title: 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.
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.
authors.affiliation: Laboratory for Transgenic Technology, Tokyo Metropolitan Institute of Medical Science, 1-6, Kamikitazawa 2-chome, Setagaya-ku, Tokyo, 156-8506, Japan. shitara-hr@igakuken.or.jp.|Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, 305-8572, Japan. shitara-hr@igakuken.or.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shitara
authors.affiliation: International Institute for Integrative Sleep Medicine (WPI-IIIS), University of Tsukuba, Tsukuba, Ibaraki, 305-8575, Japan.
authors.forename: Liqin
authors.initials: L
authors.lastname: Cao
authors.affiliation: Laboratory for Transgenic Technology, Tokyo Metropolitan Institute of Medical Science, 1-6, Kamikitazawa 2-chome, Setagaya-ku, Tokyo, 156-8506, Japan.
authors.forename: Midori
authors.initials: M
authors.lastname: Yamaguchi
authors.affiliation: Laboratory for Transgenic Technology, Tokyo Metropolitan Institute of Medical Science, 1-6, Kamikitazawa 2-chome, Setagaya-ku, Tokyo, 156-8506, Japan.
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Yonekawa
authors.affiliation: Laboratory for Transgenic Technology, Tokyo Metropolitan Institute of Medical Science, 1-6, Kamikitazawa 2-chome, Setagaya-ku, Tokyo, 156-8506, Japan.
authors.forename: Choji
authors.initials: C
authors.lastname: Taya
mesh_terms: D000818:Animals; D004272:DNA, Mitochondrial / Q000235:genetics*; D005260:Female; D006239:Haplotypes / Q000235:genetics*; D051379:Mice; D008845:Microinjections; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length / Q000235:genetics*; D015053:Zygote / Q000254:growth & development
chemical_list: D004272:DNA, Mitochondrial
keywords: Heteroplasmy; Mitochondria; Mitochondrial DNA; Mouse; PCR-RFLP