Angiogenesis in the Regenerating Adult Zebrafish Tail Fin

Abstract

Angiogenesis is critical for regeneration and wound healing. Poor angiogenic responses to tissue injury lead to scarring or loss of tissue viability, which can be detrimental, sometimes even fatal in for example myocardial infarction or stroke. It is therefore of the outmost importance to find ways in which to speed the natural regenerative angiogenic response or improve it in patients exhibiting poor regeneration such as people suffering from diabetes. Zebrafish exhibit potent regenerative responses and are even able to regenerate tissues such as heart, brain and muscle tissue, which mammals cannot. Therefore, the zebrafish has recently become a popular model to study mechanisms behind regenerative angiogenesis and to find drugs that aid in or interfere with this process. Here we provide detailed protocols on how to study regenerative angiogenesis in the adult zebrafish tail fin. We give a detailed walk-through of the tail fin amputation itself as well as how to interfere with regeneration at the molecular level by injection of morpholinos or mRNA/expression vectors followed by electroporation. Finally we give suggestions on how to record the results and quantify the regeneration-induced angiogenesis. These protocols are quick and easy to use, and can be done with minimal training using equipment which is available in most zebrafish laboratories.