Pt-LIS1 participates nuclear deformation and acrosome formation via regulating Dynein-1 during spermatogenesis in Portunus trituberculatus

Spermatogenesis involves complex dynamic mechanisms. Dynein-1 is a key carrier in cellular cargo transport, participating in nuclear deformation and acrosome formation during spermatogenesis. However, the regulatory mechanisms of Dynein-1 during cargo transport remain unknown. In this study, we explored the role of Lissencephaly 1 (LIS1) in spermatogenesis and its impact on Dynein-1 cargo transport in Portunus trituberculatus. LIS1 was known as a Dynein-1 regulator, which is a causative gene of anencephaly syndrome. Pt-Lis1 was cloned from the crab testis, and its highly expression was observed in the testis. Pt-LIS1 dynamically localized around the nucleus and acrosome during spermatogenesis, colocalizing with Dynein-1 subunits, microtubules, mitochondrial markers (PHB), and Acrosin. RNA interference reduced Pt-Lis1 expression, leading to decreased expression of Pt-dhc and Pt-dic. During spermatogenesis, the signals of Pt-LIS1, Pt-DHC, Pt-DIC, and α-Tubulin were weakened and showed disorganized distribution. The colocalization of Pt-LIS1 with Pt-DHC and Pt-DIC decreased, while abnormal colocalization significantly increased. In addition, caspase-3 and p53 expression significantly increased after Pt-Lis1 silencing, indicating association with apoptosis in spermatogenic cells. All these results suggest that LIS1 played a crucial role in crustacean spermatogenesis by regulating nuclear deformation and acrosome formation through modulating Dynein-1 transport cargoes along microtubules.