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Mosaic variegated aneuploidy in mouse BubR1 deficient embryos and pregnancy loss in human

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An Erratum to this article was published on 30 September 2014

Abstract

Chromosome aberrations (aneuploidies mostly) are the cause of the majority of spontaneous abortions in humans. However, little is known about defects in the underlying molecular mechanisms resulting in chromosome aberrations and following failure of preimplantation embryo development, initiation of implantation and postimplantation pregnancy loss. We suggest that defects of the spindle assembly checkpoint (SAC) are responsible for aneuploidy and the following abortions. To develop our hypothesis, we modeled this process in the mouse after inactivation of protein BubR1, one of the key players of SAC. We found that soon after implantation, more than 50 % of cells of BubR1 −/− embryos were aneuploid and had an increased level of premature sister chromatid separation (PSCS). Aneuploid cells do not have a predominant gain or loss of some specific chromosomes, but they have mosaic variegated aneuploidy (MVA), which is characterised by random mixture of different chromosomes. MVA leads to growth retardation, stochastic massive apoptosis, disruption of bilateral symmetry, and embryo death between embryonic days 7.5 to 13.5. Analysis published human data revealed that human recurrent pregnancy loss (RPL) embryos and rare infant patients carrying BubR1 mutations that have been described so far have the PSCS and MVA as in BubR1 deficient/insufficient mice. Based on this data, we predict that deficiency/insufficiency of BubR1 and other components of the SAC in human are responsible for a significant fraction of both early and late RPLs.

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Abbreviations

BubR1:

Bub1-related kinase

FISH:

Fluorescence in situ hybridization

IVF:

In vitro fertilization

MEFs:

Mouse embryonic fibroblasts

MVA:

Mosaic variegated aneuploidy

PSCS:

Premature sister chromatid separation

RPL:

Recurrent pregnancy loss

SAC:

Spindle assembly checkpoint

SKY:

Spectral karyotyping

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Acknowledgments

We gratefully acknowledge Winfried Edelmann, for providing advices. We thank Cornelia Leimeister, Barbara Zoeller and Galina Fedorova for expert technical assistance. We acknowledge Pamela Stanley for WW-6 ES cells. We are grateful to David Battaglia, Alexander Derkatch, Daniel Marks and Matt Thayer for critical reading of the manuscript and the helpful discussions. This work was partly supported, by the IZKF of the University of Wuerzburg, Germany and by the Office of the Vice President for Research, Oregon Health & Science University, Portland, OR, USA.

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Authors and Affiliations

  1. Department of Human Genetics, Biocenter, University of Würzburg, 97074, Würzburg, Germany

    Michael Schmid & Claus Steinlein

  2. Transgenic Mouse model Shared Resource, Knight Cancer Institute, Oregon Health & Science University, L481, 3181 SW Sam Jackson Park Road, Portland, OR, 97239, USA

    Qi Tian & Lev M. Fedorov

  3. Cytogenetics Research Laboratory, Department of Molecular and Medical Genetics, Oregon Health & Science University, Portland, OR, 97239, USA

    Amy E. Hanlon Newell & Susan B. Olson

  4. Developmental Biochemistry, Theodor-Boveri-Institute, Biocenter, University of Würzburg, 97074, Würzburg, Germany

    Manfred Gessler

  5. Institute of Pathology, University of Würzburg, 97080, Würzburg, Germany

    Andreas Rosenwald

  6. Physiological Chemistry, University of Würzburg, 97074, Würzburg, Germany

    Burkhard Kneitz & Lev M. Fedorov

  7. Department Urology and Pediatric Urology, University of Würzburg, 97080, Würzburg, Germany

    Burkhard Kneitz

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  1. Michael Schmid
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Fig. S1

Analysis of tumorigenesis in 18 month old BubR1 +/− mice. a,b Bronchioloalveolar lung adenoma in BubR1 +/− mouse. Small (a) and high (b) magnification, respectively. Hematoxylin-eosin staining. Scale bar: 420 μm (a), 60 μm (b). c Tumor incidence in different organs. (GIF 191 kb)

High resolution (TIFF 28654 kb)

Fig. S2

Karyotyping of BubR1 deficient embryos by G-banding. a, c, e Metaphases of E8.5 BubR1 −/− embryos after G- banding. b, d, f Karyograms of the aneuploid metaphases, 39, XY, -16; 42, XX, +6, +7,-16, +19, and euploid metaphase 40, XY, respectively. (GIF 114 kb)

High resolution (TIFF 3093 kb)

Fig. S3

Spectral karyotyping (SKY) of BubR1 knockout embryos. Metaphases with chromosome number 39 and 42 from Fig. 6 a-d are presented here in detail. (a-d) SKY of a BubR1 −/− embryo cell metaphase containing 39 chromosomes. a Metaphase after inverted DAPI staining. Y chromosomes are easily identified by their homogeneous dark staining (red arrows). b RGB image after hybridization with SKY-probes. c Pseudocolor image after per-pixel classification of the spectral data. d Karyogram of this metaphase showing RGB imaged (left), inverted DAPI stained (middle) and pseudo-colored chromosomes (right). Karyotyping indicates aneuploidy 39, XYY, -3,-10. (E-H) Spectral karypotyping of a BubR1 −/− embryo metaphase containing 42 chromosomes. e, f, g and h The images were produced as a, b, c, and d respectively. Karyotyping indicates aneuploidy; 42, XY, +8,+16. (GIF 288 kb)

High resolution (TIFF 21229 kb)

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Schmid, M., Steinlein, C., Tian, Q. et al. Mosaic variegated aneuploidy in mouse BubR1 deficient embryos and pregnancy loss in human. Chromosome Res 22, 375–392 (2014). https://doi.org/10.1007/s10577-014-9432-x

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