A constitutively expressed fluorescent ubiquitination-based cell-cycle indicator (FUCCI) in axolotls for studying tissue regeneration

Timothy J. Duerr, Eun Kyung Jeon, Kaylee M. Wells, Antonio Villanueva, Ashley W. Seifert, Catherine D. McCusker, James R. Monaghan

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Regulation of cell cycle progression is essential for cell proliferation during regeneration following injury. After appendage amputation, the axolotl (Ambystoma mexicanum) regenerates missing structures through an accumulation of proliferating cells known as the blastema. To study cell division during blastema growth, we generated a transgenic line of axolotls that ubiquitously expresses a bicistronic version of the fluorescent ubiquitination-based cell-cycle indicator (FUCCI). We demonstrate near-ubiquitous FUCCI expression in developing and adult tissues, and validate these expression patterns with DNA synthesis and mitosis phase markers. We demonstrate the utility of FUCCI for live and whole-mount imaging, showing the predominantly local contribution of cells during limb and tail regeneration. We also show that spinal cord amputation results in increased proliferation at least 5 mm from the site of injury. Finally, we use multimodal staining to provide cell type information for cycling cells by combining fluorescence in situ hybridization, EdU click-chemistry and immunohistochemistry on a single FUCCI tissue section. This new line of animals will be useful for studying cell cycle dynamics using in situ endpoint assays and in vivo imaging in developing and regenerating animals.

Original languageEnglish
JournalDevelopment (Cambridge)
Volume149
Issue number6
DOIs
StatePublished - Mar 15 2022

Bibliographical note

Publisher Copyright:
© 2022. Published by The Company of Biologists Ltd.

Keywords

  • Axolotl
  • Cell cycle
  • FUCCI
  • Regeneration

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology

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