Research article summary (published 19 Dec 2007):

Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer.

Full Abstract

BACKGROUND: The mitochondrial DNA (mtDNA) of the cloned sheep "Dolly" and nine other ovine clones produced by somatic cell nuclear transfer (SCNT) was reported to consist only of recipient oocyte mtDNA without any detectable mtDNA contribution from the nucleus donor cell. In cattle, mouse and pig several or most of the clones showed transmission of nuclear donor mtDNA resulting in mitochondrial heteroplasmy. To clarify the discrepant transmission pattern of donor mtDNA in sheep clones we analysed the mtDNA composition of seven fetuses and five lambs cloned from fetal fibroblasts. RESULTS: The three fetal fibroblast donor cells used for SCNT harboured low mtDNA copy numbers per cell (A: 753 +/- 54, B: 292 +/- 33 and C: 561 +/- 88). The ratio of donor to recipient oocyte mtDNAs was determined using a quantitative amplification refractory mutation system (ARMS) PCR (i.e. ARMS-qPCR). For quantification of SNP variants with frequencies below 0.1% we developed a restriction endonuclease-mediated selective quantitative PCR (REMS-qPCR). We report the first cases (n = 4 fetuses, n = 3 lambs) of recipient oocyte/nuclear donor mtDNA heteroplasmy in SCNT-derived ovine clones demonstrating that there is no species-effect hindering ovine nucleus-donor mtDNA from being transmitted to the somatic clonal offspring. Most of the heteroplasmic clones exhibited low-level heteroplasmy (0.1% to 0.9%, n = 6) indicating neutral transmission of parental mtDNAs. High-level heteroplasmy (6.8% to 46.5%) was observed in one case. This clone possessed a divergent recipient oocyte-derived mtDNA genotype with three rare amino acid changes compared to the donor including one substitution at an evolutionary conserved site. CONCLUSION: Our study using state-of-the-art techniques for mtDNA quantification, like ARMS-qPCR and the novel REMS-qPCR, documents for the first time the transmission of donor mtDNA into somatic sheep clones. MtDNA heteroplasmy was detected in seven of 12 clones tested, whereby all but one case revealed less than 1% mtDNA contribution from the nuclear donor cell suggesting neutral segregation.

Author information

Author/s: Burgstaller, Jörg P (JP); Schinogl, Pamela (P); Dinnyes, Andras (A); Müller, Mathias (M); Steinborn, Ralf (R);

Affiliation: Institute of Animal Breeding and Genetics, University of Veterinary Medicine, Veterinaerplatz 1, 1210 Vienna, Austria. joerg.burgstaller(-atsign-)i122server.vu-wien.ac.at

Journal and publication information

Publication Type: Journal Article; Research Support, Non-U.S. Gov't

Journal: BMC developmental biology (BMC Dev Biol), published in England. (Language: eng)

Reference: 2007-; vol 7 (issue ) : pp 141

Dates: Created 2008/03/07; Completed 2008/04/11; Revised 2008/11/20;

PMID: 18154666, status: MEDLINE (last retrieval date: 2/18/2009, IMS Date: )

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Animals Cloning, Organism - methods DNA, Mitochondrial - genetics Embryo, Mammalian Female Gene Dosage Inheritance Patterns

Microsatellite Repeats Nuclear Transfer Techniques Oocytes - cytology Polymerase Chain Reaction Pregnancy Sequence Analysis, DNA Sheep - embryology; genetics

Associated Chemicals: DNA, Mitochondrial (0)

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