Establishment and characterization of a cold-sensitive neural cell line from the brain of tilapia (Oreochromis niloticus)
Establishment and characterization of a cold-sensitive neural cell line from the brain of tilapia (Oreochromis niloticus)
复制标题
罗非鱼（Oreochromis niloticus）大脑冷敏感神经细胞系的建立和表征
DOI：
10.1111/jfb.14637
复制
发表时间：
2020-12-16
影响因子：
2
通讯作者：
Cui, Zongbin
中科院分区：
文献类型：
--
作者：
Long, Yong;Liu, Ran;Cui, Zongbin
罗非鱼（Oreochromis sp.）对冷胁迫的敏感性对其养殖产生不利影响。在罗非鱼中发现了大量受冷胁迫调控的基因，但它们在耐寒性中的功能和机制仍不清楚，部分原因是缺乏合适的体外模型。以尼罗罗非鱼（Oreochromis niloticus）转基因养殖罗非鱼品系脑组织为材料，建立了罗非鱼脑神经（TBN）永生神经细胞系。TBN细胞低密度呈神经元样形态，高密度呈成纤维细胞样单层。通过rna测序的转录组分析显示，TBN细胞中共有15011个基因表达。TBN细胞表达一系列神经细胞的标记基因。对从小鼠脑室下区分离的17个细胞簇的特征基因进行比较分析，发现TBN细胞与瞬时扩增祖细胞（TAPs）之间的转录组相似性最高。TBN细胞耐受较高的培养温度，与30℃、28℃和26℃培养的细胞相比，32℃培养的细胞生长速率最高，但该细胞系对冷敏感。将细胞暴露在16℃或更低的温度下显著降低细胞融合并诱导细胞凋亡。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，通过电穿孔可以有效地转染TBN细胞。本研究为了解罗非鱼冷敏感性的本质，揭示基因调控鱼类耐冷性的功能和机制提供了宝贵的研究工具。
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 tilapia brain neural (TBN) was established from brain tissue of the genetically improved farmed tilapia strain of Nile tilapia (Oreochromis niloticus). The TBN cells show 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 among the 17 cell clusters isolated from the subventricular zone of mouse brain revealed the highest transcriptome similarity between the TBN cells and the transient amplifying progenitors (TAPs). The TBN cells tolerate relatively high culture temperatures, and the highest growth rate was observed for the cells cultured at 32 degrees C compared with those at 30 degrees C, 28 degrees C and 26 degrees C. Nonetheless, this cell line is cold sensitive. Exposure of the cells to 16 degrees 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. This 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.