Effect of Osmolarity of Culture Medium on Imprinting and Apoptotic Gene Expression in Miniature Pig Nuclear Transfer Embryos

서지정보

· 발행기관 : 한국동물번식학회
· 수록지 정보 : Reproductive & developmental biology / 32권 / 3호
· 저자명 : Mi-Rung Park, In-Sun Hwang, Joo-Hyun Shim, Hyo-Jin Moon, Dong-Hoon Kim, Yeoung-Kyu Ko, Hwan-Hoo Seong, Gi-Sun Im

목차

ABSTRACT INTRODUCTION MATERIALS AND METHODS Oocyte Recovery and Maturation Preparation of Nuclear Donor Cells Production of Nuclear Transfer Embryos Terminal DeoxynucleotidyI Transferase-Mediated dUTP Nick-End Labeling (TUNEL) Assay Collection of In Vivo Blastocysts Realtime PCR Quantification Statistical Analysis RESULTS DISCUSSION REFERENCES

영어초록

This study was conducted to investigate the development and gene expression in miniature pig nuclear transfer (mNT) embryos produced under different osmolarity culture conditions. Control group of mNT embryos was cultured in PZM-3 for 6 days. Treatment group of mNT embryos was cultured in modified PZM-3 with NaCl (mPZM-3, 320 mOsmol) for 2 days, and then cultured in PZM-3 (270 mOsmol) for 4 days. Blastocyst formation rate of the treatment group was significantly higher than the control and the apoptosis rate was significantly lower in treatment group. Bax- and caspase-3 mRNA expression were significantly higher in the control than the treatment group. Also, the majority of imprinting genes were expressed aberrantly in in vitro produced mNT blastocysts compared to in vivo derived blastocyst H19 and Xist mRNA expression were significantly lower in the control than the treatment group or in vivo. IGF2 mRNA expression was significantly higher in the control than the treatment group or in vivo. IGF2r mRNA expression was significantly lower in the control. Methylation profiles of individual DNA strands in H19 upstream T-DMR sequences showed a similar methylation status between treatment group and in vivo. These results indicate that the modification of osmolarity in culture medium at early culture stage could provide more beneficial culture environments for mNT embryos.

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