Osmoregulation of glutamine synthetase from Giant freshwater prawn (Macrobrachium rosenbergii) under osmotic stress

Zhijie, L.; Zhendong, Q.; Vijayaraman, S. B.; Chengkai, Y.; Guomao, S.; Jiabo, L.; Guang, Y.; Haiyang, S.; Gan, P.; Li, L.; Zhijie Lu, Zhendong Qin, Sarath Babu V, Chengkai Ye, Guomao Su, Jiabo Li, Guang Yang, Haiyang Shen,,

doi:10.1101/517409

bioRxiv · 10.1101/517409

Osmoregulation of glutamine synthetase from Giant freshwater prawn (Macrobrachium rosenbergii) under osmotic stress

Zhijie, L.·Zhendong, Q.·Vijayaraman, S. B.·Chengkai, Y.·Guomao, S.·Jiabo, L.·Guang, Y.·Haiyang, S.·Gan, P.·Li, L.·Zhijie Lu, Zhendong Qin, Sarath Babu V, Chengkai Ye, Guomao Su, Jiabo Li, Guang Yang, Haiyang Shen,,

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Abstract

Glutamine synthetase is a key enzyme that catalyzes the biosynthesis of glutamine (Gln) from glutamate and ammonia. Gln a vital amino acid acts as a precursor for protein synthesis and also assist in ammonia repressor and a key osmoregulators in aquatics. Here, we report the cloning and characterization of the GS gene from Macrobrachium rosenbergii (Mr-GS). The complete nucleotide and deduced amino acid sequences were determined that phylogenetically shared highest identity with other crustaceans. GS mRNA was differentially expressed in 6 different tissues, with high to low order as muscle > gills > heart > stomach > brain > haemolymph. Mr-GS expression and the glutamine concentrations were analyzed in the gills and muscle tissues of prawn under hyper/hypo-osmotic stress conditions. Under hyper-osmotic stress, the mRNA expression of Mr-GS was significantly increased in both gills and muscle at 3, 6 and 12 h post-treatment with 2.54, 4.21 and 10.83 folds, and 11.66, 17.97 and 45.92 folds, respectively. Protein analysis by western blot (WB) and Immunohistochemistry (IHC) further confirmed the Mr-GS expression was increased at 12 h post treatment. On the other hand, under hypo-osmotic stress, the mRNA expression of Mr-GS was also significantly increased in both gills and muscle at 3, 6 and 12 h post treatment with 1.63, 3.30 and 3.52 folds, and 4.06, 42.99 and 26.69 folds, respectively. Furthermore, under hyperosmotic stress, Gln concentration was increased in both gills and muscle at 6 and 12 h post treatment with 1.83, 2.02 folds, and 1.41, 1.29 folds, respectively. While, under hypo-osmotic stress, Gln concentration was increased in both gills and muscle at 3, 6 and 12 h post treatment with 3.99, 3.40, 2.59 folds, and 1.72, 1.83, 1.80 folds, respectively. Taken together, these results suggest that Mr-GS might play a key role in osmoregulation in M. rosenbergii.

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Zhijie, L., Zhendong, Q., Vijayaraman, S. B., Chengkai, Y., Guomao, S., Jiabo, L., Guang, Y., Haiyang, S., Gan, P., Li, L., Zhijie Lu, Zhendong Qin, Sarath Babu V, Chengkai Ye, Guomao Su, Jiabo Li, Guang Yang, Haiyang Shen,,. 2019-01-10. Osmoregulation of glutamine synthetase from Giant freshwater prawn (Macrobrachium rosenbergii) under osmotic stress. https://doi.org/10.1101/517409

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