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The Alternative Reading Frame (ARF) protein is a tumor suppressor encoded by the  Cyclin Dependent Kinase Inhibitor 2A  gene in mammals but not lower regenerative vertebrates, and has been previously implicated as a context-sensitive suppressor of regeneration in murine skeletal muscle and humanized ARF-expressing zebrafish fins. This study extends our investigation of the role of ARF in the regeneration of other solid tissues, including the zebrafish heart and the mammalian digit. Heart regeneration after cryoinjury was used to mimic massive myocardial infarction.  ARF  gene expression was upregulated during the cardiac regenerative process and slowed the rate of morphological recovery.  ARF  specifically impacts cardiomyocytes, neovascularization, and the endothelial-mesenchymal transition, while not affecting epicardial proliferation. This suggests that in the context of regeneration,  ARF  is specifically expressed in cells undergoing dedifferentiation. To investigate  ARF  as a suppressor of epimorphic regeneration in mammalian systems, we also tested whether the absence of  ARF  was permissive for murine digit regeneration, but found that  ARF  absence alone was insufficient to significantly alter digit restoration. These findings provide additional evidence that  ARF  suppresses epimorphic regeneration, but suggests that modulation of  ARF  alone is insufficient to permit regeneration.

Human ARF Specifically Inhibits Epimorphic Regeneration in the Zebrafish Heart