INRAE, UR1037 LPGP, Fish Physiology and Genomics, Campus de Beaulieu, 35000, Rennes, France.
Université de Rennes 1, Campus de Beaulieu, 35000, Rennes, France.
Sci Rep. 2023 Jun 20;13(1):9967. doi: 10.1038/s41598-023-36354-3.
Xenopus egg extract is a powerful material to modify cultured cells fate and to induce cellular reprogramming in mammals. In this study, the response of goldfish fin cells to in vitro exposure to Xenopus egg extract, and subsequent culture, was studied using a cDNA microarray approach, gene ontology and KEGG pathways analyses, and qPCR validation. We observed that several actors of the TGFβ and Wnt/β-catenin signaling pathways, as well as some mesenchymal markers, were inhibited in treated cells, while several epithelial markers were upregulated. This was associated with morphological changes of the cells in culture, suggesting that egg extract drove cultured fin cells towards a mesenchymal-epithelial transition. This indicates that Xenopus egg extract treatment relieved some barriers of somatic reprogramming in fish cells. However, the lack of re-expression of pou2 and nanog pluripotency markers, the absence of DNA methylation remodeling of their promoter region, and the strong decrease in de novo lipid biosynthesis metabolism, indicate that reprogramming was only partial. The observed changes may render these treated cells more suitable for studies on in vivo reprogramming after somatic cell nuclear transfer.
爪蟾卵提取物是一种强大的物质,可以改变培养细胞的命运,并在哺乳动物中诱导细胞重编程。在这项研究中,我们使用 cDNA 微阵列方法、基因本体论和 KEGG 途径分析以及 qPCR 验证,研究了金鱼鳍细胞对体外暴露于爪蟾卵提取物以及随后的培养的反应。我们观察到 TGFβ 和 Wnt/β-catenin 信号通路的几个因子以及一些间充质标记物在处理的细胞中被抑制,而一些上皮标记物被上调。这与细胞在培养中的形态变化有关,表明卵提取物促使培养的鳍细胞向间充质-上皮转化。这表明爪蟾卵提取物处理减轻了鱼类细胞体细胞重编程的一些障碍。然而, pou2 和 nanog 多能性标记物的重新表达缺失、其启动子区域的 DNA 甲基化重塑缺失以及从头脂质生物合成代谢的强烈减少表明重编程仅部分发生。观察到的变化可能使这些处理过的细胞更适合用于核移植后体内重编程的研究。