Bovine parthenogenesis is characterized by abnormal chromosomal complements: Implications for maternal and paternal co-dependence during early bovine development

Winger, Q. A.; Fuente, Rabindranath De La; King, W. A.; Armstrong, David T.; Watson, Andrew J.

doi:10.1002/(sici)1520-6408(1997)21:2<160::aid-dvg5>3.0.co;2-5

Cited by 55 publications

(46 citation statements)

References 52 publications

(79 reference statements)

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“…The results indicate that prolonged activation with CHX in combination with CCB increased the number of diploid parthenotes while reducing the number of haploid parthenotes. In addition, activation with CHX alone after Ion treatment appeared to induce a high diploid rate, as reported previously by Winger et al [36]. From the above results, it can be suggested that although CCB prevented extrusion of the second PB, treatment for 1 or 3 hr could not completely block extrusion.…”

Section: Discussionsupporting

confidence: 84%

“…This may be because a nonspecific chemical agent like CHX affects several metabolic pathways and induces chromosomal abnormalities by preventing PB extrusion and premature DNA synthesis [2]. Although a few reports are available on ploidy analysis of both early and late stage parthenogenic embryos in bovine [36], there is a lack of information on ploidy status following activation by CHX with or without CCB. The present study did not directly compare early and late stage parthenogenic embryos, but the blastocysts obtained by CHX with or without CCB treatment on day 9 showed extremely lower rates of haploidy and increased rates of mixoploidy compared with the two-cell parthenotes.…”

Section: Discussionmentioning

confidence: 99%

“…We presume that haploid parthenotes often undergo developmental arrest at an early stage. On the other hand, diploid parthenotes might possess developmental competency similar to IVF embryos [18,36], and hence, they could develop to the blastocyst stage. All blastocysts developed from eggs treated with CCB exhibited mixoploidy, and this phenomenon has often been observed in IVF embryos.…”

Section: Discussionmentioning

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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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: Discussionsupporting

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

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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“…In 280 addition, we have a similar result after parthenogenetic activation of sham-injected oocytes with ionomycin and ethanol (data not shown). Chemicals such as ethanol, ionomycin or calcium ionophore combined with or without CHX were not sufficient to induce parthenogenetic development in bovine oocytes [42,43]. Induction of diploidization by combining the calcium oscillation-inducing activators with cytochalasin or 6-DMAP was necessary to improve the 285 parthenogenetic development of bovine oocytes [24,44,45].…”

Section: Discussionmentioning

confidence: 99%

A combined treatment of ionomycin with ethanol improves blastocyst development of bovine oocytes harvested from stored ovaries and microinjected with spermatozoa

