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Cytochalasin B treatment of mouse oocytes during intracytoplasmic sperm injection (ICSI) increases embryo survival without impairment of development

Published online by Cambridge University Press:  15 August 2011

Li-li Hu ,
Xing-hui Shen ,
Zhong Zheng ,
Zhen-dong Wang ,
Zhong-hua Liu ,
Lian-hong Jin  and
Lei Lei
Li-li Hu
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, 150081, Harbin, China.
Xing-hui Shen
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, 150081, Harbin, China.
Zhong Zheng
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, 150081, Harbin, China.
Zhen-dong Wang
Affiliation:
College of Life Science, Northeast Agricultural University, No. 59 Mucai Street Xiangfang District, 150030, Harbin, China.
Zhong-hua Liu
Affiliation:
College of Life Science, Northeast Agricultural University, No. 59 Mucai Street Xiangfang District, 150030, Harbin, China.
Lian-hong Jin*
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin 150081, China.
Lei Lei*
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin 150081, China.
*
All correspondence to: Lian-hong Jin or Lei Lei. Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin 150081, China. Tel: +86 451 86674518. Fax: +86 451 87503325. E-mail: wstjlh@126.com or leil086@yahoo.com.cn.
All correspondence to: Lian-hong Jin or Lei Lei. Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin 150081, China. Tel: +86 451 86674518. Fax: +86 451 87503325. E-mail: wstjlh@126.com or leil086@yahoo.com.cn.
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Summary

Intracytoplasmic sperm injection (ICSI) is a technique commonly used in clinical and research settings. In mouse oocytes, conventional ICSI has a poor survival rate caused by a high level of lysis. Cytochalasin B (CB) is a toxic microfilament-inhibiting agent that is known to relax the cytoskeleton and enhance the flexibility of oocytes. CB has been used widely in nuclear transfer experiments to improve the success rate of the micromanipulation, however information describing the use of CB in ICSI is limited. Here, we demonstrated that the addition of 5 μg/ml CB to the manipulation medium of ICSI procedure significantly improved the survival rate of the ICSI embryos (80.74% vs. 89.50%, p < 0.05), and that there was no harm for the in vitro or in vivo development. The birth rates and birth weights were not significantly different between the CB-treated and -untreated groups. Interestingly, the microfilaments of the ICSI embryos were almost undetectable immediately after CB treatment; however, they gradually re-appeared and had fully recovered to the normal level 2 h later. Moreover, CB did not disturb spindle rotation, second polar body formation or pronuclei migration, and had no effect on the microtubules. We thus conclude that ICSI manipulation in CB-containing medium results in significantly improved survival rate of mouse ICSI embryos, and that short-term treatment with CB during ICSI manipulation does not have adverse effects on the development of ICSI embryos.

Keywords

Cytochalasin BIntracytoplasmic sperm injectionMicrofilamentMicrotubuleMouse embryo
Type
Research Article
Information
Zygote , Volume 20 , Issue 4 , November 2012 , pp. 361 - 369
Copyright
Copyright © Cambridge University Press 2011

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