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

Transgenic Research volume 26, pages559–565(2017)Cite this article

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.

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Acknowledgements

This work was supported by the Japan Society for the Promotion of Science KAKENHI Grant Numbers JP25430101 (to H.S.), JP25250011 (to H.Y.), and by the Takeda Science Foundation (to H.S.).

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Affiliations

  1. Laboratory for Transgenic Technology, Tokyo Metropolitan Institute of Medical Science, 1-6, Kamikitazawa 2-chome, Setagaya-ku, Tokyo, 156-8506, Japan

    Hiroshi Shitara, Midori Yamaguchi, Hiromichi Yonekawa & Choji Taya

  2. Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, 305-8572, Japan

    Hiroshi Shitara

  3. International Institute for Integrative Sleep Medicine (WPI-IIIS), University of Tsukuba, Tsukuba, Ibaraki, 305-8575, Japan

    Liqin Cao

Authors
  1. Hiroshi Shitara
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  2. Liqin Cao
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  3. Midori Yamaguchi
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  4. Hiromichi Yonekawa
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  5. Choji Taya
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Corresponding author

Correspondence to Hiroshi Shitara.

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Fig. S1

Supplementary Fig. S1 Proportion of B6-spr heteroplasmic mtDNA in tissues of F0 generation male mice (4-week-old). The male mice contained 6% of spr mtDNA in their tails at 3 weeks of age (Table 1). The histograms show the mean values from triplicates of each sample. #4 and #6 show individual numbers. In cerebrum, heart, and right soleus muscle of the #6 mouse; spr mtDNAs were not detected. (TIFF 64331 kb)

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Shitara, H., Cao, L., Yamaguchi, M. et al. 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. Transgenic Res 26, 559–565 (2017). https://doi.org/10.1007/s11248-017-0009-2

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Keywords

  • Mitochondrial DNA
  • Mitochondria
  • Heteroplasmy
  • PCR-RFLP
  • Mouse