Assessing Chromosomal Abnormalities in Two-Cell Bovine In Vitro-Fertilized Embryos by Using Fluorescent In Situ Hybridization with Three Different Cloned Probes1

Slimane, Wafa; Heyman, Yvan; Lavergne, Y.; Humblot, P.; Renard, Jean Paul

doi:10.1095/biolreprod62.3.628

Cited by 37 publications

(23 citation statements)

References 24 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].…”

Section: Discussionmentioning

confidence: 99%

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%

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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“…A haploid/diploid blastocyst was observed in the SCM-CO culture group. Mixoploidy of 1n/2n is usually found in 2-cell (Slimane et al 2000) and in 4-to 32-cell embryos (Iwasaki et al 1992). It has been suggested that the 1n/2n mosaic results from the retained activity of a polar body or is a product of polyspermy (King et al 1981, Iwasaki et al 1992.…”

Section: Discussionmentioning

confidence: 99%

Conditioned medium increases the polyploid cell composition of bovine somatic cell nuclear-transferred blastocysts

White

Aston

et al. 2004

Reproduction

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The effects of bovine cumulus cell-conditioned medium on cloned bovine embryonic development and subsequent chromosome complement were examined using an air-dry procedure. Conditioned media were prepared using CR1aa supplemented with either fetal bovine serum (FBS) or bovine serum albumin (BSA). Nuclear-transferred embryos were reconstructed with nuclei from cumulus cells. Similar cleavage, morula, and blastocyst development was observed in conditioned media groups compared with the co-culture group. No differences (P > 0.05) were observed in the composition of blastocyst chromosomes after co-culture in different media, either with or without starvation of donor cells. The overall diploid blastocyst rate ranged from 75% to 84%. Chromosomal complement of blastocysts, however, was very different between conditioned medium and co-culture treatments. Overall incidence of chromosomal anomalies was 40% in conditioned medium, which was significantly higher (P < 0.001) than the co-culture group (20%). Moreover, a higher incidence (P < 0.05) of chromosomally abnormal blastocysts (41.5%) was observed after culture with FBS-containing conditioned medium than those cultured in BSA-containing conditioned medium (31.4%). No diploid improvement was observed after exchange of the culture system from conditioned medium to co-culture, or from co-culture to conditioned medium after the first 72 h of culture. The results of this study also indicated that the overall cell number was much lower (P < 0.01) in blastocysts with chromosomal abnormalities than those with a normal diploid state. We have concluded that medium conditioned with bovine cumulus cells increases the incidence of chromosomal anomalies in nuclear reconstructed embryos.

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“…These differences include darker cytoplasm, lower density [64], more lipids, specifically, more triglycerides and less lipids from other classes [1], swollen blastomeres [81], a more fragile zona pellucida [19], differences in intercellular communication [10], and a higher incidence of chromosomal abnormalities [72,82]. All of these factors may contribute to the higher sensitivity to cryoinjury exhibited by IVP embryos.…”

Section: Differences Between In Vivo-and In Vitro-produced Embryosmentioning

confidence: 99%

Factors influencing oocyte and embryo quality in cattle

et al. 2001

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Assessing Chromosomal Abnormalities in Two-Cell Bovine In Vitro-Fertilized Embryos by Using Fluorescent In Situ Hybridization with Three Different Cloned Probes1

Cited by 37 publications

References 24 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

Conditioned medium increases the polyploid cell composition of bovine somatic cell nuclear-transferred blastocysts

Factors influencing oocyte and embryo quality in cattle

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