Regenerative growth in Eriocheir sinensis is driven by Slc7a5-TORC1-regulated cell signaling

Limb regeneration is a widespread phenomenon among animals, yet the underlying molecular mechanisms remain incompletely understood. Crustaceans like the Chinese mitten crab (Eriocheir sinensis), possess the remarkable ability to regrow lost limbs through a highly regulated regenerative process. In this study, we investigated the molecular mechanisms underlying limb regeneration in E. sinensis by integrating morphological and histological analyses, spatiotemporal mRNA expression profiling of TORC1 pathway genes, and RNA interference of Slc7a5 and TOR genes. Our results demonstrated that key genes within the TORC1 pathway, including Vatb, Rraga, Rragd, TOR, and S6K1, exhibit distinct expression patterns at different stages of limb regeneration, varying across developmental periods. Furthermore, in-situ hybridization and targeted knockdown of Slc7a5 and TOR genes significantly affected the rate of limb regeneration, highlighting their crucial roles in promoting blastema formation, papilla development, and ultimately, limb regeneration. Overall, our research reveals that Slc7a5 gene regulation within the TORC1 pathway significantly influences cell proliferation during limb regeneration in E. sinensis, thereby advancing our understanding of the role of TORC1 signaling in crustacean limb regeneration.