Hostname: page-component-8448b6f56d-gtxcr Total loading time: 0 Render date: 2024-04-23T20:01:22.968Z Has data issue: false hasContentIssue false
  • English
  • Français

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

Published online by Cambridge University Press:  18 December 2013

Jingling Shen ,
Zhendong Wang ,
Xinghui Shen ,
Zhong Zheng ,
Qinghua Zhang ,
Xiuqing Feng ,
Lili Hu  and
Lei Lei
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: leil086@yahoo.com.cn
Get access Rights & Permissions [Opens in a new window]

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.

Keywords

Enucleated oocytesICSIMouseSCNTβ-Tubulin
Type
Research Article
Information
Zygote , Volume 23 , Issue 1 , February 2015 , pp. 76 - 82
Copyright
Copyright © Cambridge University Press 2013 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Branzini, C., Lavolpe, M., Nodar, F. & Rawe, V.Y. (2007). Visualization of cytoskeleton components during fertilization in mammals. Fertil. Steril. 88, 1435–6.Google Scholar
Brunet, S., Polanski, Z., Verlhac, M.H., Kubiak, J.Z. & Maro, B. (1998). Bipolar meiotic spindle formation without chromatin. Curr. Biol. 8, 1231–4.Google Scholar
Dai, Y., Wang, L., Wang, H., Liu, Y., Li, N., Lyu, Q., Keefe, D.L., Albertini, D.F. & Liu, L. (2006). Fate of centrosomes following somatic cell nuclear transfer (SCNT) in bovine oocytes. Reproduction 131, 1051–61.Google Scholar
Kim, N.H., Chung, K.S. & Day, B.N. (1997). The distribution and requirements of microtubules and microfilaments during fertilization and parthenogenesis in pig oocytes. J. Reprod. Fertil. 111, 143–9.CrossRefGoogle ScholarPubMed
Kwon, D.J., Lee, Y.M., Hwang, I.S., Park, C.K., Yang, B.K. & Cheong, H.T. (2010). Microtubule distribution in somatic cell nuclear transfer bovine embryos following control of nuclear remodeling type. J. Vet. Sci. 11, 93–101.CrossRefGoogle ScholarPubMed
Liu, Z., Schatten, H., Hao, Y., Lai, L., Wax, D., Samuel, M., Zhong, Z-S., Sun, Q-Y. & Prather, R.S. (2006). The nuclear mitotic apparatus (NuMA) protein is contributed by the donor cell nucleus in cloned porcine embryos. Front. Biosci. 1, 1945–57.CrossRefGoogle Scholar
Miki, H., Inoue, K., Ogonuki, N., Mochida, K., Nagashima, H., Baba, T. & Ogura, A. (2004). Cytoplasmic asters are required for progression past the first cell cycle in cloned mouse embryos. Biol. Reprod. 71, 2022–8.Google Scholar
Nagy, A., Gertsenstein, M., Vintersten, K. & Behringer, R.R. (2003). Manipulating the Mouse Embryo: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.Google Scholar
Rybouchkin, A., Heindryckx, B., Van der Elst, J. & Dhont, M. (2002). Developmental potential of cloned mouse embryos reconstructed by a conventional technique of nuclear injection. Reproduction 124, 197–207.CrossRefGoogle ScholarPubMed
Schatten, G., Simerly, C. & Schatten, H. (1985). Microtubule configurations during fertilization, mitosis, and early development in the mouse and the requirement for egg microtubule-mediated motility during mammalian fertilization. Proc. Natl. Acad. Sci. USA 82, 4152–6.CrossRefGoogle ScholarPubMed
Simerly, C., Dominko, T., Navara, C., Payne, C., Capuano, S., Gosman, G., Chong, K.Y., Takahashi, D., Chace, C., Compton, D., Hewitson, L. & Schatten, G. (2003). Molecular correlates of primate nuclear transfer failures. Science 300, 297.Google Scholar
Svarcova, O., Dinnyes, A., Polgar, Z., Bodo, S., Adorjan, M., Meng, Q. & Maddox-Hyttel, P. (2009). Nucleolar re-activation is delayed in mouse embryos cloned from two different cell lines. Mol. Reprod. Dev. 76, 132–41.Google Scholar
Tomioka, I., Mizutani, E., Yoshida, T., Sugawara, A., Inai, K., Sasada, H. & Sato, E. (2007). Spindle formation and microtubule organization during first division in reconstructed rat embryos produced by somatic cell nuclear transfer. J. Reprod. Dev. 53, 835–42.Google Scholar
Van Thuan, N., Wakayama, S., Kishigami, S. & Wakayama, T. (2006). Donor centrosome regulation of initial spindle formation in mouse somatic cell nuclear transfer: roles of gamma-tubulin and nuclear mitotic apparatus protein 1. Biol. Reprod. 74, 777–87.CrossRefGoogle ScholarPubMed
Woods, L.M., Hodges, C.A., Baart, E., Baker, S.M., Liskay, M. & Hunt, P.A. (1999). Chromosomal influence on meiotic spindle assembly: abnormal meiosis I in female Mlh1 mutant mice. J. Cell. Biol. 145, 1395–406.Google Scholar
Yoo, J.G., Demers, S.P., Lian, L. & Smith, L.C. (2007). Developmental arrest and cytoskeletal anomalies of rat embryos reconstructed by somatic cell nuclear transfer. Cloning Stem Cells 9, 382–93.Google Scholar
Yoshida, N. & Perry, A.C. (2007). Piezo-actuated mouse intracytoplasmic sperm injection (ICSI). Nat. Protoc. 2, 296–304.Google Scholar
Zhu, Z.Y., Chen, D.Y., Li, J.S., Lian, L., Lei, L., Han, Z.M. & Sun, Q.Y. (2003). Rotation of meiotic spindle is controlled by microfilaments in mouse oocytes. Biol. Reprod. 68, 943–94.CrossRefGoogle ScholarPubMed