Investigating the Cellular and Molecular Mechanisms of Wound Healing in Oocytes and Embryos.

The African clawed frog has remarkable capacities to heal wounds rapidly and to regenerate complex tissues. Because of its experimental tractability, studies using oocytes, embryos, and larvae have contributed extensively to our understanding of the molecular and cellular mechanisms underpinning wound healing and tissue regeneration. In this protocol, we describe wound-healing assays following mechanical or laser injuries of oocytes and multicellular epithelia in embryos. We also explain how to perform assays aimed at investigating the cellular and molecular events during wound healing, including gene knockdown and overexpression experiments. In the latter assays, we explore the use of biochemical pull-down assays to investigate the activity of Rho GTPases, as well as the injection of mRNAs encoding fluorescent proteins or probes, followed by quantitative confocal image analyses to assay the dynamics of cytoskeletal components and their regulators.