College of Life Science, Tianjin Normal University, Tianjin, 300387, China; Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal University, Tianjin, 300387, PR China.
College of Life Science, Tianjin Normal University, Tianjin, 300387, China.
Fish Shellfish Immunol. 2023 Sep;140:108981. doi: 10.1016/j.fsi.2023.108981. Epub 2023 Aug 4.
Tissue autotomy is a unique adaptive response to environmental stress, followed by regeneration process compensating for the loss of body parts. The crustaceans present remarkable activity of appendage autotomy and regeneration, however, the molecular mechanism is still unclear. In this study, the Eriocheir sinensis Hedgehog (EsHH) and Smoothened (EsSMO) were identified in the regenerative limbs, and the function of Hedgehog signaling pathway on limb regeneration was evaluated. At the blastema growth stage of limb regeneration, the expression of EsHH and EsSMO was up-regulated in response to limb autotomy stress, and down-regulated at blastema differentiation stage. To clarify the effect of Hedgehog pathway during limb regeneration, the regenerative efficiency was evaluated with Smoothened inhibitor cyclopamine or RNAi (ds-HH) injection. We observed that the regenerative efficiency was significantly repressed with blockage of Hedgehog pathway at both the basal growth stage and the proecdysial growth stage, which was indicated by the delay of wound healing and blastema growth, as well as a decrease in the size of newly formed limbs. In addition, gene expression and BrdU incorporation assay showed that the proliferation and myogenic differentiation of blastema cells were suppressed with either cyclopamine or ds-HH injection. Thus, these results suggest that Hedgehog signaling pathway is essential for the establishment of limb regeneration in E. sinensis through promoting the proliferation and myogenic differentiation of blastema cells.
组织自切是一种对环境压力的独特适应反应，随后是补偿身体部位损失的再生过程。甲壳动物的附肢自切和再生活性显著，但分子机制尚不清楚。在这项研究中，鉴定了中华绒螯蟹 Hedgehog（EsHH）和 Smoothened（EsSMO）在再生肢体中，并评估了 Hedgehog 信号通路对肢体再生的功能。在肢体再生的芽基生长阶段，EsHH 和 EsSMO 的表达在对肢体自切应激的反应中上调，并在芽基分化阶段下调。为了阐明 Hedgehog 通路在肢体再生过程中的作用，用 Smoothened 抑制剂环巴胺或 RNAi（ds-HH）注射评估了再生效率。我们观察到，Hedgehog 通路的阻断在基础生长阶段和蜕皮前生长阶段都显著抑制了再生效率，这表现为伤口愈合和芽基生长延迟，以及新形成肢体的大小减小。此外，基因表达和 BrdU 掺入测定表明，芽基细胞的增殖和肌源性分化均受到环巴胺或 ds-HH 注射的抑制。因此，这些结果表明 Hedgehog 信号通路通过促进芽基细胞的增殖和肌源性分化，对中华绒螯蟹的肢体再生建立是必不可少的。
