Determining the function of LvSmad3 on Litopenaeus vannamei in response to acute low temperature stress.

Smad3 is a key mediator of the canonical TGF-β signaling pathway and plays an important role in TGF-β1-mediated transcriptional regulation. However, the function of Smad3 in crustaceans such as shrimp, is still poorly understood and needs to be further explored. We characterized Litopenaeus vannamei Smad3 (LvSmad3) and its biological functions were investigated in response low temperature stress. Full-length LvSmad3 cDNA was 2341bp and contained an open reading frame (ORF) of 1326 bp that encoded a 441 amino acid long protein, with a predicted molecular mass of 48.35 kDa. Phylogenetic analysis revealed that LvSmad3 has a high degree of similarity with other known species. LvSmad3 mRNA was detected in all the tested tissues and highest transcription occurred mostly in gills. Further research showed that suppressing the expression of Smad3 could reduce ROS production, DNA damage and the apoptosis rate in shrimp hemocyte under low temperature compared with the dsGFP group. Thus, we speculated that Smad3 could promote the apoptosis of hemocytes. We confirmed that Smad3 could inhibit apoptosis in the hepatopancreas by suppressing the expression of pro-apoptotic genes. Taken together, the silencing of Smad3 can reduce ROS production induced by low temperature stress, weaken the damage to hemocytes and the hepatopancreas by inhibit the apoptosis.