GeneCology Research Centre, School of Science and Engineering, University of the Sunshine Coast, 4 Locked Bag, Maroochydore, Queensland, 4558, Australia.
Institute for Marine and Antarctic Studies, University of Tasmania, Private Bag 49, Hobart, Tasmania, 7001, Australia.
Sci Rep. 2019 Feb 26;9(1):2847. doi: 10.1038/s41598-019-39173-7.
Literature on the cuticle formation in larval stages of the diverse group of decapod crustaceans is lacking, as opposed to a wealth of knowledge in several insect groups. Here we provide the first glimpse of the cuticular organisation in larvae of the eastern spiny lobster Sagmariasus verreauxi. A bioinformatic approach applied to S. verreauxi transcriptome through metamorphosis identified for the first time a small secreted protein with multiple isoforms that is highly expressed in crustacean larvae. This protein, named crustacean larval factor (Clf) shares structural characteristics with insect follicle cell protein 3 (FCP3), an insect-specific, rapidly evolving protein, with spatial-temporal regulated expression that is restricted to follicular cells during the production of the vitellin coat. Furthermore, we identified the FCP3 domain in additional structural proteins in multiple arthropod groups. Recombinant Clf inhibited in vitro calcium carbonate crystalline precipitation, in keeping with the finding that the spiny lobster larval cuticle is mainly composed of amorphous calcium carbonate. In addition, the recombinant Clf was shown to bind chitosan. Taken together, this research identifies two novel structural domains with lineage-specific expansion across arthropods. In crustaceans, Clf is found predominantly in larvae and the spatial-temporal regulated FCP3 factor occurs as a domain identified in multiple structural proteins across arthropods. Given the shared ten cysteines backbone between the Clf and FCP domains, a shared evolution is suggested and should be further explored.
关于十足目甲壳动物幼虫阶段的表皮形成的文献很少,而在几个昆虫群中却有大量的知识。在这里,我们首次观察到东方刺龙虾 Sagmariasus verreauxi 幼虫的表皮组织。通过变态对 S. verreauxi 转录组进行生物信息学分析,首次鉴定出一种小分泌蛋白,该蛋白具有多个同工型,在甲壳动物幼虫中高度表达。这种名为甲壳动物幼虫因子(Clf)的蛋白质与昆虫滤泡细胞蛋白 3(FCP3)具有结构特征,FCP3 是一种昆虫特异性、快速进化的蛋白质,其时空表达受到限制,仅在卵黄蛋白外壳生成期间存在于滤泡细胞中。此外,我们还在多个节肢动物群的其他结构蛋白中鉴定出了 FCP3 结构域。重组 Clf 抑制体外碳酸钙晶体沉淀,这与刺龙虾幼虫表皮主要由无定形碳酸钙组成的发现是一致的。此外,重组 Clf 被证明可以结合壳聚糖。总的来说,这项研究确定了两个具有谱系特异性扩张的新型结构域。在甲壳动物中,Clf 主要存在于幼虫中,而时空调节的 FCP3 因子则作为在多个节肢动物结构蛋白中鉴定出的结构域出现。鉴于 Clf 和 FCP 结构域之间共享的十个半胱氨酸骨架,提示它们具有共同的进化,并应进一步探讨。