Department of Animal Science, University of Connecticut, Storrs, CT 06269, USA.
Proteomics and Metabolomics Facility, Center for Open Research Resources and Equipment, University of Connecticut, Storrs, CT 06269, USA.
J Anim Sci. 2022 Aug 1;100(8). doi: 10.1093/jas/skac061.
Poor maternal nutrition during gestation can result in reduced offspring muscle growth and altered muscle metabolism. We hypothesized that over- or restricted-nutrition during gestation would alter the longissimus dorsi muscle (LM) proteome of offspring. Pregnant ewes were fed 60% (restricted), 100% (control), or 140% (over) of National Research Council requirements for total digestible nutrients from day 30 of gestation until parturition. Fetal (RES, CON, OVER) LM were collected at days 90 and 135 of gestation, or from offspring within 24 h of birth. Sarcoplasmic proteins were isolated, trypsin digested, and subjected to multiplexed, label-based quantitative mass spectrometry analysis integrating tandem mass tag technology. Differential expression of proteins was identified by ANOVA followed by Tukey's HSD post hoc tests, and regularized regression via the elastic net. Significance was set at P < 0.05. Over-represented pathways containing differentially expressed proteins were identified by Reactome and included metabolism of proteins, immune system, cellular response to stress/external stimuli, developmental biology, and infectious disease. As a result of maternal diet, a total of 312 proteins were differentially expressed (day 90 = 89 proteins; day 135 = 115 proteins; birth = 131 proteins). Expression of eukaryotic initiation factor (EIF) 2S3, EIF3L, and EIF4G2 was lower in OVER fetuses at day 90 of gestation (P < 0.05). Calcineurin A and mitogen-activated protein kinase 1 were greater in RES fetuses at day 90 (P < 0.04). At day 135 of gestation, pyruvate kinase and lactate dehydrogenase A expression were greater in OVER fetuses than CON (P < 0.04). Thioredoxin expression was greater in RES fetuses relative to CON at day 135 (P = 0.05). At birth, proteins of the COP9 signalosome complex were greater in RES offspring relative to OVER (P < 0.05). Together, these data indicate that protein degradation and synthesis, metabolism, and oxidative stress are altered in a time and diet-specific manner, which may contribute to the phenotypic and metabolic changes observed during fetal development and postnatal growth.
妊娠期间母体营养不足可导致后代肌肉生长减少和肌肉代谢改变。我们假设妊娠期间过度或限制营养会改变后代背最长肌（LM）的蛋白质组。从妊娠第 30 天到分娩，怀孕的母羊分别接受 60%（限制）、100%（对照）或 140%（过度）的国家研究委员会规定的总可消化养分。在妊娠第 90 天和第 135 天或在分娩后 24 小时内从胎儿（RES、CON、OVER）LM 中收集肌肉。分离肌浆蛋白，胰蛋白酶消化，然后进行基于串联质量标签技术的多重、标记定量质谱分析。通过方差分析（ANOVA） followed by Tukey's HSD post hoc tests 和弹性网络（elastic net）进行差异蛋白的鉴定。显著水平设置为 P < 0.05。通过 Reactome 鉴定包含差异表达蛋白的高度代表性途径，包括蛋白质代谢、免疫系统、细胞对压力/外部刺激的反应、发育生物学和传染病。由于母体饮食，总共 312 种蛋白差异表达（第 90 天=89 种蛋白；第 135 天=115 种蛋白；出生时=131 种蛋白）。在妊娠第 90 天，OVER 胎儿的真核起始因子（EIF）2S3、EIF3L 和 EIF4G2 的表达较低（P < 0.05）。在第 90 天，RES 胎儿的钙调神经磷酸酶 A 和丝裂原活化蛋白激酶 1 较高（P < 0.04）。在妊娠第 135 天，OVER 胎儿的丙酮酸激酶和乳酸脱氢酶 A 的表达高于 CON（P < 0.04）。在妊娠第 135 天，相对于 CON，RES 胎儿的硫氧还蛋白表达较高（P=0.05）。在出生时，RES 后代的 COP9 信号体复合物蛋白较高（P < 0.05）。总的来说，这些数据表明，蛋白质降解和合成、代谢和氧化应激以时间和饮食特异性的方式发生改变，这可能导致在胎儿发育和产后生长过程中观察到的表型和代谢变化。
