Exhaustive exploration of novel factors related to cardiomyocyte proliferation using the cell cycle-sensitive protein FUCCI

Abstract

【Background and Objective】

Since mammalian cardiomyocytes exit from cell cycle immediately after birth, the regenerative activity is limited in adult mammalian hearts. The aim of this study is to conduct a comprehensive search for pro-proliferative factors in mammalian cardiomyocytes, using a Fluorescent Ubiquitination-based Cell Cycle Indicator (FUCCI) that labels S/G2/M phases nuclei in green.

【Methods and Results】

Neonatal rat heart-derived cells were infected with the adenovirus vector expressing FUCCI or GFP. Immunofluorescence microscopic analysis showed that more than 95% of FUCCI or GFP positive cells represented cardiomyocytes. Next, FUCCI or GFP positive cells were obtained using FACS and analyzed by RNA-seq. One hundred and eight genes, including pro-proliferative factors such as various cyclins, were increased in FUCCI positive cells compared with GFP positive cells. Interestingly, we found that the expression of several genes related with cancer cell proliferation was also upregulated in FUCCI positive cells.

【Conclusion】

A system to comprehensively search for cell proliferation-related factors in cardiomyocytes was constructed using FUCCI expressing adenovirus and FACS.