Isolation and proteomic study of fish liver lipid droplets

Yuwei Sun, Jian Heng, Feng Liu, Shuyan Zhang, Pingsheng Liu
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Abstract

Lipid droplets (LDs) are a neutral lipid storage organelle that is conserved in almost all species. Excessive storage of neutral lipids in LDs is directly associated with many metabolic syndromes. Zebrafish is a better model animal for the study of LD biology due to its transparent embryonic stage compared to other organisms. However, the study of LDs in fish has been difficult due to the lack of specific LD marker proteins and the limitation of purification technology. In this paper, the purification and proteomic analysis of liver LDs of fish including zebrafish and Carassius auratus were performed for the first time. 259 and 267 proteins were identified respectively. Besides most of the identified proteins were reported in previous LD proteomes of mammals, indicating the similarity between mammal and fish LDs. We also identified many unique proteins of liver LDs in fish that are involved in the regulation of LD dynamics. Through morphological and biochemical analysis, we found that the marker protein Plin2 of zebrafish LD was located on LDs in Huh7 cells. These results will facilitate further study of LDs in fish and liver metabolic diseases using fish as a model animal.
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鱼肝脂滴的分离与蛋白质组学研究
脂滴(ld)是一种中性的脂质储存细胞器,在几乎所有物种中都是保守的。中性脂在ld中的过度储存与许多代谢综合征直接相关。与其他生物相比,斑马鱼具有透明的胚胎阶段,是研究LD生物学的较好模型动物。然而,由于缺乏特异性的LD标记蛋白和纯化技术的限制,对鱼类LD的研究一直很困难。本文首次对斑马鱼、鲫鱼等鱼类的肝脏ld进行了纯化和蛋白质组学分析。分别鉴定出259个和267个蛋白。此外,所鉴定的蛋白质大部分是在哺乳动物LD蛋白质组中报道的,表明哺乳动物LD与鱼类LD具有相似性。我们还发现了许多鱼类肝脏LD的独特蛋白,这些蛋白参与了LD动力学的调节。通过形态学和生化分析,我们发现斑马鱼LD的标记蛋白Plin2位于Huh7细胞的LD上。这些结果将为进一步研究ld在鱼类中的作用以及以鱼类为模型动物的肝脏代谢疾病提供依据。
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