Jiang Sufei, Qiao Hui, Fu Hongtuo, Gu Zemao
College of Fisheries, Shuangshui Shuanglü Institute, Huazhong Agricultural University, Wuhan 430070, China.
Hubei Hongshan Laboratory, Wuhan 430070, China.
Animals (Basel). 2023 Mar 8;13(6):977. doi: 10.3390/ani13060977.
A TMT-based (Tandem Mass Tag) liquid chromatography-tandem mass spectrometry (LC-MS/MS) proteomics approach was employed to explore differentially expressed proteins (DEPs) and KEGG pathways in hepatopancreas of 5 ovary stages. In total, 17,999 peptides were detected, among which 3395 proteins were identified. Further analysis revealed 26, 24, 37, and 308 DEPs in HE-I versus HE-II, HE-II versus HE-Ⅲ, HE-Ⅲ versus HE-Ⅳ, and HE-Ⅳ versus HE-Ⅴ, respectively (HE-I, HE-II, HE-III, HE-IV, and HE-V means hepatopancreas sampled from ovary stage I to V.). Gene ontology (GO) analysis indicated that DEPs were significantly enriched in "catalytic activity", "metabolic process", and "cell" of 4 comparison groups in turn. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment results showed that in hepatopancreas, as the ovaries developed to maturation, carbohydrate metabolism, lipid metabolism, amino acid metabolism, and lysosome played important roles in turn. The mRNA expression of 15 selected DEPs were consistent with proteome results by qPCR analysis. Further mRNA expression investigation results suggested 4 proteins (fatty acid-binding protein, NPC intracellular cholesterol transporter 1, Serine hydroxymethyltransferase, and Crustapin) were involved in ovary maturation. These results enhance the understanding of the regulatory role of hepatopancreas in ovary maturation and provide new insights for understanding the crustacean regulation mechanisms.
采用基于串联质量标签(TMT)的液相色谱-串联质谱(LC-MS/MS)蛋白质组学方法,探究5个卵巢发育阶段肝胰腺中差异表达蛋白(DEP)及京都基因与基因组百科全书(KEGG)通路。共检测到17999个肽段,鉴定出3395种蛋白质。进一步分析显示,在HE-I与HE-II、HE-II与HE-Ⅲ、HE-Ⅲ与HE-Ⅳ、HE-Ⅳ与HE-Ⅴ比较中,分别有26、24、37和308个差异表达蛋白(HE-I、HE-II、HE-III、HE-IV和HE-V分别表示从卵巢发育阶段I到V采集的肝胰腺)。基因本体(GO)分析表明,4个比较组的差异表达蛋白依次在“催化活性”“代谢过程”和“细胞”中显著富集。KEGG富集结果显示,在肝胰腺中,随着卵巢发育成熟,碳水化合物代谢、脂质代谢、氨基酸代谢和溶酶体依次发挥重要作用。通过qPCR分析,15个选定差异表达蛋白的mRNA表达与蛋白质组结果一致。进一步的mRNA表达研究结果表明,4种蛋白质(脂肪酸结合蛋白、NPC细胞内胆固醇转运蛋白1、丝氨酸羟甲基转移酶和甲壳素)参与卵巢成熟。这些结果加深了对肝胰腺在卵巢成熟中调控作用的理解,并为理解甲壳类动物调控机制提供了新的见解。