Identification and characterisation of ROS modulator 1 in Lampetra japonica

Abstract

Reactive oxygen species (ROS) are a heterogeneous group of highly reactive molecules that oxidise targets in biological systems. ROS are also considered important immune regulators. In this study, we identified a homologue of reactive oxygen species modulator 1 (Romo1) in the Japanese lamprey (Lampetra japonica). The L japonica Romo1 (Lj-Romo1) gene shares high sequence homology with the Romo1 genes of jawed vertebrates. Real-time quantitative PCR demonstrated the wide distribution of Lj-Romo1 in lamprey tissues. Furthermore, after the lampreys were stimulated with lipopolysaccharide (LPS), the level of Lj-Romo1 mRNA was markedly up-regulated in the liver, gill, kidney, and intestine tissues. Lj-Romo1 was localised to the mitochondria and has the capacity to increase the ROS level in cells. The results obtained in the present study will help us to understand the roles of Romo1 in ROS production and innate immune responses in jawless vertebrates.

Keywords: 2′,7′- dichlorofluorescein; 2′,7′-dichlorofluorescein-diacetate; B-cell lymphoma-extra large; BLASTx; Basic Local Alignment Search Tool X; Bcl-X(L); DCF; DCFH-DA; ERK; ESTs; FBS; GAPDH; Innate immune; LPS; Lampetra japonica; Lampetra japonica Romo1; Lj-Romo1; MCF-7; ORF; PBS; PFA; ROS; ROS modulator 1; RPMI-1640; Reactive oxygen species; Romo1; Romo1 tagged with the enhanced green fluorescent protein; Romo1-EGFP; Roswell Park Memorial Institute-1640; TNF-α; expressed sequence tags; extracellular signal regulated kinase; fetal bovine serum; glyceraldehyde-3-phosphate dehydrogenase; human breast adenocarcinoma cell line; lipopolysaccharide; open reading frame; paraformaldehyde; phosphate-buffered saline; reactive oxygen species; tumor necrosis factor-alpha.