The mitochondrial bottleneck occurs without reduction of mtDNA content in female mouse germ cells

Nature Genetics volume 39, pages386–390(2007)Cite this article

Abstract

Observations of rapid shifts in mitochondrial DNA (mtDNA) variants between generations prompted the creation of the bottleneck theory. A prevalent hypothesis is that a massive reduction in mtDNA content during early oogenesis leads to the bottleneck1,2. To test this, we estimated the mtDNA copy number in single germline cells and in single somatic cells of early embryos in mice. Primordial germ cells (PGCs) show consistent, moderate mtDNA copy numbers across developmental stages, whereas primary oocytes demonstrate substantial mtDNA expansion during early oocyte maturation. Some somatic cells possess a very low mtDNA copy number. We also demonstrated that PGCs have more than 100 mitochondria per cell. We conclude that the mitochondrial bottleneck is not due to a drastic decline in mtDNA copy number in early oogenesis but rather to a small effective number of segregation units for mtDNA in mouse germ cells. These results provide new information for mtDNA segregation models and for understanding the recurrence risks for mtDNA diseases.

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Figure 1: Models for rapid mtDNA segregation resulting from low segregation unit number in mouse female germ line.
Figure 2: Distribution of mtDNA copy number in single small primordial germ cells of mice from 7.5 to 13.5 dpc.
Figure 3: Distribution of mtDNA copy number in single cells of 5.5- to 6.5-dpc mouse embryos and in single somatic cells of 7.5- to 13.5-dpc mouse embryos.

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Acknowledgements

We thank N. Takahata and K. Fischer Lindahl for valuable comments and discussions. This work was supported in part by Grants-in-Aid (numbers 14GS0305 and 16770009) from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and national funds from the Ministry of Economy, Trade and Industry of Japan and the New Energy and Industrial Technology Development Organization (NEDO (the Nakano research group)).

Author information

Author notes

  1. Liqin Cao and Hiroshi Shitara: These authors contributed equally to this work.

Affiliations

  1. Department of Laboratory Animal Science, Tokyo Metropolitan Institute of Medical Science, Tokyo, 113-8613, Japan

    Liqin Cao, Hiroshi Shitara & Hiromichi Yonekawa

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

    Liqin Cao & Jun-Ichi Hayashi

  3. Laboratory of Reproductive Biology, Graduate School of Agriculture, Kyoto University, Kyoto, 606-8502, Japan

    Takuro Horii, Yasumitsu Nagao & Hiroshi Imai

  4. Technology and Development Team for Mammalian Cellular Dynamics, BioResource Center (BRC), RIKEN Tsukuba Institute, Ibaraki, 305-0074, Japan

    Kuniya Abe

  5. Stem Cell Project Group, Tokyo Metropolitan Institute of Medical Science, Tokyo, 113-8613, Japan

    Takahiko Hara

Authors
  1. Liqin Cao
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  2. Hiroshi Shitara
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  3. Takuro Horii
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  4. Yasumitsu Nagao
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  7. Takahiko Hara
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  8. Jun-Ichi Hayashi
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  9. Hiromichi Yonekawa
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Contributions

This study was designed by H.S., L.C., J.-I.H. and H.Y. and written by L.C., H.S. and H.Y. The transgenic mice were generated by K.A.; PGCs were prepared by H.S., L.C., T. Horii, Y.N., H.I. and T. Hara and other experimental procedures were done by L.C. and H.S.

Corresponding author

Correspondence to Hiromichi Yonekawa.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Distribution of mtDNA copy number in single M primordial germ cells from 7.5- to 13.5-dpc mice. (PDF 493 kb)

Supplementary Fig. 2

Distribution of mtDNA copy number in single L primordial germ cells from 7.5- to 13.5-dpc mice. (PDF 479 kb)

Supplementary Fig. 3

Mitochondria in 9.5-dpc mouse primordial germ cell. (PDF 1524 kb)

Supplementary Fig. 4

Mitochondria in 12.5-dpc mouse primordial germ cell. (PDF 2028 kb)

Supplementary Table 1

Primer and probe sequences. (PDF 12 kb)

Supplementary Video 1

Mitochondria in 9.5-dpc mouse primordial germ cell (lower cell). (MOV 895 kb)

Supplementary Video 2

Mitochondria in 12.5-dpc mouse primordial germ cell. (MOV 543 kb)

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Cao, L., Shitara, H., Horii, T. et al. The mitochondrial bottleneck occurs without reduction of mtDNA content in female mouse germ cells. Nat Genet 39, 386–390 (2007). https://doi.org/10.1038/ng1970

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