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在不同肌发生阶段的鱼类模型中对原代培养肌细胞进行蛋白质组学分析。

Proteomic characterization of primary cultured myocytes in a fish model at different myogenesis stages.

机构信息

Departament of Cell Biology, Physiology and Immunology, Faculty of Biology, University of Barcelona, 08028, Barcelona, Spain.

Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, 08028, Barcelona, Spain.

出版信息

Sci Rep. 2019 Oct 1;9(1):14126. doi: 10.1038/s41598-019-50651-w.

DOI:10.1038/s41598-019-50651-w
PMID:31576009
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6773717/
Abstract

Myogenesis is a complex two-phase process of proliferation and differentiation, which seems to be greatly conserved in vertebrates. For the first time in fish, we identify the changes that occur in the proteome during this process in a gilthead sea bream (Sparus aurata) myocyte primary cell culture (on days 4, 8 and 12), using 2-D gel electrophoresis and LC-MS/MS. A significant increase of myogenin expression at day 8 marked the transition from proliferation to differentiation. Of the 898 spots in the proteome analysis, the 25 protein spots overexpressed on day 4 and the 15 protein spots overexpressed on day 8 indicate the end of proliferation and the beginning of differentiation, respectively. Proliferation was characterized by enrichment of proteins involved in actin cytoskeleton remodelling and in cellular metabolic processes (transcription, ubiquitination, response to stress and glucose metabolism). During differentiation, 41 proteins were overexpressed and 51 underexpressed; many of them related to biosynthetic processes (RNA and protein synthesis and folding, and pentose pathways), terminal myotube formation and muscle contraction. The main cellular processes of both phases of muscle development in fish are similar with those observed in mammals but extended in time, allowing sequential studies of myogenesis.

摘要

肌发生是一个复杂的两阶段增殖和分化过程，在脊椎动物中似乎得到了很好的保守。我们首次在鱼类中海鲈鱼（Sparus aurata）肌原细胞原代培养（第 4、8 和 12 天）中，使用 2-D 凝胶电泳和 LC-MS/MS 鉴定了在这个过程中发生的蛋白质组变化。第 8 天肌生成素表达的显著增加标志着增殖向分化的转变。在蛋白质组分析的 898 个斑点中，第 4 天过度表达的 25 个蛋白质斑点和第 8 天过度表达的 15 个蛋白质斑点分别表示增殖的结束和分化的开始。增殖的特点是富含参与肌动蛋白细胞骨架重塑和细胞代谢过程（转录、泛素化、应激反应和葡萄糖代谢）的蛋白质。在分化过程中，有 41 种蛋白质过度表达，51 种蛋白质表达不足；其中许多与生物合成过程（RNA 和蛋白质合成和折叠，以及戊糖途径）、终末肌管形成和肌肉收缩有关。鱼类肌肉发育的两个阶段的主要细胞过程与在哺乳动物中观察到的过程相似，但时间延长，允许对肌发生进行连续研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/78d380f25a45/41598_2019_50651_Fig10_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/78d380f25a45/41598_2019_50651_Fig10_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/89c38512bdbd/41598_2019_50651_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/c1785433183c/41598_2019_50651_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/32c6576247fd/41598_2019_50651_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/1f01e76c2a48/41598_2019_50651_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/cf27f612de8f/41598_2019_50651_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/f1f5560a84ad/41598_2019_50651_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/78458edadf5b/41598_2019_50651_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/34267a9d23da/41598_2019_50651_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/6ed2fe866e0f/41598_2019_50651_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/89c3/6773717/78d380f25a45/41598_2019_50651_Fig10_HTML.jpg

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