冠刺星（COTS; Acanthaster物种）是一种缓慢移动的腐蚀珊瑚生物，其长而锐利的毒棘提供了保护。毒素感染可能导致恶心、麻木、呕吐、关节疼痛，有时还会导致瘫痪。小分子皂苷和plancitoxin蛋白被认为与COTS的毒性有关。盐水虾致死率测定用于确认毒棘分泌的毒素生物分子。组织学分析结合基于毒棘的蛋白质组学有助于解释蛋白质的来源和身份，而定量RNA测序和系统发育学分别确认目标基因表达和相对保守性。我们展示了COTS毒棘分泌的生物分子对盐水虾的致死作用，包括使用>10 kDa（p>0.05, 9.82 µg/ml）的口部棘半纯化提取物的显著毒性，支持毒素蛋白质的分泌存在。COTS口部棘的超微结构观察表明存在能促进分泌蛋白质分布的孔隙。随后对棘源性蛋白质进行纯化和质谱分析发现了许多分泌蛋白质，包括plancitoxins以及在棘中基因表达相对较高的磷脂酶A2、蛋白酶抑制剂16样蛋白、ependymin相关蛋白和尚未被鉴定的蛋白质。一些分泌蛋白质（例如，卵黄蛋白和致命脑肿瘤蛋白1）在棘组织中的表达并不高，然而棘可能作为贮存或释放的场所。本研究通过对冠刺星口部棘的功能性、超微结构和蛋白质组学分析，为我们理解冠刺星做出了贡献。©2023 Hillberg等。

The crown-of-thorns starfish (COTS; Acanthaster species) is a slow-moving corallivore protected by an extensive array of long, sharp toxic spines. Envenomation can result in nausea, numbness, vomiting, joint aches and sometimes paralysis. Small molecule saponins and the plancitoxin proteins have been implicated in COTS toxicity.Brine shrimp lethality assays were used to confirm the secretion of spine toxin biomolecules. Histological analysis, followed by spine-derived proteomics helped to explain the source and identity of proteins, while quantitative RNA-sequencing and phylogeny confirmed target gene expression and relative conservation, respectively.We demonstrate the lethality of COTS spine secreted biomolecules on brine shrimp, including significant toxicity using aboral spine semi-purifications of >10 kDa (p > 0.05, 9.82 µg/ml), supporting the presence of secreted proteins as toxins. Ultrastructure observations of the COTS aboral spine showed the presence of pores that could facilitate the distribution of secreted proteins. Subsequent purification and mass spectrometry analysis of spine-derived proteins identified numerous secretory proteins, including plancitoxins, as well as those with relatively high gene expression in spines, including phospholipase A2, protease inhibitor 16-like protein, ependymin-related proteins and those uncharacterized. Some secretory proteins (e.g., vitellogenin and deleted in malignant brain tumor protein 1) were not highly expressed in spine tissue, yet the spine may serve as a storage or release site. This study contributes to our understanding of the COTS through functional, ultrastructural and proteomic analysis of aboral spines.©2023 Hillberg et al.