NMDAR-CaMKII Pathway as a Central Regulator of Aggressiveness: Evidence from Transcriptomic and Metabolomic Analysis in Swimming Crabs Portunus trituberculatus.

Aggressiveness is one of the personality traits of crustaceans, playing a crucial role in their growth, life history, and adaptability by influencing resource acquisition. However, the neuroregulatory mechanisms of aggressiveness in crustaceans remain poorly understood. The thoracic ganglion offers valuable insights into complementary aspects of aggression control. This study identified the aggressiveness of swimming crabs Portunus trituberculatus, conducted transcriptomic and metabolomic analyses of the thoracic ganglia, and confirmed the neural regulatory effects on aggressiveness. Behavioral analyses showed that highly aggressive individuals exhibited increased frequency and duration of chela extension, more frequent attacks, approaches and retreats, as well as extended movement distances. Omics analysis revealed 11 key candidate genes and three metabolites associated with aggressiveness, which were primarily enriched in pathways related to energy metabolism and neurodegeneration. Injection of an NMDAR activator significantly decreased aggressiveness in highly aggressive crabs, accompanied by a significant increase in NMDAR protein fluorescence intensity and downregulation of NR2B, CaMKII, and CREB genes. Conversely, when lowly aggressive crabs were injected with an NMDAR inhibitor, they showed increased aggressiveness alongside significantly decreased NMDAR protein fluorescence intensity, upregulated NR2B expression, and downregulated CaMKII and CREB genes. These results suggest that NMDAR within the thoracic ganglia serves as a key receptor in modulating aggressiveness in P. trituberculatus, potentially by influencing neural energy state via the NMDAR-CaMKII pathway, which in turn affects oxidative phosphorylation, cAMP, and FoxO pathways.