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Abnormal dynamic changes in β-tubulin in somatic nuclear transfer cloned mouse embryos

Published online by Cambridge University Press:  18 December 2013

Jingling Shen
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin, Heilongjiang Province, 150081, China.
Zhendong Wang
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin, Heilongjiang Province, 150081, China.
Xinghui Shen
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin, Heilongjiang Province, 150081, China.
Zhong Zheng
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin, Heilongjiang Province, 150081, China.
Qinghua Zhang
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin, Heilongjiang Province, 150081, China.
Xiuqing Feng
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin, Heilongjiang Province, 150081, China.
Lili Hu
Affiliation:
Department of Histology and Embryology, Harbin Medical University, 194 Xuefu Road, Nangang District, Harbin, Heilongjiang Province, 150081, China.
Lei Lei*
Affiliation:
Department of Histology and Embryology, 194 Xuefu Road, Nangang District, Harbin, Heilongjiang Province, 150081, China.
*
All correspondence to: Lei Lei. Department of Histology and Embryology, 194 Xuefu Road, Nangang District, Harbin, Heilongjiang Province, 150081, China. Tel: +86 0451 86674518. Fax: +86 0451 87503325. e-mail: [email protected]

Summary

The efficiency of somatic cell nuclear transfer (SCNT) cloning remains low, thus limiting the applications of this technique. In this study, we used immunochemistry and confocal microscopy to detect the microtubule component, β-tubulin, in SCNT, parthenogenetic (PA), and intracytoplasmic sperm injection (ICSI) embryos before the first mitotic division. β-Tubulin is the component subunit of microtubule, which plays critical roles in regulating localization of cellular organelles, and the growth, maturation and fertilization of oocytes. Our results demonstrated similar changes of spindle patterns in PA and ICSI embryos. The second meiotic division resumed 1 h post-treatment, and the cytoplasmic asters (CAs) disappeared. After about 4–6 h of treatment, pronuclei formed with the midbodies connecting each other. Meanwhile, the CAs reappeared and a microtubule network developed in the cytoplasm. However, SCNT embryos showed abnormal multipolar spindles, and the pseudopronuclei that contained many nucleoli existed after 6 h of SrCl2 activation. Enucleated oocytes alone did not form spindle-like structures when they were artificially activated for 6 h, indicating that somatic cell chromosomes might be necessary for spindle formation in SCNT embryos. These results demonstrated abnormal changes of β-tubulin in mouse SCNT embryos, compared with PA and ICSI embryos.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2013 

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