones

1999

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Electrically induced calcium elevation, activation, and parthenogenetic development of bovine oocytes

Collas¹,

Fissore

Robl

et al. 1993

Molecular Reproduction Devel

115

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The influence of electrical stimulation on the level of intracellular Ca2+ in bovine oocytes, as well as activation and extent of parthenogenetic development, was investigated. Mature oocytes were electrically stimulated at 29 hr of maturation, and intracellular Ca2+ concentration was determined with the Ca2+ indicator fura-2 dextran (fura-2 D). The Ca2+ response of oocytes to a given electrical pulse was variable. Oocytes responded with either no Ca2+ rise from baseline (approximately 12 nM), a short-duration Ca2+ rise (from 12 nM to 300 nM) that returned to baseline within 2 min of the pulse, or a long-duration Ca2+ rise (from 12 nM to 1,000-2,000 nM) that never returned to baseline during the 8 min period over which the oocytes were monitored. In these oocytes, Ca2+ level returned to baseline when oocytes were removed from 0.30 M mannitol and placed in an ionic medium. Increasing field strength or pulse duration tended to increase the proportion of oocytes displaying a Ca2+ rise, and at 1.0 kVcm-1 for 40 microseconds, all oocytes displayed a long-duration Ca2+ elevation. Direct transfer of oocytes from culture medium to mannitol also triggered a Ca2+ rise. Multiple stimulations, either electrical or by transferring to mannitol, produced multiple Ca2+ rises. This mannitol-induced Ca2+ rise could be inhibited by first washing the oocytes in medium containing equal parts of 0.30 M mannitol and phosphate buffered saline (PBS). The level of Ca2+ stimulation affected activation and development of oocytes. Insufficient, or, conversely, excessive Ca2+ stimulation impaired development. Optimum development was obtained with 1) three pulses of 0.2 kVcm-1 for 20 microseconds, each pulse 22 min apart, after direct transfer of oocytes from culture medium to mannitol (22% blastocysts) or 2) three pulses of 1.0 kVcm-1 for 20 microseconds after transfer of oocytes from culture medium to medium containing equal parts mannitol and PBS, then to mannitol (24% blastocysts). This procedure avoided induction of a Ca2+ rise prior to the pulse. The results indicate that the level of Ca2+ stimulation can be regulated by incubation conditions prior to the pulse and, to some extent, by field strength and pulse duration. The level of electrical stimulation influenced oocyte Ca2+ response, activation, and parthenogenetic development.

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Artificial activation of bovine oocytes matured in vitro by electric shock or exposure to ionophore A23187

Cited by 7 publications

References 0 publications

Artificial Meiotic Arrest and Parthenogenetic Activation of Buffalo Oocytes Using Cycloheximide

Artificial Meiotic Arrest and Parthenogenetic Activation of Buffalo Oocytes Using Cycloheximide

Development of an efficient method to produce uniformly haploid parthenogenones

Electrically induced calcium elevation, activation, and parthenogenetic development of bovine oocytes

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