Abstract
To investigate the differentiation mechanism of grass carp preadipocytes, a primary adipocytes culture system was established. Confluent preadipocytes were induced to differentiation, and the morphology and gene expression were evaluated at different stages. It was shown that preadipocytes were gradually filled with droplets and the cellular lipid content increased during the differentiation. Ultrastructure observation indicated that the number of mitochondria increased with adipocytes differentiation. Consistently, the mitochondrial protein content was elevated in the differentiating adipocytes. qRT-PCR showed that the expression level of lipogenesis-related genes such as peroxisome proliferator activator receptor γ (PPAR γ), lipoprotein lipase (LPL), fatty acid synthase (FAS) and stearoyl-CoA desaturase (SCD) increased during adipocytes differentiation. The mitochondrial relevant gene also elevated when adipocyte differentiation, such as PPAR coactivator-1 (PGC-1 α), PGC-1 β and nuclear respiratory factor (NRF-1). However, the expression of carnitine palmitoyltransferase I α (CPT-1 α) gene decreased at the initial stage, but increased at the last stage of cell differentiation. These results indicated that the differentiation process of grass carp preadipocytes is similar to that of land animals, but the molecular mechanisms are not exactly the same. The findings revealed in this study provides new information to the study of fish adipocyte differentiation.
摘要
为了研究草鱼前体脂肪细胞的分化特征,分别收集不同分化阶段的脂肪细胞。油红O染色及提取法观测脂质蓄积情况;透射电镜观察细胞分化过程中胞内脂滴及线粒体的变化;提取细胞胞浆与线粒体蛋白分析线粒体蛋白含量的变化;实时定量PCR检测细胞脂质生成及线粒体发育相关基因的表达变化。结果显示,草鱼前体脂肪细胞在分化过程中,脂质含量逐渐且胞内脂滴增多,线粒体数量及蛋白含量也逐渐增加,脂质生成及线粒体发育相关基因表达水平也显著上调。说明草鱼前体脂肪细胞分化特征与哺乳动物相类似,细胞分化过程中线粒体的数量和蛋白含量增加,线粒体发育相关基因的表达水平也有所升高,表明线粒体在脂肪细胞分化过程中发挥着一定的作用。
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (31072223) and the National Basic Research Program of China (2014CB138603). The authors would thank Jingjing Tian, Caixia Lei, Wen Zhang, Xuexian Li for their assistance in the study.
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The authors declare that they have no conflict of interest.
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Liu, P., Ji, H., Li, C. et al. Morphology, mitochondrial development and adipogenic-related genes expression during adipocytes differentiation in grass carp (Ctenopharyngodon idellus). Sci. Bull. 60, 1241–1251 (2015). https://doi.org/10.1007/s11434-015-0833-9
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DOI: https://doi.org/10.1007/s11434-015-0833-9