Illuminating cell-cycle progression in the developing zebrafish embryo

Mayu Sugiyama, Asako Sakaue-Sawano, Tadahiro Iimura, Kiyoko Fukami, Tetsuya Kitaguchi, Koichi Kawakami, Hitoshi Okamoto, Shin Ichi Higashijima, Atsushi Miyawaki

Research output: Contribution to journalArticle

133 Citations (Scopus)

Abstract

By exploiting the cell-cycle-dependent proteolysis of two ubiquitination oscillators, human Cdt1 and geminin, which are the direct substrates of SCF Skp2 and APCCdh1 complexes, respectively, Fucci technique labels mammalian cell nuclei in G1 and S/G2/M phases with different colors. Transgenic mice expressing these G1 and S/G 2/M markers offer a powerful means to investigate the coordination of the cell cycle with morphogenetic processes. We attempted to introduce these markers into zebrafish embryos to take advantage of their favorable optical properties. However, although the fundamental mechanisms for cell-cycle control appear to be well conserved among species, the G1 marker based on the SCFSkp2-mediated degradation of human Cdt1 did not work in fish cells, probably because the marker was not ubiquitinated properly by a fish E3 ligase complex. We describe here the generation of a Fucci derivative using zebrafish homologs of Cdt1 and geminin, which provides sweeping views of cell proliferation in whole fish embryos. Remarkably, we discovered two anterior-to-posterior waves of cell-cycle transitions, G1/S and M/G1, in the differentiating notochord. Our study demonstrates the effectiveness of using the Cul4Ddb1-mediated Cdt1 degradation pathway common to all metazoans for the development of a G1 marker that works in the nonmammalian animal model.

Original languageEnglish
Pages (from-to)20812-20817
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number49
DOIs
Publication statusPublished - 2009 Dec 8
Externally publishedYes

Fingerprint

Geminin
Zebrafish
Cell Cycle
Fishes
Embryonic Structures
Notochord
Ubiquitin-Protein Ligases
G2 Phase
Ubiquitination
Cell Cycle Checkpoints
Cell Nucleus
Cell Division
Transgenic Mice
Proteolysis
Animal Models
Color
Cell Proliferation

Keywords

  • Cell cycle
  • Fluorescent protein
  • Imaging
  • Ubiquitination

ASJC Scopus subject areas

  • General

Cite this

Sugiyama, M., Sakaue-Sawano, A., Iimura, T., Fukami, K., Kitaguchi, T., Kawakami, K., ... Miyawaki, A. (2009). Illuminating cell-cycle progression in the developing zebrafish embryo. Proceedings of the National Academy of Sciences of the United States of America, 106(49), 20812-20817. https://doi.org/10.1073/pnas.0906464106

Illuminating cell-cycle progression in the developing zebrafish embryo. / Sugiyama, Mayu; Sakaue-Sawano, Asako; Iimura, Tadahiro; Fukami, Kiyoko; Kitaguchi, Tetsuya; Kawakami, Koichi; Okamoto, Hitoshi; Higashijima, Shin Ichi; Miyawaki, Atsushi.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 106, No. 49, 08.12.2009, p. 20812-20817.

Research output: Contribution to journalArticle

Sugiyama, M, Sakaue-Sawano, A, Iimura, T, Fukami, K, Kitaguchi, T, Kawakami, K, Okamoto, H, Higashijima, SI & Miyawaki, A 2009, 'Illuminating cell-cycle progression in the developing zebrafish embryo', Proceedings of the National Academy of Sciences of the United States of America, vol. 106, no. 49, pp. 20812-20817. https://doi.org/10.1073/pnas.0906464106
Sugiyama, Mayu ; Sakaue-Sawano, Asako ; Iimura, Tadahiro ; Fukami, Kiyoko ; Kitaguchi, Tetsuya ; Kawakami, Koichi ; Okamoto, Hitoshi ; Higashijima, Shin Ichi ; Miyawaki, Atsushi. / Illuminating cell-cycle progression in the developing zebrafish embryo. In: Proceedings of the National Academy of Sciences of the United States of America. 2009 ; Vol. 106, No. 49. pp. 20812-20817.
@article{39813d51d2cc43c2b1254adfdbdefe0d,
title = "Illuminating cell-cycle progression in the developing zebrafish embryo",
abstract = "By exploiting the cell-cycle-dependent proteolysis of two ubiquitination oscillators, human Cdt1 and geminin, which are the direct substrates of SCF Skp2 and APCCdh1 complexes, respectively, Fucci technique labels mammalian cell nuclei in G1 and S/G2/M phases with different colors. Transgenic mice expressing these G1 and S/G 2/M markers offer a powerful means to investigate the coordination of the cell cycle with morphogenetic processes. We attempted to introduce these markers into zebrafish embryos to take advantage of their favorable optical properties. However, although the fundamental mechanisms for cell-cycle control appear to be well conserved among species, the G1 marker based on the SCFSkp2-mediated degradation of human Cdt1 did not work in fish cells, probably because the marker was not ubiquitinated properly by a fish E3 ligase complex. We describe here the generation of a Fucci derivative using zebrafish homologs of Cdt1 and geminin, which provides sweeping views of cell proliferation in whole fish embryos. Remarkably, we discovered two anterior-to-posterior waves of cell-cycle transitions, G1/S and M/G1, in the differentiating notochord. Our study demonstrates the effectiveness of using the Cul4Ddb1-mediated Cdt1 degradation pathway common to all metazoans for the development of a G1 marker that works in the nonmammalian animal model.",
keywords = "Cell cycle, Fluorescent protein, Imaging, Ubiquitination",
author = "Mayu Sugiyama and Asako Sakaue-Sawano and Tadahiro Iimura and Kiyoko Fukami and Tetsuya Kitaguchi and Koichi Kawakami and Hitoshi Okamoto and Higashijima, {Shin Ichi} and Atsushi Miyawaki",
year = "2009",
month = "12",
day = "8",
doi = "10.1073/pnas.0906464106",
language = "English",
volume = "106",
pages = "20812--20817",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "49",

