Establishment and characterization of a cold-sensitive neural cell line from the brain of tilapia (Oreochromis niloticus)

Abstract

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°C compared with those at 30°C, 28°C and 26°C. Nonetheless, this cell line is cold sensitive. Exposure of the cells to 16°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.

Keywords: cold sensitivity, neural cell line, optimal growth condition, tilapia, transcriptome, transfection efficiency.