Inhibition of Protein Kinases after an Induced Calcium Transient Causes Transition of Bovine Oocytes to Embryonic Cycles without Meiotic Completion

Susko-Parrish, J.L.; Leibfried-Rutledge, M.L.; Northey, D.L.; Schutzkus, V.; First, N. L.

doi:10.1006/dbio.1994.1351

Cited by 258 publications

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“…Slimane et al [26] reported that 34.2% of two-cell IVF embryos had abnormalities such as haploidy, mixoploidy, hyperploidy and hypoploidy. Previous studies have examined pronuclear formation status and PB extrusion to judge the presumptive nucleus state [19,25,29]. However, pronuclear formation may not truly indicate the ploidy of activated oocytes [2].…”

Section: Discussionmentioning

confidence: 99%

“…70(11): 1165-1172, 2008 In bovine, oocyte activation is a major factor for successful production of reconstructed embryos by somatic cell nuclear transfer (SCNT) [34], parthenogenesis and intracytoplasmic sperm injection [23]. During normal fertilization in mammalian species, sperm penetration into the oocyte triggers intracellular Ca 2+ oscillation followed by inactivation of the maturation promoting factor (MPF) composed of the catalytic subunit of cdc2p34 protein kinase and the regulatory subunit cyclin B1/B2 for about 2 hr [37] [29,30], that prevent MPF re-accumulation by reducing cdc2 kinase activity [2] have also been applied. DMAP treatment of metaphase II oocytes induces diploid activation by preventing chromosomal separation and extrusion of the second polar body (PB) [19].…”

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confidence: 99%

“…Although artificial stimuli, such as Ca 2+ ionophore [19], ionomycin [23], ethanol [31], electric pulse [27] and strontium (SrCl 2 ) [21] can activate bovine oocytes by increasing intracellular Ca 2+ , the efficiency of artificial stimuli is limited [25,34]. Therefore, additional chemical agents, such as cycloheximide (CHX, protein phosphorylation inhibitor) [24] and 6-dimethylaminopurine (DMAP, protein synthesis inhibitor) [29,30], that prevent MPF re-accumulation by reducing cdc2 kinase activity [2] have also been applied. DMAP treatment of metaphase II oocytes induces diploid activation by preventing chromosomal separation and extrusion of the second polar body (PB) [19].…”

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Parthenogenetic Development and Ploidy following Various Chemical Activation Regiments of Bovine Oocytes

Ock

Rho

2008

The Journal of Veterinary Medical Science

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ABSTRACT. Bovine oocytes treated using various combinations of 5 µM ionomycin (Ion), 10 µg/ml cycloheximide (CHX) and 5 µg/ml cytochalsin B (CCB) were evaluated to determine developmental rates and ploidy status. The groups were comprised of metaphase II oocytes exposed to Ion for 5 min (Group 1); Ion treatment followed by CHX for 5 hr (Group 2); Ion treatment followed by CHX/CCB for 1 hr and CHX for 4 hr (Group 3); Ion treatment followed by CHX/CCB for 3 hr and CHX for 2 hr (Group 4); and Ion treatment followed by CHX/CCB for 5 hr (Group 5). Group 5 exhibited significantly (P<0.05) lower rates of second polar body (PB) extrusion and higher rates of cleavage and blastocyst development. At 8 hr after Ion treatment, the eggs in groups 2, 3 and 5 were divided into 2 subgroups based on the presence or absence of the second PB and were assessed for cleavage rate and ploidy at the two-cell stage. The cleavage rates did not differ among the activation treatments or between the presence and absence of the second PB in all groups. The diploid rate was significantly (P<0.05) higher in group 5 than in groups 2 and 3. However, the diploid rate in blastocyst-stage parthenotes did not differ among groups 2, 3 and 5. Consequently, oocyte activation by CHX/CCB for 5 hr after Ion treatment could enhance diploid parthenogenetic development in bovine. KEY WORDS: bovine parthenote, chemical activation, ploidy, polar body.J. Vet. Med. Sci. 70(11): 1165-1172, 2008 In bovine, oocyte activation is a major factor for successful production of reconstructed embryos by somatic cell nuclear transfer (SCNT) [34], parthenogenesis and intracytoplasmic sperm injection [23]. During normal fertilization in mammalian species, sperm penetration into the oocyte triggers intracellular Ca 2+ oscillation followed by inactivation of the maturation promoting factor (MPF) composed of the catalytic subunit of cdc2p34 protein kinase and the regulatory subunit cyclin B1/B2 for about 2 hr [37] [29,30], that prevent MPF re-accumulation by reducing cdc2 kinase activity [2] have also been applied. DMAP treatment of metaphase II oocytes induces diploid activation by preventing chromosomal separation and extrusion of the second polar body (PB) [19]. However, CHX does not prevent chromosomal segregation and second PB extrusion, resulting in haploid development. When cytochalasin D (CCD) was added in the medium, chromosomal segregation manifested, but the cytokinesis that induces second PB extrusion was prohibited in CHX-treated oocytes, leading to diploid development with two pronuclei in activated oocytes [4,19]. The combined treatment of ionomycin with DMAP for oocyte activation could induce an abnormal pattern of karyokinesis during the first cell cycle [8], and thus, CHX has widely been used as an alternative approach [19]. Moreover, in cloning, oocyte activation using CHX produces healthy live offspring in cattle [12,17], but the live offspring rate is still low [5].Campbell et al. [3] proposed the hypothesis that when donor cells at G1 stage are reconstructed...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Parthenogenetic Development and Ploidy following Various Chemical Activation Regiments of Bovine Oocytes

