Coexistence of Bos taurus and B. indicus Mitochondrial DNAs in Nuclear Transfer-Derived Somatic Cattle Clones

Corresponding author: Ralf Steinborn, University of Veterinary Medicine, Veterinärplatz 1, A-1210 Vienna, Austria., Ralf.Steinborn{at}vu-wien.ac.at (E-mail)

Communicating editor: J. VAN ARENDONK

We investigated the mitochondrial DNA (mtDNA) composition in one of the largest adult somatic mammalian clones (n = 20) reported so far. The healthy cloned cattle were derived from nuclear transfer of an identical nuclear genetic background (mural granulosa donor cells including surrounding cytoplasm) into enucleated oocytes with either Bos indicus or B. taurus mtDNA. Here we report the first cases of coexisting mtDNAs of two closely related subspecies following nuclear transfer. Heteroplasmy (0.6–2.8%) was found in 4 out of 11 cross-subspecies cloned cattle. Quantitation was performed using "amplification refractory mutation system (ARMS) allele-specific real-time PCR." We determined that the ratio of donor cell to recipient cytoplast mtDNA copy number was 0.9% before nuclear transfer. Therefore, we concluded that the percentage of donor cell mtDNA in the heteroplasmic intersubspecific cloned animals is in accordance with neutral transmission of donor mtDNA. We determined an amino acid sequence divergence of up to 1.3% for the two subspecies-specific mtDNA haplotypes. In addition, intrasubspecific B. indicus heteroplasmy of 1% (but up to 7.3 and 12.7% in muscle and follicular cells of one animal) was detected in 7 out of the 9 B. indicus intrasubspecific cloned cattle.

This article has been cited by other articles:

				A. Genasetti, M. L. Valentino, V. Carelli, D. Vigetti, M. Viola, E. G. Karousou, G. V. Melzi d'Eril, G. De Luca, A. Passi, and F. Pallotti Assessing Heteroplasmic Load in Leber's Hereditary Optic Neuropathy Mutation 3460G->A/MT-ND1 with A Real-Time PCR Quantitative Approach J. Mol. Diagn., September 1, 2007; 9(4): 538 - 545. [Abstract] [Full Text] [PDF]

				E. J. Bowles, J.-H. Lee, R. Alberio, R. E. I. Lloyd, D. Stekel, K. H. S. Campbell, and J. C. St. John Contrasting Effects of in Vitro Fertilization and Nuclear Transfer on the Expression of mtDNA Replication Factors Genetics, July 1, 2007; 176(3): 1511 - 1526. [Abstract] [Full Text] [PDF]

				G. F. Mastromonaco, L. A. Favetta, L. C. Smith, F. Filion, and W. A. King The Influence of Nuclear Content on Developmental Competence of Gaur x Cattle Hybrid In Vitro Fertilized and Somatic Cell Nuclear Transfer Embryos Biol Reprod, March 1, 2007; 76(3): 514 - 523. [Abstract] [Full Text] [PDF]

				E.C. Spikings, J. Alderson, and J.C.St. John Transmission of mitochondrial DNA following assisted reproduction and nuclear transfer Hum. Reprod. Update, July 1, 2006; 12(4): 401 - 415. [Abstract] [Full Text] [PDF]

				Y.-H. Sun, S.-P. Chen, Y.-P. Wang, W. Hu, and Z.-Y. Zhu Cytoplasmic Impact on Cross-Genus Cloned Fish Derived from Transgenic Common Carp (Cyprinus carpio) Nuclei and Goldfish (Carassius auratus) Enucleated Eggs Biol Reprod, March 1, 2005; 72(3): 510 - 515. [Abstract] [Full Text] [PDF]

				R. S.F. Lee, A. J. Peterson, M. J. Donnison, S. Ravelich, A. M. Ledgard, N. Li, J. E. Oliver, A. L. Miller, F. C. Tucker, B. Breier, et al. Cloned Cattle Fetuses with the Same Nuclear Genetics Are More Variable Than Contemporary Half-Siblings Resulting from Artificial Insemination and Exhibit Fetal and Placental Growth Deregulation Even in the First Trimester Biol Reprod, January 1, 2004; 70(1): 1 - 11. [Abstract] [Full Text] [PDF]

				M. C. Gomez, J. A. Jenkins, A. Giraldo, R. F. Harris, A. King, B. L. Dresser, and C. E. Pope Nuclear Transfer of Synchronized African Wild Cat Somatic Cells into Enucleated Domestic Cat Oocytes Biol Reprod, September 1, 2003; 69(3): 1032 - 1041. [Abstract] [Full Text] [PDF]