Humans, like all mammals, lack the natural ability to replace lost myocardium with new contractile tissue after cardiac damage. This deficiency contributes to heart failure, the primary cause of morbidity and mortality worldwide. However, recent studies suggest that the adult mammalian heart generates cardiomyocytes (CMs) throughout life, indicating an innate capacity for low levels of myocardial regeneration. Understanding the mechanisms of endogenous cardiac regeneration is fundamental to discovering better human therapeutic strategies. Zebrafish and neonatal mice represent excellent vertebrate model organisms for identifying the cellular and molecular mechanisms that underlie heart regeneration. They fully regenerate functional cardiac tissues without permanent scars after ventricular resection surgery, cryoinjury, or genetic ablation within several weeks. In the past 20 years, researchers have developed various critical zebrafish and neonatal mouse heart injury methods. This Special Issue of JoVE focusing on “Injury methods in cardiac regenerative models” will provide an essential platform to capture state-of-the-art techniques for heart regeneration studies. We are currently seeking protocols for heart injury methods, including but not limited to zebrafish, mice, and humans.