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Increase in DNA fragmentation and apoptosis-related gene expression in frozen-thawed bovine blastocysts

Published online by Cambridge University Press:  01 May 2006

Sae Young Park
Affiliation:
Maria Infertility Hospital Medical Institute/Maria Biotech, Seoul 130-812, Korea.
Eun Young Kim
Affiliation:
Maria Infertility Hospital Medical Institute/Maria Biotech, Seoul 130-812, Korea.
Xiang Shun Cui
Affiliation:
Chungbuk National University, Gaesin-dong, Cheongju, Chungbuk, Korea.
Jin Cheol Tae
Affiliation:
Maria Infertility Hospital Medical Institute/Maria Biotech, Seoul 130-812, Korea.
Won Don Lee
Affiliation:
Maria Infertility Hospital, Seoul 130-812, Korea.
Nam Hyung Kim
Affiliation:
Chungbuk National University, Gaesin-dong, Cheongju, Chungbuk, Korea.
Se Pill Park*
Affiliation:
Maria Infertility Hospital Medical Institute/Maria Biotech, Seoul 130-812, Korea.
Jin Ho Lim
Affiliation:
Maria Infertility Hospital, Seoul 130-812, Korea.
*
All correspondence to: Se Pill Park, PhD, Department of Animal Sciences, Maria Infertility Hospital Medical Institute/Maria Biotech, Sinseol-Dong, Dongdeamun-Gu, Seoul 130-812. e-mail: sppark@mariababy.com

Summary

Evaluation of apoptosis and expression level of apoptosis-related genes is useful for examining the variation in embryo quality according to environmental change. The objective of this study was to investigate DNA fragmentation and apoptosis-related gene expression patterns in frozen-thawed bovine blastocysts. In vitro produced day 7 blastocysts were frozen by two different vitrification methods (conventional 0.25 ml straw or MVC straw). After thawing, DNA fragmentation of surviving embryos was examined by TUNEL assay, and the expression patterns of their apoptotic genes (survivin, Fas, Hsp 70 and caspase-3) were evaluated using real-time quantitative reverse transcriptase polymerase chain reaction. In vitro survival rates of frozen-thawed embryos were higher following the MVC vitrification method (88.2% re-expanded at 24 h, 77.1% hatching at 48 h) than the conventional (C) vitrification method (77.0% re-expanded at 24 h, 66.7% hatching at 48 h). However, both vitrified methods resulted in a significantly higher apoptotic index (C vitrification method 11.9%, MVC vitrification method 11.0%) than in non-frozen embryos (3.0%). Expression levels of survivin, Fas, caspase-3, and Hsp 70 were also increased in the frozen-thawed embryos compared with non-frozen embryos. These results indicate that the cryopreservation procedure might cause damage that results in an increase in DNA fragmentation and apoptosis-related gene transcription, reducing developmental capacity of frozen-thawed embryos.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2006

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