A novel murrel Channa striatus mitochondrial manganese superoxide dismutase: gene silencing, SOD activity, superoxide anion production and expression

Abstract

We have reported the molecular characterization including gene silencing, superoxide activity, superoxide anion production, gene expression and molecular characterization of a mitochondrial manganese superoxide dismutase (mMnSOD) from striped murrel Channa striatus (named as CsmMnSOD). The CsmMnSOD polypeptide contains 225 amino acids with a molecular weight of 25 kDa and a theoretical isoelectric point of 8.3. In the N-terminal region, CsmMnSOD carries a mitochondrial targeting sequence and a superoxide dismutases (SOD) Fe domain (28-109), and in C-terminal region, it carries another SOD Fe domain (114-220). The CsmMnSOD protein sequence shared significant similarity with its homolog of MnSOD from rock bream Oplegnathus fasciatus (96%). The phylogenetic analysis showed that the CsmMnSOD fell in the clade of fish mMnSOD group. The monomeric structure of CsmMnSOD possesses 9 α-helices (52.4%), 3 β-sheets (8.8%) and 38.8% random coils. The highest gene expression was noticed in liver, and its expression was inducted with fungal (Aphanomyces invadans) and bacterial (Aeromonas hydrophila) infections. The gene silencing results show that the fish that received dsRNA exhibited significant (P < 0.05) changes in expression when compared to their non-injected and fish physiological saline-injected controls. The SOD activity shows that the activity increases with the spread of infection and decreases once the molecule controls the pathogen. The capacity of superoxide anion production was determined by calculating the granular blood cell count during infection in murrel. It shows that the infection influenced the superoxide radical production which plays a major role in killing the pathogens. Overall, this study indicated the defense potentiality of CsmMnSOD; however, further research is necessary to explore its capability at protein level.

Wnt10b Participates in Regulating Fatty Acid Synthesis in the Muscle of Zebrafish

“…The double-stranded RNA (dsRNA) was synthesized with the primers Wnt10b-F3 and Wnt10b-R3 according to the method of Arockiaraj et al [12] (Table 1). Forty-eight male zebrafish were randomly distributed into eight glass tanks.…”

Investigation of the Lipid-Lowering Effect of Vitamin C Through GSK-3β/β-Catenin Signaling in Zebrafish

“…Firstly, the open reading frame (ORF) of GSK-3β gene (NM_131381.1) was cloned with the primers (F1: 5′-CTG GTG AGC AGT AGG GTG-3′; R1: 5′-CGG ATT CGT TCA AGA CAA). According to the method of Arockiaraj et al (2014) , the double-stranded RNA (dsRNA) was synthesized with the primers containing the T7 promoter RNA polymerase recognition sequence at the 5′ end (F2: 5′-GAT CAC TAA TAC GAC TCA CTA TAG GGC GGC ATT CGG CAG CAT GAA AG-3′; R2: 5′-GAT CAC TAA TAC GAC TCA CTA TAG GGG CAC GGC TGT GTC TGG GTC CA-3′). The reactions of in vitro transcription were performed in water at 37°C in a solution containing T7 RNA polymerase (Takara), 100 mM DTT, RNase Inhibitor, 5 × transcription buffer, 2.5 mM rNTP, and the template.…”

Innate and adaptive immune molecules of striped murrel Channa striatus

“…It revealed that the knockdown of MnSOD transcripts is dose‐dependent (Arockiaraj et al . ; Kumaresan et al . ).…”

“…Gene expression of CsMnSOD was analysed through gene silencing studies using interference RNA (RNAi) approach. It revealed that the knockdown of MnSOD transcripts is dose-dependent (Arockiaraj et al 2014d;Kumaresan et al 2015c).…”