title: Piezo-assisted nuclear transfer affects cloning efficiency and may cause apoptosis.
abstract: Even though it generates healthy adults, nuclear transfer in mammals remains an inefficient process. Mainly attributed to abnormal reprograming of the donor chromatin, this inefficiency may also be caused at least partly by a specific effect of the cloning technique which has not yet been well investigated. There are two main procedures for transferring nuclei into enucleated oocytes: fusion and piezoelectric microinjection, the latter being used mostly in mice. We have, therefore, decided to compare the quality and the developmental ability, both in vivo and in vitro, of embryos reconstructed with electrofusion or piezoelectric injection. In addition, the effect of piezo setups of differing electric strengths was investigated. Along with the record of the rate of development, we compared the nuclear integrity in the blastomeres during the first cleavages as well as the morphological and cellular quality of the blastocysts. Our results show that the piezo-assisted micromanipulation can induce DNA damage in the reconstructed embryos, apoptosis, and reduced cell numbers in blastocysts as well as a lower rate of development to term. Even if piezo-driven injection facilitates a faster and more efficient rate of reconstruction, it should be used with precaution and with as low parameters as possible.
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yu
authors.forename: Chenhui
authors.initials: C
authors.lastname: Ding
authors.forename: Eryao
authors.initials: E
authors.lastname: Wang
authors.forename: Xinjie
authors.initials: X
authors.lastname: Chen
authors.forename: Xuemei
authors.initials: X
authors.lastname: Li
authors.forename: Chunli
authors.initials: C
authors.lastname: Zhao
authors.forename: Yong
authors.initials: Y
authors.lastname: Fan
authors.forename: Liu
authors.initials: L
authors.lastname: Wang
authors.forename: Nathalie
authors.initials: N
authors.lastname: Beaujean
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhou
authors.forename: Alice
authors.initials: A
authors.lastname: Jouneau
authors.forename: Weizhi
authors.initials: W
authors.lastname: Ji
mesh_terms: D000818:Animals; D017209:Apoptosis; D001755:Blastocyst / Q000166:cytology; D019976:Cloning, Organism / Q000379:methods*; D004624:Embryo Transfer; D005260:Female; D005455:Fluorescent Antibody Technique; D020287:In Situ Nick-End Labeling; D051379:Mice; D008815:Mice, Inbred Strains; D008845:Microinjections / Q000379:methods; D018613:Microscopy, Confocal; D053652:Nuclear Transfer Techniques; D011247:Pregnancy; D011256:Pregnancy Outcome; D020554:Sperm Injections, Intracytoplasmic