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Mitochondria transfer into mouse ova by microinjection

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Abstract

A method for mitochondria isolation and interspecific transfer of mitochondria was developed in mice. Mitochondria were isolated from Mus spretus liver samples for microinjection into fertilized ova obtained from superovulated M. musculus domesticus females. Electron microscopic observations of mitochondria preparations used for microinjection demonstrated intact mitochondrial vesicles with little microsomal contamination. Species-specific nested PCR primers complementary to sequence differences in the mitochondrial DNA D-loop region revealed high rates of successful transfer of foreign mitochondria after isolation and injection into zygotes cultured through the blastocyst stage of embryonic development. Of 217 zygotes, 67 survived mitochondria injection and 23 out of 37 zygotes developed were at the blastocyst-stage of embryonic development after 4.5 days of in vitro culture. All 23 of these blastocysts contained detectable levels of foreign mitochondria. These results represent an initial step in developing a model system to study mitochondrial dynamics and development of therapeutic strategies for human metabolic diseases affected by aberrations in mitochondrial function or mutation

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References

  • Berdanier, C.D. and Kim, M.J.C. (1993) Hyperthyroidism does not induce an increase in mitochondrial respiration in BHE/cdb rats. J. Nutr. Biochem. 4, 10–9.

    Google Scholar 

  • Brinster, R.L., Chen, H.Y., Trumbauer, M.E., Yagle, M.K. and Palmiter, R.D. (1985) Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs. Proc. Natl Acad. Sci. USA 82, 4438–42.

    Google Scholar 

  • Clayton, D.A. (1991) Replication and transcription of vertebrate mitochondrial DNA. Annu. Rev. Cell Biol. 7, 453–78.

    Google Scholar 

  • DeVivo, D.C. (1993) The expanding clinical spectrum of mitochondrial diseases. Brain Dev. 15, 1–22.

    Google Scholar 

  • Ebert, K.M., Alcivar, A., Liem, H. and Hecht, N.B. (1989) Mouse zygotes injected with mitochondria develop normally but the exogenous mitochondria are not detectable in the progeny. Molec. Reprod. Develop. 1, 156–63.

    Google Scholar 

  • Gyllensten, U., Wharton, D., Josefsson, A. and Wilson, A.C. (1991) Paternal inheritance of mitochondrial DNA in mice. Nature 352, 255–7.

    Google Scholar 

  • Hayashi, J.I., Ohta, S., Kikuchi, A., Takemitsu, M., Goto, Y.I. and Nonaka, I. (1991) Introduction of disease-related mitochondrial DNA deletions into HeLa cells lacking mitochondrial DNA results in mitochondrial dysfunction. Proc. Natl Acad. Sci. USA 88, 10614–8.

    Google Scholar 

  • Jenuth, J.P., Peterson, A.C., Fu, K. and Shoubridge, E.A. (1996) Random genetic drift in the female germline explains the rapid segregation of mammalian mitochondrial DNA. Nature Genet. 14, 146–51.

    Google Scholar 

  • Johnson, D. and Lardy, H. (1967) Isolation of liver and kidney mitochondria. Meth. Enzymol. 10, 94–6.

    Google Scholar 

  • Kaneda, H., Hayashi, J.I., Takahama, S., Taya, C., Lindahl, K.F. and Yonekawa, H. (1995) Elimination of paternal mitochondrial DNA in intraspecific crosses during early mouse embryogenesis. Proc. Natl Acad. Sci. USA 92, 4542–6.

    Google Scholar 

  • King, M.P. and Attardi, G. (1988) Injection of mitochondria into human cells leads to rapid replacement of the endogenous mitochondrial DNA. Cell 52, 811–9.

    Google Scholar 

  • Laipis, P.J. (1996) Construction of heteroplasmic mice containing two mitochondrial DNA genotypes by micromanipulation of single-cell embryos. Meth. Enzymol. 264, 345–57.

    Google Scholar 

  • Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–75.

    Google Scholar 

  • Meirelles, F.V. and Smith, I.C. (1997) Mitochondrial genotype segregation in a mouse heteroplasmic lineage produced by embryonic karyoplast transplantation. Genetics 145, 445–51.

    Google Scholar 

  • Palva, T.K. and Palva, E.T. (1985) Rapid isolation of animal mitochondrial DNA by alkaline extraction. FEBS Letters 192, 267–70.

    Google Scholar 

  • Polites, H.G. and Pinkert, C.A. (1994) DNA microinjection and transgenic animal production. In Pinkert, C.A. ed., Transgenic Animal Technology: a Laboratory Handbook, pp. 15–68. San Diego, CA: Academic Press.

    Google Scholar 

  • Shoffner, J.M. and Wallace, D.C. (1990) Oxidative phosphorylation diseases. Adv. Hum. Genetics 19, 267–330.

    Google Scholar 

  • Wallace, D.C. (1992) Diseases of the mitochondrial DNA. Annu. Rev. Biochem. 61, 1175–212.

    Google Scholar 

  • Yoneda, M., Miyatake, T. and Attardi, G. (1994) Complementation of mutant and wild-type human mitochondrial DNAs coexisting since the mutation event and lack of complementation of DNAs introduced separately into a cell with distinct organelles. Molec. Cell. Biol. 14, 2699–712.

    Google Scholar 

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PINKERT, C., IRWIN, M., JOHNSON, L. et al. Mitochondria transfer into mouse ova by microinjection. Transgenic Res 6, 379–383 (1997). https://doi.org/10.1023/A:1018431316831

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  • DOI: https://doi.org/10.1023/A:1018431316831

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