Harvey Thomas N, Sandve Simen R, Jin Yang, Vik Jon Olav, Torgersen Jacob S
Centre for Integrative Genetics, Department of Animal and Aquacultural Sciences, Faculty of Biosciences, Norwegian University of Life Sciences, Ås, Norway.
AquaGen AS, Trondheim, Norway.
PeerJ. 2019 Sep 17;7:e7732. doi: 10.7717/peerj.7732. eCollection 2019.
Hepatic lipid metabolism is traditionally investigated in vitro using hepatocyte monocultures lacking the complex three-dimensional structure and interacting cell types essential liver function. Precision cut liver slice (PCLS) culture represents an alternative in vitro system, which benefits from retention of tissue architecture. Here, we present the first comprehensive evaluation of the PCLS method in fish (Atlantic salmon, Salmo salar L.) and validate it in the context of lipid metabolism using feeding trials, extensive transcriptomic data, and fatty acid measurements. We observe an initial period of post-slicing global transcriptome adjustment, which plateaued after 3 days in major metabolic pathways and stabilized through 9 days. PCLS fed alpha-linolenic acid (ALA) and insulin responded in a liver-like manner, increasing lipid biosynthesis gene expression. We identify interactions between insulin and ALA, where two PUFA biosynthesis genes that were induced by insulin or ALA alone, were highly down-regulated when insulin and ALA were combined. We also find that transcriptomic profiles of liver slices are exceedingly more similar to whole liver than hepatocyte monocultures, both for lipid metabolism and liver marker genes. PCLS culture opens new avenues for high throughput experimentation on the effect of "novel feed composition" and represent a promising new strategy for studying genotype-specific molecular features of metabolism.
传统上,肝脂质代谢是在体外使用缺乏复杂三维结构和对肝功能至关重要的相互作用细胞类型的肝细胞单培养物进行研究的。精密肝切片(PCLS)培养代表了一种替代的体外系统,它受益于组织结构的保留。在这里,我们首次对鱼类(大西洋鲑鱼,Salmo salar L.)的PCLS方法进行了全面评估,并通过喂养试验、广泛的转录组数据和脂肪酸测量在脂质代谢背景下对其进行了验证。我们观察到切片后全球转录组有一个初始调整期,在主要代谢途径中3天后趋于平稳,并在9天内保持稳定。用α-亚麻酸(ALA)和胰岛素喂养的PCLS以类似肝脏的方式做出反应,增加脂质生物合成基因的表达。我们确定了胰岛素和ALA之间的相互作用,其中两个单独由胰岛素或ALA诱导的多不饱和脂肪酸生物合成基因,在胰岛素和ALA联合使用时高度下调。我们还发现,无论是脂质代谢还是肝脏标记基因,肝切片的转录组谱都比肝细胞单培养物与全肝的转录组谱更为相似。PCLS培养为高通量研究“新型饲料成分”的效果开辟了新途径,并代表了一种研究代谢基因型特异性分子特征的有前途的新策略。