The Potential Role of As-sumo-1 in the Embryonic Diapause Process and Early Embryo Development of Artemia sinica

Abstract

During embryonic development of Artemia sinica, environmental stresses induce the embryo diapause phenomenon, required to resist apoptosis and regulate cell cycle activity. The small ubiquitin-related modifier-1 (SUMO), a reversible post-translational protein modifier, plays an important role in embryo development. SUMO regulates multiple cellular processes, including development and other biological processes. The molecular mechanism of diapause, diapause termination and the role of As-sumo-1 in this processes and in early embryo development of Artemia sinica still remains unknown. In this study, the complete cDNA sequences of the sumo-1 homolog, sumo ligase homolog, caspase-1 homolog and cyclin B homolog from Artemia sinica were cloned. The mRNA expression patterns of As-sumo-1, sumo ligase, caspase-1, cyclin B and the location of As-sumo-1 were investigated. SUMO-1, p53, Mdm2, Caspase-1, Cyclin B and Cyclin E proteins were analyzed during different developmental stages of the embryo of A. sinica. Small interfering RNA (siRNA) was used to verify the function of sumo-1 in A. sinica. The full-length cDNA of As-sumo-1 was 476 bp, encoding a 92 amino acid protein. The As-caspases-1 cDNA was 966 bp, encoding a 245 amino-acid protein. The As-sumo ligase cDNA was 1556 bp encoding, a 343 amino acid protein, and the cyclin B cDNA was 739 bp, encoding a 133 amino acid protein. The expressions of As-sumo-1, As-caspase-1 and As-cyclin B were highest at the 10 h stage of embryonic development, and As-sumo ligase showed its highest expression at 0 h. The expression of As-SUMO-1 showed no tissue or organ specificity. Western blotting showed high expression of As-SUMO-1, p53, Mdm2, Caspase-1, Cyclin B and Cyclin E at the 10 h stage. The siRNA caused abnormal development of the embryo, with increased malformation and mortality. As-SUMO-1 is a crucial regulation and modification protein resumption of embryonic diapause and early embryo development of A. sinica.

The Potential Roles of the Apoptosis-Related Protein PDRG1 in Diapause Embryo Restarting of Artemia sinica

“…These results are consistent with the hypothesized function for As -PDRG1. The qPCR result agreed with the results of two other studies concerning genes relevant to the cell cycle in A. sinica [ 21 , 22 ].…”

RNA interference in shrimp and potential applications in aquaculture

“…; Hayakijkosol & Owens ; Chu et al . ). The chemical synthesis method allows siRNA production with a high quality and purity, and allows incorporating modifications to the backbone and/or RNA bases to improve siRNA target specificity and stability in the presence of nucleases.…”

“…Most likely, however, the resulting dsRNA will not be as pure as when synthesized in vitro as small amounts of bacterial remnants might always remain. Finally, siRNA oligos can also be chemically synthesized Hayakijkosol & Owens 2012;Chu et al 2014). The chemical synthesis method allows siRNA production with a high quality and purity, and allows incorporating modifications to the backbone and/or RNA bases to improve siRNA target specificity and stability in the presence of nucleases.…”

“…Embryos of A. sinica were electroporated in buffer containing siRNA, which is designed for targeting As-sumo-1 from A. sinica. Resulting deformations appeared at the nauplius stage, and the deformed individuals all continued to survive (Chu et al 2014). Electroporation allows the delivery of nucleic acid into embryos at an early stage and can be performed with large numbers of zygotes or embryos at the same time.…”

“…Artemia species have a short life cycle and survive in extreme conditions. RNAi studies on Artemia gradually unravel mechanisms of stress resistance and diapause cysts (Liu et al 2009;Robbins et al 2010;Dai et al 2011;King & MacRae 2012;Li et al 2012;Yang et al 2012;Zhao et al 2012;Zhou et al 2014), controlling the process of cell division (Dai et al 2008;Li et al 2012;Zhou et al 2013), genes involved in the development and differentiation (Copf et al 2004(Copf et al , 2006Chu et al 2014) and reproductive mode (Dai et al 2010).…”