Parthenogenetic activation and somatic cell nuclear transfer of porcine oocytes activated by an electric pulse and AZD5438 treatment

We examined the in vitro developmental competence of parthenogenetic activation (PA) oocytes activated by an electric pulse (EP) and treated with various concentrations of AZD5438 for 4 h. Treatment with 10 µM AZD5438 for 4 h significantly improved the blastocyst formation rate of PA oocytes in comp...

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Bibliographic Details
Published in:Zygote (Cambridge) 2017-08, Vol.25 (4), p.453-461
Main Authors: Li, Xiao-Chen, Guo, Qing, Zhu, Hai-Ying, Jin, Long, Zhang, Yu-Chen, Zhang, Guang-Lei, Xing, Xiao-Xu, Xuan, Mei-Fu, Luo, Qi-Rong, Luo, Zhao-Bo, Wang, Jun-Xia, Cui, Cheng-Du, Li, Wen-Xue, Cui, Zheng-Yun, Yin, Xi-Jun, Kang, Jin-Dan
Format: Article
Language:English
Subjects:
Adenine - analogs & derivatives
Adenine - pharmacology
Animals
Apoptosis
Apoptosis - drug effects
Apoptosis - genetics
Bcl-2 protein
Blastocyst - physiology
Blastocysts
Cell activation
Cell division
Chromosomes
Electric Stimulation
Embryos
Female
Gene expression
Gene Expression Regulation, Developmental
Genes
GPI-Linked Proteins - metabolism
Imidazoles - administration & dosage
Imidazoles - pharmacology
In vitro methods and tests
In Vitro Oocyte Maturation Techniques - methods
Karyotyping
Kinases
Laboratories
Maturation
Maturation-promoting factor
Nuclear transfer
Nuclear Transfer Techniques
Oct-4 protein
Oocytes
Oocytes - drug effects
Oocytes - physiology
Parthenogenesis - drug effects
Parthenogenesis - physiology
Phosphorylation
Pluripotency
Polymerase chain reaction
Polyvinyl alcohol
Proteins
Pyrimidines - administration & dosage
Pyrimidines - pharmacology
Rodents
Somatic cell nuclear transfer
Stem cells
Studies
Swine
Transgenic animals
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Summary:We examined the in vitro developmental competence of parthenogenetic activation (PA) oocytes activated by an electric pulse (EP) and treated with various concentrations of AZD5438 for 4 h. Treatment with 10 µM AZD5438 for 4 h significantly improved the blastocyst formation rate of PA oocytes in comparison with 0, 20, or 50 µM AZD5438 treatment (46.4% vs. 34.5%, 32.3%, and 24.0%, respectively; P < 0.05). The blastocyst formation rate was higher in the group treated with AZD5438 for 4 h than in the groups treated with AZD5438 for 2 or 6 h (42.8% vs. 38.6% and 37.2%, respectively; P > 0.05). Furthermore, 66.67% of blastocysts derived from these AZD5438-treated PA oocytes had a diploid karyotype. The blastocyst formation rate of PA and somatic cell nuclear transfer (SCNT) embryos was similar between oocytes activated by an EP and treated with 2 mM 6-dimethylaminopurine for 4 h and those activated by an EP and treated with 10 µM AZD5438 for 4 h (11.11% vs. 13.40%, P > 0.05). In addition, the level of maturation-promoting factor (MPF) was significantly decreased in oocytes activated by an EP and treated with 10 µM AZD5438 for 4 h. Finally, the mRNA expression levels of apoptosis-related genes (Bax and Bcl-2) and pluripotency-related genes (Oct4, Nanog, and Sox2) were checked by RT-PCR; however, there were no differences between the AZD5438-treated and non-treated control groups. Our results demonstrate that porcine oocyte activation via an EP in combination with AZD5438 treatment can lead to a high blastocyst formation rate in PA and SCNT experiments.
ISSN:0967-1994
1469-8730
DOI:10.1017/S0967199417000272