Assessment of nuclear transfer techniques to prevent the transmission of heritable mitochondrial disorders without compromising embryonic development competence in mice

Jitesh Neupane; Mado Vandewoestyne; Sabitri Ghimire; Yuechao Lu; Chen Qian; Rudy Van Coster; Jan Gerris; Tom Deroo; Dieter Deforce; Petra De Sutter; Björn Heindryckx

doi:10.1016/j.mito.2014.09.003

Assessment of nuclear transfer techniques to prevent the transmission of heritable mitochondrial disorders without compromising embryonic development competence in mice

Mitochondrion. 2014 Sep:18:27-33. doi: 10.1016/j.mito.2014.09.003. Epub 2014 Sep 16.

Authors

Jitesh Neupane¹, Mado Vandewoestyne², Sabitri Ghimire³, Yuechao Lu⁴, Chen Qian⁵, Rudy Van Coster⁶, Jan Gerris⁷, Tom Deroo⁸, Dieter Deforce⁹, Petra De Sutter¹⁰, Björn Heindryckx¹¹

Affiliations

¹ Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium. Electronic address: [email protected].
² Laboratory for Pharmaceutical Biotechnology, Ghent University, Ghent, Belgium. Electronic address: [email protected].
³ Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium. Electronic address: [email protected].
⁴ Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium. Electronic address: [email protected].
⁵ Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium. Electronic address: [email protected].
⁶ Department of Pediatric Neurology and Metabolism, Ghent University Hospital, Ghent, Belgium. Electronic address: [email protected].
⁷ Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium. Electronic address: [email protected].
⁸ Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium. Electronic address: [email protected].
⁹ Laboratory for Pharmaceutical Biotechnology, Ghent University, Ghent, Belgium. Electronic address: [email protected].
¹⁰ Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium. Electronic address: [email protected].
¹¹ Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium. Electronic address: [email protected].

PMID: 25229667
DOI: 10.1016/j.mito.2014.09.003

Abstract

To evaluate and compare mitochondrial DNA (mtDNA) carry-over and embryonic development potential between different nuclear transfer techniques we performed germinal vesicle nuclear transfer (GV NT), metaphase-II spindle-chromosome-complex (MII-SCC) transfer and pronuclear transfer (PNT) in mice. No detectable mtDNA carry-over was seen in most of the reconstructed oocytes and embryos. No significant differences were seen in mtDNA carry-over rate between GV NT (n=20), MII-SCC transfer (0.29 ± 0.63; n=21) and PNT (0.29 ± 0.75; n=25). Blastocyst formation was not compromised after either PNT (88%; n=18) or MII-SCC transfer (86%; n=27). Further analysis of blastomeres from cleaving embryos (n=8) demonstrated undetectable mtDNA carry-over in all but one blastomere. We show that NT in the germ line is potent to prevent transmission of heritable mtDNA disorders with the applicability for patients attempting reproduction.

Keywords: Embryonic development potential; Mitochondrial DNA (mtDNA); Nuclear transfer (NT); mtDNA carry-over.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Blastocyst / physiology
Blastomeres / chemistry
DNA, Mitochondrial / analysis
Embryonic Development*
Female
Male
Mice
Mitochondrial Diseases / prevention & control*
Nuclear Transfer Techniques / adverse effects*
Pregnancy

Substances

DNA, Mitochondrial