Nuclear transfer in sheep embryos: the effect of cell-cycle coordination between nucleus and cytoplasm and the use of in vitro matured oocytes

Molecular Reproduction and Development
L LiuR M Moor

Abstract

The developmental ability of nuclear transplant sheep embryos derived from in vitro matured oocytes was studied by controlling cell-cycle coordination of donor embryonic nuclei and recipient cytoplasts. Oocytes were recovered from nonatretic antral follicles of adult sheep ovaries and cocultured with follicle shells in M199-based medium supplemented with gonadotrophins in a nonstatic system. Effective activation if IVM oocytes was obtained by applying two pulses of 1.0 kv/cm 22 min apart in inositol-based electroporation medium to oocytes matured in vitro for 27 hr. Synthesis of DNA (S-phase) was assessed by BrdU incorporation and was found to initiate around 5 hpa (hours postactivation) and to persist until 18 hpa. Mitotic blastomeres were induced by treating embryos with 6.6 microM nocodazole for 14-17 hr. Three types of transfers were compared directly: "S-->S," early embryonic nuclei (mostly in S-phase) were transferred to presumptive S-phase cytoplasts; "M-->MII," nocodazole-treated embryonic nuclei (most in M-phase) were transferred to MII-phase cytoplasts; and control (S-->MII), conventional nuclear transfer of fusion and activation simultaneously. The results showed that fusion and recovery rates did not differ among th...Continue Reading

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Citations

Mar 26, 1999·Molecular Reproduction and Development·V ZakhartchenkoG Brem
Aug 24, 2000·Molecular Reproduction and Development·R AlberioV Zakhartchenko
Apr 12, 2002·Cloning and Stem Cells·K H CampbellW A Ritchie
Apr 12, 2002·Cloning and Stem Cells·N KorfiatisO Lacham-Kaplan
Aug 13, 2002·Cloning and Stem Cells·Björn Oback, David Wells
Jan 22, 2004·Cloning and Stem Cells·Liangxue Lai, Randall S Prather
Oct 3, 2006·Cloning and Stem Cells·Stephen SullivanJim McWhir
Nov 14, 2003·Reproductive Biology and Endocrinology : RB&E·Liangxue Lai, Randall S Prather
Oct 20, 2007·Theriogenology·C L Keefer
Feb 14, 2002·Differentiation; Research in Biological Diversity·Karl Illmensee
May 1, 2001·Nature Biotechnology·L Liu
Mar 30, 2001·Human Reproduction·T TakeuchiG D Palermo

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