et al. 2009

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Regardless of the presence of sperm-borne oocyte-activating factors, activation of bovine oocytes with exogenous activation stimuli is required for further development after intracytoplasmic sperm injection (ICSI). The present study was designed to develop a new activation regimen for improving the blastocyst yield after ICSI of bovine oocytes harvested from ovaries stored at 10-12 oC for 24 h. Following ICSI, oocytes were treated with 5 μM ionomycin for 5 min, 7% ethanol for 5 or 10 min, ionomycin followed by ethanol (5 or 10 min), ionomycin followed by 10 µg/mL Cycloheximide for 5 h, or ionomycin followed by 1.9 mM 6-dimethylaminopurine for 3 h. Across the activation regimens, the cleavage rates of ICSI oocytes (45-77%) were higher than those of parthenogenetically activated oocytes (11-21%; P<0.05). Activating the ICSI oocytes with ionomycin plus ethanol improved the blastocyst yield (29-30%) comparing to the non-treated oocytes (12%; P<0.05) but the other regimens did not (9-18%; P>0.05). The higher blastocyst yields were due to increasing the proportion of ICSI oocytes that passed through the early postfertilization events until cleavage. None of the regimens have any adverse effect on the quality of the blastocysts regarding the total cell number or the proportion of the inner cell mass cells. Thus, a new activation regimen composed from two triggers for single calcium increase has been proven effective to improve the blastocyst yield after bovine ICSI using oocytes harvested from stored ovaries.Dear Dr. John P. Kastelic, Thank you very much for prompt reply and accepting our manuscript for publication in Theriogenology. We have replied for each comment from Co-editor and the reviewer-2 in pointby-point basis.Sincerely yours, Shinichi Hochi (Corresponding author)Hany Abdalla (First author)Co-editor 1-P value has been added to the Abstract (Lines 27,29).2-The number of the references in Introduction section has been reduced. Reviewer #21-Regarding omitting the two groups (5 min ethanol group and ionomycin followed by 5 min ethanol group); We do not think that the presence of these two groups causes any difficulty to recognize the superiority of ionomycin and ethanol combination. Since activation of 7% ethanol for 5 min 4 hr after ICSI is a commonly applied method for bovine ICSI, presence of such group must be important to declare the superiority of the new combination (ionomycin and ethanol) over the methods of ethanol alone under the same experimental circumstance regarding oocyte quality, injection skill, culture condition, and so on. For the ionomycin followed by 5 min ethanol, the efficiency of this method to improve the cleavage and the blastocyst yield was similar to that of ionomycin followed by 10 min ethanol, making it possible to choose the method in which oocytes are exposed to minimum exogenous stimuli. Tables 1 and 2. 2-The time of 2 nd polar body detection has been added in footnote of 15Running head: Bovine oocyte activation after ICSI * Manuscript 2 AbstractRegardless of the pres...

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“…For example, the use of cycloheximide (CHX) during activation of bovine SCNTembryos was associated with delayed DNA synthesis (Alberio et al 2001b), and the use of 6-dimethylaminopurine (DMAP) as the activating agent often resulted in a high proportion of aneuploid embryos (Winger et al 1997, Van De Velde et al 1999, Bhak et al 2006.…”

Section: Introductionmentioning

confidence: 99%

Activation of bovine somatic cell nuclear transfer embryos by PLCZ cRNA injection

Ross¹,

Rodríguez²,

Iager³

et al. 2009

Reproduction

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The production of cloned animals by the transfer of a differentiated somatic cell into an enucleated oocyte circumvents fertilization. During fertilization, the sperm delivers a sperm-specific phospholipase C (PLCZ) that is responsible for triggering Ca 2C oscillations and oocyte activation. During bovine somatic cell nuclear transfer (SCNT), oocyte activation is artificially achieved by combined chemical treatments that induce a monotonic rise in intracellular Ca 2C and inhibit either phosphorylation or protein synthesis. In this study, we tested the hypothesis that activation of bovine nuclear transfer embryos by PLCZ improves nuclear reprogramming. Injection of PLCZ cRNA into bovine SCNTunits induced Ca 2C oscillations similar to those observed after fertilization and supported high rates of blastocyst development similar to that seen in embryos produced by IVF. Furthermore, gene expression analysis at the eight-cell and blastocyst stages revealed a similar expression pattern for a number of genes in both groups of embryos. Lastly, levels of trimethylated lysine 27 at histone H3 in blastocysts were higher in bovine nuclear transfer embryos activated using cycloheximide and 6-dimethylaminopurine (DMAP) than in those activated using PLCZ or derived from IVF. These results demonstrate that exogenous PLCZ can be used to activate bovine SCNT-derived embryos and support the hypothesis that a fertilization-like activation response can enhance some aspects of nuclear reprogramming.

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Bovine parthenogenesis is characterized by abnormal chromosomal complements: Implications for maternal and paternal co-dependence during early bovine development

Cited by 55 publications

References 52 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

A combined treatment of ionomycin with ethanol improves blastocyst development of bovine oocytes harvested from stored ovaries and microinjected with spermatozoa

Activation of bovine somatic cell nuclear transfer embryos by PLCZ cRNA injection

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