}

TY - JOUR

T1 - Illuminating cell-cycle progression in the developing zebrafish embryo

AU - Sugiyama, Mayu

AU - Sakaue-Sawano, Asako

AU - Iimura, Tadahiro

AU - Fukami, Kiyoko

AU - Kitaguchi, Tetsuya

AU - Kawakami, Koichi

AU - Okamoto, Hitoshi

AU - Higashijima, Shin Ichi

AU - Miyawaki, Atsushi

PY - 2009/12/8

Y1 - 2009/12/8

N2 - By exploiting the cell-cycle-dependent proteolysis of two ubiquitination oscillators, human Cdt1 and geminin, which are the direct substrates of SCF Skp2 and APCCdh1 complexes, respectively, Fucci technique labels mammalian cell nuclei in G1 and S/G2/M phases with different colors. Transgenic mice expressing these G1 and S/G 2/M markers offer a powerful means to investigate the coordination of the cell cycle with morphogenetic processes. We attempted to introduce these markers into zebrafish embryos to take advantage of their favorable optical properties. However, although the fundamental mechanisms for cell-cycle control appear to be well conserved among species, the G1 marker based on the SCFSkp2-mediated degradation of human Cdt1 did not work in fish cells, probably because the marker was not ubiquitinated properly by a fish E3 ligase complex. We describe here the generation of a Fucci derivative using zebrafish homologs of Cdt1 and geminin, which provides sweeping views of cell proliferation in whole fish embryos. Remarkably, we discovered two anterior-to-posterior waves of cell-cycle transitions, G1/S and M/G1, in the differentiating notochord. Our study demonstrates the effectiveness of using the Cul4Ddb1-mediated Cdt1 degradation pathway common to all metazoans for the development of a G1 marker that works in the nonmammalian animal model.

AB - By exploiting the cell-cycle-dependent proteolysis of two ubiquitination oscillators, human Cdt1 and geminin, which are the direct substrates of SCF Skp2 and APCCdh1 complexes, respectively, Fucci technique labels mammalian cell nuclei in G1 and S/G2/M phases with different colors. Transgenic mice expressing these G1 and S/G 2/M markers offer a powerful means to investigate the coordination of the cell cycle with morphogenetic processes. We attempted to introduce these markers into zebrafish embryos to take advantage of their favorable optical properties. However, although the fundamental mechanisms for cell-cycle control appear to be well conserved among species, the G1 marker based on the SCFSkp2-mediated degradation of human Cdt1 did not work in fish cells, probably because the marker was not ubiquitinated properly by a fish E3 ligase complex. We describe here the generation of a Fucci derivative using zebrafish homologs of Cdt1 and geminin, which provides sweeping views of cell proliferation in whole fish embryos. Remarkably, we discovered two anterior-to-posterior waves of cell-cycle transitions, G1/S and M/G1, in the differentiating notochord. Our study demonstrates the effectiveness of using the Cul4Ddb1-mediated Cdt1 degradation pathway common to all metazoans for the development of a G1 marker that works in the nonmammalian animal model.

KW - Cell cycle

KW - Fluorescent protein

KW - Imaging

KW - Ubiquitination

UR - http://www.scopus.com/inward/record.url?scp=73949083595&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=73949083595&partnerID=8YFLogxK

U2 - 10.1073/pnas.0906464106

DO - 10.1073/pnas.0906464106

M3 - Article

C2 - 19923430

AN - SCOPUS:73949083595

VL - 106

SP - 20812

EP - 20817

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 49

ER -