Zebrafish are one of the creatures that have been increasingly considered in neurophysiological studies of pain as well as pharmacological and toxicological research. As the popularity of zebrafish grows in biomedical studies, the legal aspects of its applications have not grown at the same time and lagged behind. It seems that the evaluation of the legal aspects of Zebrafish applications in pharmaceutical and neuroscience research is important. The aim of this study was to investigate the ethical aspects of working with zebrafish in experiments such as the neurophysiology of pain and the effect of various drugs, and stress-related studies. Therefore, in this study, we will introduce the methods that have been considered in scientific research to reduce the pain and stress caused by experiments in zebrafish, including anesthesia and euthanasia techniques in this species.