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The effect of the time interval between injection and parthenogenetic activation on the spindle formation and the in vitro developmental potential of somatic cell nuclear-transferred rat oocytes

Published online by Cambridge University Press:  14 August 2009

Shigetoshi Mizumoto ,
Yoko Kato  and
Yukio Tsunoda
Shigetoshi Mizumoto
Affiliation:
Laboratory of Animal Reproduction, College of Agriculture, Kinki University, Nara, 631–8505, Japan.
Yoko Kato
Affiliation:
Laboratory of Animal Reproduction, College of Agriculture, Kinki University, Nara, 631–8505, Japan.
Yukio Tsunoda*
Affiliation:
Laboratory of Animal Reproduction, College of Agriculture, Kinki University, Nara, 631–8505, Japan. Laboratory of Animal Reproduction, College of Agriculture, Kinki University, Nara, 631–8505, Japan.
*
All correspondence to: Yukio Tsunoda. Laboratory of Animal Reproduction, College of Agriculture, Kinki University, Nara, 631–8505, Japan. Tel +81 742 43 5143. Fax: +81 742 43 5393. e-mail: tsunoda@nara.kindai.ac.jp
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Summary

We examined the optimal conditions for somatic cell nuclear transfer (SCNT) in the rat. First, we examined the effect of preincubation time before activation on SCNT rat oocytes produced in the presence of MG132 with regard to spindle formation and the potential to develop into blastocysts. The spindles of SCNT oocytes continued to elongate with an increase in the culture duration and, in approximately half of oocytes, the chromosomes were distributed along the spindles at 120 min after incubation. Such abnormal spindle formation in SCNT oocytes is a possible reason for the low developmental potential of SCNT rat oocytes. To inhibit the formation of abnormal spindle formation, we examined secondly the developmental potential of rat SCNT oocytes that had been preincubated with nocodazole and demecolcine instead of MG132. The developmental rates in SCNT oocytes, however, were decreased. For successful rat somatic cell cloning, two steps might be required: (1) to culture the somatic cell nuclei for a sufficient time in MII oocyte cytoplasm to enhance nuclear reprogramming; and (2) to induce normal spindle formation with normal chromosomal construction.

Keywords

DemecolcineMG132NocodazoleNuclear transferParthenogenesisTSA
Type
Research Article
Information
Zygote , Volume 18 , Issue 1 , February 2010 , pp. 9 - 15
Copyright
Copyright © Cambridge University Press 2009

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