The pancreas plays important roles in the regulation of blood glucose, and is a well-studied organ in mammals because its dysfunction causes serious disorders, such as diabetes mellitus. However, mammals have the limited capacity for tissue regeneration in their organs, including pancreas. Fish may be an attractive model for regeneration studies, as fish exhibit a greater capacity for regeneration than do mammals. To elucidate the regenerative capacity of pancreatic β cells in medaka, we generated transgenic lines, in which β cells can be specifically ablated using the nitroreductase (NTR)/metronidazole (Mtz) system. We examined β-cell regeneration at embryonic-larval stages after specific ablation of β cells, and found that medaka rapidly regenerate β cells. Furthermore, we found that teleost-specific secondary islet have a unique feature in that their size increases in response to β-cell ablation in principal islets.