Understanding the mechanisms of neuronal survival in hypoxia-tolerant animals is crucial for developing effective treatments for ischemia-related pathologies. The freshwater fish Carassius gibelio exhibits remarkable resistance to hypoxia, making it an ideal model for studying neuronal calcium homeostasis under oxygen-deprived conditions. Disruptions in calcium signaling are implicated in hypoxia-induced neuronal apoptosis, with calcium removal mechanisms such as the Na+-Ca2+ exchanger (NCX), plasma membrane Ca2+-ATPase (PMCA), and mitochondrial sequestration playing critical roles in maintaining cytosolic calcium balance. In this study, we investigated the function of these calcium-regulating mechanisms in Carassius gibelio cerebellar neurons. Using microfluorescent calcium imaging and pharmacological inhibition, we found that both PMCA and NCX contribute significantly to cytoplasmic calcium clearance, with their inhibition resulting in a marked increase in basal intracellular calcium levels. Mitochondria also played a crucial role in calcium homeostasis, as blocking their function led to an increase in calcium transient amplitude and a prolonged return to baseline levels. These findings suggest that Carassius gibelio cerebellar neurons possess robust calcium regulatory systems that may contribute to their high resistance to hypoxic stress. Further studies are needed to explore the interplay between these mechanisms and their role in neuronal survival.