Ock

Rho

2008

The Journal of Veterinary Medical Science

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“…Previous experiments using embryonic blastomeres as donor nuclei showed that inefficient oocyte activation of the recipient cytoplast is a major factor in the low efficiency of cloning [8]. Currently, the common methods to activate MII oocytes are a physical stimuli such as an electrical pulse [9], chemical stimuli such as ethanol [10], calcium ionophore [11], ionomycin [12] and inositol 1,4,5-triphosphate [13]. These stimuli were used alone or combined with a protein phosphorylation inhibitor, 6-dimethylaminopurine (6-DMAP) or a protein synthesis inhibitor, cycloheximide.…”

Section: Introductionmentioning

confidence: 99%

Development of swamp buffalo (Bubalus bubalis) embryos after parthenogenetic activation and nuclear transfer using serum fed or starved fetal fibroblasts

et al. 2004

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-The knowledge of oocyte activation and somatic cell nuclear transfer in the swamp buffalo (Bubalus bubalis) is extremely rare. The objectives of this study were the following: (1) to investigate the various activation treatments on the parthenogenetic development of buffalo oocytes, (2) to examine the events of nuclear remodeling and in the in vitro development of cloned buffalo embryos reconstructed with serum fed or starved fetal fibroblats, and (3) to investigate the in vivo development of cloned embryos derived from serum fed or starved cells after transfer into the recipients. The rates of cleavage and blastocyst development were found to be significantly higher (P < 0.05) when the oocytes were activated by the combination treatment of calcium ionophore (A23187) or ethanol followed by 6-DMAP than those activated by electrical pulses and 6-DMAP or other single treatments. Flow cytometric analysis revealed that the percentage in the G0/G1 phase in serum starved cells was significantly (P < 0.05) higher than that in serum fed cells (88.8 ± 6.2 vs. 68.2 ± 2.6). At 1 h post fusion (hpf), most of the transferred nuclei (71%) from serum fed cells did not change in size, and the nuclear envelope remained intact, whereas 29% underwent NEBD and PCC. When serum starved cells were used, 83% of the transferred nuclei underwent NEBD and PCC whereas 17% remained intact. The nuclear swelling and pronucleus (PN) formation were observed at 2-4 and 12 h post activation (hpa), respectively. The remodeled nuclei underwent mitotic division and developed to the 2-cell stage within 18-24 hpa. Fifty-five percent of oocytes reconstructed with serum fed cells were 2PN and 45% were 1PN, whereas 79% of the embryos reconstructed from starved cells were 1PN and 21% were 2PN. The percentage of blastocyst development of the embryos derived from starved cells was higher than that from the serum fed cells (35% vs. 21%, P < 0.05). Pregnancy was detected after the transfer of cloned blastocysts into the recipients but no recipients supported the development to term. The results of this work can be used to establish effective activation protocols for buffalo oocytes which can be used during nuclear transfer experiments.buffalo / oocyte activation / nuclear transfer / nuclear remodeling / somatic cell * Corresponding author: scykt@mahidol.ac.th 66 J. Saikhun et al.

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“…After ETO treatment, however, no spindle is detected and only a single cluster of chromatin can be seen [10]. This [22]. However, experimental evidence demonstrates that this protocol leads to the retention of the PB in normal oocytes [17].…”

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confidence: 92%

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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Inhibition of Protein Kinases after an Induced Calcium Transient Causes Transition of Bovine Oocytes to Embryonic Cycles without Meiotic Completion

Cited by 258 publications

References 0 publications

Parthenogenetic Development and Ploidy following Various Chemical Activation Regiments of Bovine Oocytes

Parthenogenetic Development and Ploidy following Various Chemical Activation Regiments of Bovine Oocytes

Development of swamp buffalo (Bubalus bubalis) embryos after parthenogenetic activation and nuclear transfer using serum fed or starved fetal fibroblasts

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

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