Time-lapse imaging of cell cycle dynamics during development in living cardiomyocyte

Hisayuki Hashimoto, Shinsuke Yuasa, Hidenori Tabata, Shugo Tohyama, Nozomi Hayashiji, Fumiyuki Hattori, Naoto Muraoka, Toru Egashira, Shinichiro Okata, Kojiro Yae, Tomohisa Seki, Takahiko Nishiyama, Kazunori Nakajima, Asako Sakaue-Sawano, Atsushi Miyawaki, Keiichi Fukuda

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Mammalian cardiomyocytes withdraw from the cell cycle shortly after birth, although it remains unclear how cardiomyocyte cell cycles behave during development. Compared to conventional immunohistochemistry in static observation, time-lapse imaging can reveal comprehensive data in hard-to-understand biological phenomenon. However, there are no reports of an established protocol of successful time-lapse imaging in mammalian heart. Thus, it is valuable to establish a time-lapse imaging system to enable the observation of cell cycle dynamics in living murine cardiomyocytes. This study sought to establish time-lapse imaging of murine heart to study cardiomyocyte cell cycle behavior. The Fucci (fluorescent ubiquitination-based cell cycle indicator) system can effectively label individual G1, S/G2/M, and G1/S-transition phase nuclei red, green and yellow, respectively, in living mammalian cells, and could therefore be useful to visualize the real-time cell cycle transitions in living murine heart. To establish a similar system for time-lapse imaging of murine heart, we first developed an ex vivo culture system, with the culture conditions determined in terms of sample state, serum concentration, and oxygen concentration. The optimal condition (slice culture, oxygen concentration 20%, serum concentration 10%) successfully mimicked physiological cardiomyocyte proliferation in vivo. Time-lapse imaging of cardiac slices from E11.5, E14.5, E18.5, and P1 Fucci-expressing transgenic mice revealed an elongated S/G2/M phase in cardiomyocytes during development. Our time-lapse imaging of murine heart revealed a gradual elongation of the S/G2/M phase during development in living cardiomyocytes.

Original languageEnglish
Pages (from-to)241-249
Number of pages9
JournalJournal of Molecular and Cellular Cardiology
Volume72
DOIs
Publication statusPublished - 2014 Jul

Keywords

  • Cardiac development
  • Cardiac regeneration
  • Cell culture
  • Cell cycle
  • Imaging

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

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    Hashimoto, H., Yuasa, S., Tabata, H., Tohyama, S., Hayashiji, N., Hattori, F., Muraoka, N., Egashira, T., Okata, S., Yae, K., Seki, T., Nishiyama, T., Nakajima, K., Sakaue-Sawano, A., Miyawaki, A., & Fukuda, K. (2014). Time-lapse imaging of cell cycle dynamics during development in living cardiomyocyte. Journal of Molecular and Cellular Cardiology, 72, 241-249. https://doi.org/10.1016/j.yjmcc.2014.03.020