The aquaculture of tilapia (Oreochromis sp.) is adversely affected by the sensitivity to cold stress. A large number of genes in tilapia were found to be regulated by cold stress, but their functions and mechanisms in cold tolerance remain largely unknown, partially due to the lack of a suitable in vitro model. An immortal neural cell line designated as TBN was established from brain tissue of the GIFT (Genetically Improved Farmed Tilapia) strain of Nile tilapia (Oreochromis niloticus). The TBN cells demonstrate a neuron-like morphology at low density and form a fibroblast-like monolayer at high density. Transcriptome profiling through RNA-sequencing revealed that a total of 15,011 genes were expressed in the TBN cells. The TBN cells express a wide array of marker genes for neural cells. A comparative analysis of the featured genes revealed the highest transcriptome similarity between the TBN cells and the TAPs (transient amplifying progenitors), among the 17 cell clusters isolated from the subventricular zone of mouse brain. The TBN cells tolerate relatively high culture temperatures and the highest growth rate was observed for the cells cultured at 32 o C in comparison with those at 30 o C, 28 o C and 26 o C. However, this cell line is cold sensitive. Exposure of the cells to 16 o C or lower temperatures significantly decreased cell confluences and induced apoptosis. The TBN cells were more sensitive to cold stress than the ZF4 cells (embryonic zebrafish fibroblasts). Moreover, the TBN cells can be efficiently transfected through electroporation. Our study provides an invaluable research tool to understand the nature of cold sensitivity of tilapia, and to dissect the function and mechanism of genes in regulating cold tolerance of fish. This article is protected by copyright. All rights reserved.

中文翻译：

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罗非鱼（Oreochromis niloticus）脑冷敏感神经细胞系的建立和表征

罗非鱼（Oreochromis sp.）的水产养殖受到冷应激敏感性的不利影响。罗非鱼中的大量基因被发现受冷胁迫调节，但它们在耐寒方面的功能和机制仍然很大程度上未知，部分原因是缺乏合适的体外模型。从尼罗罗非鱼（Oreochromis niloticus）的GIFT（遗传改良养殖罗非鱼）品系的脑组织中建立了一种称为TBN的永生神经细胞系。TBN 细胞在低密度下表现出神经元样形态，并在高密度下形成成纤维细胞样单层。通过 RNA 测序的转录组分析显示，TBN 细胞中总共表达了 15,011 个基因。TBN 细胞表达多种神经细胞标记基因。对特征基因的比较分析显示，在从小鼠脑室下区分离的 17 个细胞簇中，TBN 细胞和 TAP（瞬时放大祖细胞）之间的转录组相似性最高。TBN 细胞耐受相对较高的培养温度，与 30 o C、28 o C 和 26 o C 相比，32 o C 培养的细胞生长速率最高。然而，该细胞系对冷敏感。将细胞暴露于 16 o C 或更低的温度会显着降低细胞汇合并诱导细胞凋亡。TBN 细胞比 ZF4 细胞（胚胎斑马鱼成纤维细胞）对冷应激更敏感。此外，TBN 细胞可以通过电穿孔有效地转染。我们的研究为了解罗非鱼冷敏感性的性质，剖析基因在调节鱼类耐寒性中的功能和机制提供了宝贵的研究工具。本文受版权保护。版权所有。