1. Establishment of a heteroplasmic mouse strain with interspecific mitochondrial DNA haplotypes and improvement of a PCR-RFLP-based measurement system for estimation of mitochondrial DNA heteroplasmy
- Author
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Hiroshi Shitara, Hiromichi Yonekawa, Midori Yamaguchi, Liqin Cao, and Choji Taya
- Subjects
0301 basic medicine ,Mitochondrial DNA ,Non-Mendelian inheritance ,Microinjections ,Zygote ,Mitochondrion ,Biology ,DNA, Mitochondrial ,Polymerase Chain Reaction ,law.invention ,Mice ,03 medical and health sciences ,law ,Genetics ,Animals ,Heteroduplex formation ,Polymerase chain reaction ,Homoplasmy ,Molecular biology ,Heteroplasmy ,030104 developmental biology ,Haplotypes ,Female ,Animal Science and Zoology ,Restriction fragment length polymorphism ,Agronomy and Crop Science ,Polymorphism, Restriction Fragment Length ,Biotechnology - Abstract
Mitochondrial DNA segregation is one of the characteristic modes of mitochondrial inheritance in which the heteroplasmic state of mitochondrial DNA is transmitted to the next generation in variable proportions. To analyze mitochondrial DNA segregation, we produced a heteroplasmic mouse strain with interspecific mitochondrial DNA haplotypes, which contains two types of mitochondrial DNA derived from C57BL/6J and Mus spretus strains. The strain was produced on a C57BL/6J nuclear genomic background by microinjection of donor cytoplasm into fertilized eggs. The PCR-RFLP semi-quantitative analysis method, which was improved to reduce the effect of heteroduplex formation, was used to measure the proportion of heteroplasmic mitochondrial DNA in tissues. Founder mice contained up to approximately 14% of exogenous Mus spretus mitochondrial DNA molecules in their tails, and the detected proportions differed among tissues of the same individual. Heteroplasmic mitochondrial DNA is transmitted to the next generation in varying proportions under the maternal inheritance mode. This mitochondrial heteroplasmic mouse strain and the improved PCR-RFLP measurement system enable analysis of the transmission of heteroplasmic mitochondrial DNA variants between tissues and generations.
- Published
- 2017