Epithelial cell fusion is required for tissue repair following UV-A irradiation

Abstract

Cell cycle-dependent and independent mechanisms lead to the generation of mononucleated and multinucleated, polyploid cells. The more than doubling of a cells nuclear genome by endoreplication has been found to be an adaptation to genotoxic stress, enabling cell survival despite DNA damage. However, it remains unknown whether cells that increase ploidy via multinucleation also arise in response to genotoxic stress. Here, we use ultraviolet light A (UV-A) to induce permanent DNA damage in cells within the adult fruit fly epithelium. UV-A irradiation causes an injury-like response where giant multinucleated, polyploid cells arise following cell death. The epithelial cells undergo endoreplication, which is required to restore tissue mass, but is surprisingly dispensable for tissue repair. UV-A irradiation also induces cell fusion, which generates multinucleated cells that encompass almost the entire epithelial area post injury. Cell fusion can be inhibited by expression of a dominant negative Rac or Cdc42 GTPase, which then blocks epithelial tissue repair post irradiation. Apoptotic nuclei were detected at the site of cell junction breakdown suggesting that apoptosis itself or an apoptotic signal is required for polyploidization in this model. Expression of the effector caspase inhibitor, p35, led to inhibition of apoptosis, the endocycle, and cell fusion post UV-A. Therefore, we have discovered that caspase activation is necessary for polyploidization post injury and enhancing cell ploidy via multinucleation is another strategy to enable cell survival and tissue repair following genotoxic stress. Significance StatementPolyploid cells are lifes stress responders as the more than doubling of a cells genome has been shown to enable resistance to genotoxic stress. Cells are exposed to various sources of genotoxic stress, including from ultraviolet light. Here we find that ultraviolet light induces DNA damage causing apoptosis and the subsequent generation of giant, multinucleated polyploid cells in the fruit fly epithelium. Unlike in other models studied to date, cell fusion is the predominant response to genotoxic stress and appears to be essential for tissue repair. This study also determines that polyploidization post UV stress is dependent on caspase activation, suggesting a conserved mechanism to initiate cellular multinucleation both in development as well as in response to lifes stressors.