First cleavage plane of the mouse egg is not predetermined but defined by the topology of the two apposing pronuclei

Abstract

Studies of experimentally manipulated embryos1,2,3,4 have led to the long-held conclusion that the polarity of the mouse embryo remains undetermined until the blastocyst stage. However, recent studies5,6,7 reporting that the embryonic–abembryonic axis of the blastocyst arises perpendicular to the first cleavage plane, and hence to the animal–vegetal axis of the zygote, have led to the claim that the axis of the mouse embryo is already specified in the egg. Here we show that there is no specification of the axis in the egg. Time-lapse recordings show that the second polar body does not mark a stationary animal pole, but instead, in half of the embryos, moves towards a first cleavage plane. The first cleavage plane coincides with the plane defined by the two apposing pronuclei once they have moved to the centre of the egg. Pronuclear transfer experiments confirm that the first cleavage plane is not determined in early interphase but rather is specified by the newly formed topology of the two pronuclei. The microtubule networks that allow mixing of parental chromosomes before dividing into two may be involved in these processes.

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Figure 1: The 2pb moves towards the first cleavage plane in half the embryos.
Figure 2: The first cleavage plane is specified by the topology of the two apposing pronuclei in the egg centre.
Figure 3: Relationship between cytoskeleton, chromosomes and first cleavage plane specification.
Figure 4: First cleavage plane is not determined in early interphase but is specified according to the newly formed topology of the pronuclei after manipulation.

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Acknowledgements

We thank Y. Kaneda for HVJ; Z. Polanski, N. Bobola, A. Tomilin, P. Nielsen, R. Cassada and M. Hoffman for discussions and reading of the manuscript.

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Correspondence to Takashi Hiiragi.

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Supplementary information

Supplementary Movie 1

This movie shows a time-lapse recording of the embryos developing from zygote to 2-cell. The time-lapse corresponds to the sequential pictures of Fig.1b, covering a period of 7.5 hours. (MOV 1582 kb)

Supplementary Movie 2

This movie shows a time-lapse recording of the embryos developing from zygote to 2-cell. The time-lapse corresponds to the sequential pictures of Fig.1c, covering a period of 7.5 hours. (MOV 1155 kb)

Supplementary Movie 3

This movie shows a time-lapse recording of the embryo developing from zygote to 2-cell. The time-lapse corresponds to the sequential pictures of Fig.2a, covering a period of 17 hours. (MOV 2650 kb)

Supplementary Movie 4

This movie shows a time-lapse recording of the embryos developing from zygote to 2-cell. The time-lapse corresponds to the sequential pictures of Fig.2b, covering a period of 18 hours. (MOV 3305 kb)

Supplementary Movie 5

This movie shows a time-lapse recording of the embryos developing from zygote to 2-cell. The time-lapse corresponds to the sequential pictures of Fig.2c, covering a period of 19 hours. (MOV 4512 kb)

Supplementary Movie 6

This movie shows a time-lapse recording of the embryos developing from zygote to 2-cell following transfer of female pronucleus into fertilized egg from which male pronucleus was removed. Female pronucleus was placed in the pole opposite from 2pb. The time-lapse corresponds to the sequential pictures of Fig.4e, covering a period of 10 hours. (MOV 1967 kb)

Supplementary Methods

Describes the methods for pronuclear transfer and immunofluorescence staining. (DOC 20 kb)

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Hiiragi, T., Solter, D. First cleavage plane of the mouse egg is not predetermined but defined by the topology of the two apposing pronuclei. Nature 430, 360–364 (2004). https://doi.org/10.1038/nature02595

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