Salem Mohamed, Silverstein Jeff, Rexroad Caird E, Yao Jianbo
Laboratory of Animal Biotechnology and Genomics, Division of Animal and Nutritional Sciences, West Virginia University, Morgantown, WV 26505, USA.
BMC Genomics. 2007 Sep 19;8:328. doi: 10.1186/1471-2164-8-328.
Fast, efficiently growing animals have increased protein synthesis and/or reduced protein degradation relative to slow, inefficiently growing animals. Consequently, minimizing the energetic cost of protein turnover is a strategic goal for enhancing animal growth. Characterization of gene expression profiles associated with protein turnover would allow us to identify genes that could potentially be used as molecular biomarkers to select for germplasm with improved protein accretion.
We evaluated changes in hepatic global gene expression in response to 3-week starvation in rainbow trout (Oncorhynchus mykiss). Microarray analysis revealed a coordinated, down-regulated expression of protein biosynthesis genes in starved fish. In addition, the expression of genes involved in lipid metabolism/transport, aerobic respiration, blood functions and immune response were decreased in response to starvation. However, the microarray approach did not show a significant increase of gene expression in protein catabolic pathways. Further studies, using real-time PCR and enzyme activity assays, were performed to investigate the expression of genes involved in the major proteolytic pathways including calpains, the multi-catalytic proteasome and cathepsins. Starvation reduced mRNA expression of the calpain inhibitor, calpastatin long isoform (CAST-L), with a subsequent increase in the calpain catalytic activity. In addition, starvation caused a slight but significant increase in 20S proteasome activity without affecting mRNA levels of the proteasome genes. Neither the mRNA levels nor the activities of cathepsin D and L were affected by starvation.
These results suggest a significant role of calpain and 20S proteasome pathways in protein mobilization as a source of energy during fasting and a potential association of the CAST-L gene with fish protein accretion.
相对于生长缓慢、效率低下的动物,快速、高效生长的动物具有更高的蛋白质合成和/或更低的蛋白质降解率。因此,将蛋白质周转的能量成本降至最低是提高动物生长的一个战略目标。对与蛋白质周转相关的基因表达谱进行表征,将使我们能够识别那些有可能用作分子生物标志物的基因,以选择具有更高蛋白质沉积率的种质。
我们评估了虹鳟(Oncorhynchus mykiss)在3周饥饿后肝脏整体基因表达的变化。微阵列分析显示,饥饿鱼体内蛋白质生物合成基因的表达呈现出协同下调。此外,饥饿导致参与脂质代谢/运输、有氧呼吸、血液功能和免疫反应的基因表达下降。然而,微阵列方法并未显示蛋白质分解代谢途径中的基因表达有显著增加。我们进一步采用实时PCR和酶活性测定法进行研究,以探究参与主要蛋白水解途径的基因表达,这些途径包括钙蛋白酶、多催化蛋白酶体和组织蛋白酶。饥饿降低了钙蛋白酶抑制剂钙蛋白酶抑制蛋白长亚型(CAST-L)的mRNA表达,随后钙蛋白酶催化活性增加。此外,饥饿导致20S蛋白酶体活性略有但显著增加,而不影响蛋白酶体基因的mRNA水平。饥饿对组织蛋白酶D和L的mRNA水平及活性均无影响。
这些结果表明,钙蛋白酶和20S蛋白酶体途径在禁食期间作为能量来源的蛋白质动员中发挥重要作用,并且CAST-L基因与鱼类蛋白质沉积可能存在关联。