Establishment of conditional reporter mouse lines at ROSA26 locus for live cell imaging

Abe, Takaya; Kanazawa, Hiroshi; Shioi, Go; Inoue, Kazuaki; Nakao, Kazuki; Aizawa, Shinichi; Fujimori, Toshihiko

doi:10.1002/dvg.20753

Cited by 224 publications

(229 citation statements)

References 46 publications

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“…A reporter gene, coding for histone H2B fused to a mCherry fluorescent molecule, was introduced into the experimental mouse strain by backcrossing with reporter mice carrying a H2B-mCherry fusion gene 28 . Timelapse imaging of oocytes undergoing second meiotic division was performed in Ca 2 þ -free KSOM medium supplemented with 10 mM of SrCl 2 using a Leica DMI6000 microscope at 37°C, 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

Merotelic attachments allow alignment and stabilization of chromatids in meiosis II oocytes

2014

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The chromosome segregation process in human oocytes is highly error-prone, generating meiosis II (MII) oocytes with unbalanced chromatids that contribute to aneuploidy in offspring. This raises questions regarding the mechanism for transmission of chromatids and how chromatids evade the error correction mechanisms in MII oocytes. Here, we analyse the behaviour of chromatids in mouse MII oocytes. We find that chromatids align at the spindle equator at the metaphase stage of MII and that their presence does not obstruct entry into the anaphase stage. The alignment process is mediated by merotelic (bi-directional) microtubule-kinetochore attachments, revealing a multi-domain organization of the kinetochore of mammalian meiotic chromosomes. Our results suggest that biorientation of chromatids stabilize microtubule attachments at the kinetochores in a tension-dependent manner. Our results also suggest that merotelic attachments contribute to chromosome mis-segregation in wild-type MII oocytes. Thus, merotely is an important promoter of aneuploidy in mammalian oocytes.

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Section: Methodsmentioning

confidence: 99%

Merotelic attachments allow alignment and stabilization of chromatids in meiosis II oocytes

2014

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“…Combining our gene-trap lines with a transgenic mouse line ubiquitously expressing histone H2B tagged with mCherry (R26-H2B-mCherry [42]), we normalized the Venus signal to the mCherry signal, thereby correcting for systematic error arising from nuclear position within the embryo. Furthermore, to rightly assign a given cell position and fate [32], we used mTmG to obtain membrane signal (hereafter written as mT in this study [43]).…”

Section: Establishment Of the Lineage Segregation Map Of Mouse Pre-immentioning

confidence: 99%

Venus trap in the mouse embryo reveals distinct molecular dynamics underlying specification of first embryonic lineages

et al. 2015

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Mammalian development begins with the segregation of embryonic and extra-embryonic lineages in the blastocyst. Recent studies revealed cell-to-cell gene expression heterogeneity and dynamic cell rearrangements during mouse blastocyst formation. Thus, mechanistic understanding of lineage specification requires quantitative description of gene expression dynamics at a singlecell resolution in living embryos. However, only a few fluorescent gene expression reporter mice are available and quantitative live image analysis is limited so far. Here, we carried out a fluorescence gene-trap screen and established reporter mice expressing Venus specifically in the first lineages. Lineage tracking, quantitative gene expression and cell position analyses allowed us to build a comprehensive lineage map of mouse pre-implantation development. Our systematic analysis revealed that, contrary to the available models, the timing and mechanism of lineage specification may be distinct between the trophectoderm and the inner cell mass. While expression of our trophectoderm-specific lineage marker is upregulated in outside cells upon asymmetric divisions at 8-and 16-cell stages, the inside-specific upregulation of the inner-cell-mass marker only becomes evident at the 64-cell stage. This study thus provides a framework toward systems-level understanding of embryogenesis marked by high dynamicity and stochastic variability.

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“…We successfully established three Amh-Treck Tg lines (#36, #85, #94) with testis-specific expression of DTR in male pups (Supplementary Figure S1, see section on supplementary data given at the end of this article). The C57BL/6-Tg (CAG-EGFP) mice (GFP mice; SLC) and C57BL/6-R26-H2B-mCherry knockin mice (Cherry mice; Abe et al 2011) were used as transplant donors for the rescue experiment. The ICR and C57BL/6 male mice (E12.5-10 weeks old; SLC) were also used for the immunohistochemical analyses.…”

Section: Amh-treck Tg Mouse Linementioning

confidence: 99%

A novel Amh-Treck transgenic mouse line allows toxin-dependent loss of supporting cells in gonads

Shinomura¹,

Kishi²,

Tomita³

et al. 2014

Reproduction

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Cell ablation technology is useful for studying specific cell lineages in a developing organ in vivo. Herein, we established a novel anti-Mü llerian hormone (AMH)-toxin receptor-mediated cell knockout (Treck) mouse line, in which the diphtheria toxin (DT) receptor was specifically activated in Sertoli and granulosa cells in postnatal testes and ovaries respectively. In the postnatal testes of Amh-Treck transgenic (Tg) male mice, DT injection induced a specific loss of the Sertoli cells in a dose-dependent manner, as well as the specific degeneration of granulosa cells in the primary and secondary follicles caused by DT injection in Tg females. In the testes with depletion of Sertoli cell, germ cells appeared to survive for only several days after DT treatment and rapidly underwent cell degeneration, which led to the accumulation of a large amount of cell debris within the seminiferous tubules by day 10 after DT treatment. Transplantation of exogenous healthy Sertoli cells following DT treatment rescued the germ cell loss in the transplantation sites of the seminiferous epithelia, leading to a partial recovery of the spermatogenesis. These results provide not only in vivo evidence of the crucial role of Sertoli cells in the maintenance of germ cells, but also show that the Amh-Treck Tg line is a useful in vivo model of the function of the supporting cell lineage in developing mammalian gonads.

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Establishment of conditional reporter mouse lines at ROSA26 locus for live cell imaging

Cited by 224 publications

References 46 publications

Merotelic attachments allow alignment and stabilization of chromatids in meiosis II oocytes

Merotelic attachments allow alignment and stabilization of chromatids in meiosis II oocytes

Venus trap in the mouse embryo reveals distinct molecular dynamics underlying specification of first embryonic lineages

A novel Amh-Treck transgenic mouse line allows toxin-dependent loss of supporting cells in gonads

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