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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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)).
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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.
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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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DOI: https://doi.org/10.1038/ng1970
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