Molting is a key step for body-size expansion and environmental adaptation of parasitic nematodes, and it is extremely important for Trichinella spiralis growth and development, but the molting mechanism is not fully understood. In this work, label-free LC-MS/MS was used to determine the proteome differences between T. spiralis muscle larvae (ML) at the encapsulated stage and intestinal infective larvae (IIL) at the molting stage. The results showed that a total of 2885 T. spiralis proteins were identified, 323 of which were differentially expressed. These proteins were involved in cuticle structural elements, regulation of cuticle synthesis, remodeling and degradation, and hormonal regulation of molting. These differential proteins were also involved in diverse intracellular pathways, such as fatty acid biosynthesis, arachidonic acid metabolism, and mucin type O-glycan biosynthesis. qPCR results showed that five T. spiralis genes (cuticle collagen 14, putative DOMON domain-containing protein, glutamine synthetase, cathepsin F and NADP-dependent isocitrate dehydrogenase) had significantly higher transcriptional levels in 10 h IIL than ML (P < 0.05), which were similar to their protein expression levels, suggesting that they might be T. spiralis molting-related genes. Identification and characterization of T. spiralis molting-related proteins will be helpful for developing vaccines and new drugs against the early enteral stage of T. spiralis.

中文翻译：

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旋毛虫肠道感染幼虫蜕皮相关蛋白的无标记定量蛋白质组学分析。

蜕皮是寄生线虫体大小扩展和环境适应的关键步骤，对于旋毛虫的生长和发育极为重要，但蜕皮机理尚未完全明了。在这项工作中，使用无标记的LC-MS / MS确定包裹阶段的螺旋旋毛肌幼虫（ML）和蜕皮阶段的肠感染性幼虫（IIL）之间的蛋白质组差异。结果表明，总共鉴定出2885个螺旋螺旋体蛋白，其中323个差异表达。这些蛋白质参与角质层的结构元素，角质层合成的调节，重塑和降解以及蜕皮的激素调节。这些差异蛋白还参与了多种细胞内途径，例如脂肪酸生物合成，花生四烯酸代谢，和粘蛋白型O-聚糖的生物合成。qPCR结果表明，五个螺旋藻基因（表皮胶原蛋白14，推定的含DOMON结构域蛋白，谷氨酰胺合成酶，组织蛋白酶F和NADP依赖性异柠檬酸脱氢酶）在10 h IIL中的转录水平显着高于ML（P <0.05），与它们的蛋白质表达水平相似，表明它们可能是螺旋旋毛虫蜕皮相关基因。螺旋藻蜕皮相关蛋白的鉴定和表征将有助于开发针对螺旋藻肠内早期阶段的疫苗和新药。组织蛋白酶F和NADP依赖性异柠檬酸脱氢酶在10 h IIL中的转录水平显着高于ML（P <0.05），这与其蛋白表达水平相似，表明它们可能与螺旋体蜕皮相关基因。螺旋藻蜕皮相关蛋白的鉴定和表征将有助于开发针对螺旋藻肠内早期阶段的疫苗和新药。组织蛋白酶F和NADP依赖性异柠檬酸脱氢酶在10 h IIL中的转录水平显着高于ML（P <0.05），这与其蛋白表达水平相似，表明它们可能与螺旋体蜕皮相关基因。螺旋藻蜕皮相关蛋白的鉴定和表征将有助于开发针对螺旋藻肠内早期阶段的疫苗和新药。