Wei Fulei, Liang Jian, Tian Wengen, Yu Luxian, Feng Zhaohui, Hua Qiang
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, People's Republic of China.
State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University, 251 Ningda Road, Xining, 810016, People's Republic of China.
Bioresour Bioprocess. 2022 Sep 26;9(1):104. doi: 10.1186/s40643-022-00589-1.
Gymnocypris przewalskii is the only high-land endemic teleost living in Qinghai Lake, the largest saline-alkaline lake in China. Its osmoregulatory physiology remains elusive due to a lack of precise identification of the response proteins. In the present study, DIA/SWATH was used to identify differentially expressed proteins (DEPs) under alkaline (pH = 10.1, carbonate buffer), saline (12‰, sodium chloride), and saline-alkaline [carbonate buffer (pH = 10.1) plus 11‰ sodium chloride] stresses. A total of 66,056 unique peptides representing 7,150 proteins and 230 DEPs [the false discovery rate (FDR) ≤ 0.05, fold change (FC) ≥ 1.5] were identified under different stresses. Comparative analyses of the proteome and transcriptome indicated that over 86% of DEPs did not show consistent trends with mRNA. In addition to consistent enrichment results under different stresses, the specific DEPs involved in saline-alkaline adaptation were primarily enriched in functions of homeostasis, hormone synthesis and reactions of defense response, complement activation and reproductive development. Meanwhile, a protein-protein interaction (PPI) network analysis of these specific DEPs indicated that the hub genes were ITGAX, MMP9, C3, F2, CD74, BTK, ANXA1, NCKAP1L, and CASP8. This study accurately isolated the genes that respond to stress, and the results could be helpful for understanding the physiological regulation mechanisms regarding salinity, alkalinity, and salinity-alkalinity interactions.
普氏裸鲤是中国最大的盐碱湖——青海湖中唯一的高原特有硬骨鱼类。由于缺乏对应答蛋白的精确鉴定，其渗透调节生理机制仍不清楚。在本研究中，采用数据独立采集/同步累积连续碎裂（DIA/SWATH）技术鉴定在碱性（pH = 10.1，碳酸盐缓冲液）、盐性（12‰，氯化钠）和盐碱（碳酸盐缓冲液（pH = 10.1）加11‰氯化钠）胁迫下的差异表达蛋白（DEP）。在不同胁迫下共鉴定出代表7150种蛋白质的66056条独特肽段和230个DEP（错误发现率（FDR）≤0.05，变化倍数（FC）≥1.5）。蛋白质组和转录组的比较分析表明，超过86%的DEP与mRNA没有一致的趋势。除了在不同胁迫下有一致的富集结果外，参与盐碱适应的特定DEP主要富集在稳态、激素合成以及防御反应、补体激活和生殖发育反应等功能中。同时，对这些特定DEP的蛋白质-蛋白质相互作用（PPI）网络分析表明，枢纽基因是整合素αX（ITGAX）、基质金属蛋白酶9（MMP9）、补体C3（C3）、凝血因子II（F2）、CD74分子（CD74）、布鲁顿酪氨酸激酶（BTK）、膜联蛋白A1（ANXA1）、NCK相关蛋白1样蛋白（NCKAP1L）和半胱天冬酶8（CASP8）。本研究准确分离出了应激反应基因，其结果有助于理解关于盐度、碱度以及盐碱相互作用的生理调节机制。
