In Vitro Culture of Feline Embryos Increases Stress-induced Heat Shock Protein 70 and Apoptotic Related Genes
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概要
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Developmental competence and quality of in vitro produced embryos has been demonstrated to be lower than in vivo derived embryos. This study aimed specifically to determine the effects of in vitro culture of feline embryos using various culture densities on developmental competence and expression of stress- and apoptotic-related genes in terms of heat shock protein 70 (HSP70) and apoptotic-related (BAX and BCL-2) gene expressions. In experiment 1, we characterized the inducible form of a feline-specific HSP70 mRNA sequence, as it has not been previously reported. The primers for feline HSP70 mRNA were synthesized and tested on heat-treated cat fibroblasts. In experiment 2, feline embryos were cultured at different culture densities (embryo:culture volume; 1:1.25, 1:5 and 1:20). The developmental competence was determined along with HSP70, BAX and BCL-2 transcript abundances using quantitative RT-PCR. In vivo derived embryos were used as a control group. A partial cat HSP70 mRNA sequence (190 bp) was characterized and exhibited high nucleotide identity (93 to 96%) with other species. Cleaved embryos cultured at high density (1:1.25) developed to blastocysts at a lower rate than those generated from lower densities. Irrespective of the culture densities used, in vitro cultured blastocysts showed increased levels of HSP70 and BAX transcripts compared with in vivo counterparts. Blastocysts derived from the highest culture density (1:1.25) showed higher levels of upregulation of HSP70 and BAX transcripts than those cultured at lower culture densities (1:5 and 1:20). In conclusion, increased levels of pro-apoptotic (BAX) and stress-response (HSP70) transcripts correlated with developmental incompetence of embryos cultured at high embryonic density, indicating that stress accumulated during in vitro embryo culture affected the fate for embryo development and quality.
- 日本繁殖生物学会の論文
著者
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Techakumphu Mongkol
Department Of Obstetrics Gynaecology And Reproduction Faculty Of Veterinary Science Chulalongkorn Un
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PHUTIKANIT Nawapen
Department of Obstetrics Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn
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THARASANIT Theerawat
Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University
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PHUTIKANIT Nawapen
Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand
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SANANMUANG Thanida
Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand
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NGUYEN Catherine
Laboratoire INSERM U928 - TAGC (Technologies Avancées pour le Génome et la Clinique), Parc Scientifique et Technologique de Luminy, Marseille, France
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MANEE-IN Sukanya
Faculty of Veterinary Science, Mahidol University, Nakorn-pathom, Thailand
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