Ultraviolet B retards growth, induces oxidative stress, and modulates DNA repair-related gene and heat shock protein gene expression in the monogonont rotifer, Brachionus sp.

Abstract

Ultraviolet B (UV-B) radiation causes direct cellular damage by breakage of DNA strands and oxidative stress induction in aquatic organisms. To understand the effect of UV-B radiation on the rotifer, Brachionus sp., several parameters including 24-h survival rate, population growth rate, and ROS level were measured after exposure to a wide range of UV-B doses. To check the expression of other important inducible genes such as replication protein A ( RPA), DNA-dependent protein kinase ( DNA-PK), Ku70, Ku80, and heat shock proteins ( hsps) after UV-B radiation, we observed dose- and time-dependency at 2 kJ/m 2. We also examined 13 hsp genes for their roles in the UV-B damaged rotifer. Results showed that UV-B remarkably inhibited the population growth of Brachionus sp. The level of intracellular reactive oxygen species (ROS) was high at 2 kJ/m 2, suggesting that 2 kJ/m 2 would already be toxic. This result was supported by other enzymatic activities, such as GSH levels, glutathione peroxidase, glutathione S-transferase, and glutathione reductase. For dose dependency, low doses of UV-B radiation (2, 4, and 6 kJ/m 2) significantly up-regulated the examined genes (e.g. RPA, DNA-PK, Ku70, and Ku80). For the time course study, RPA genes showed immediate up-regulation but returned to basal or lower expression levels compared to the control 3 h after UV-B exposure. The DNA-PK and Ku70/80 genes significantly increased, indicating that they may be involved in repairing processes against a low dose of UV-B exposure (2 kJ/m 2). At the basal level, the hsp90α1 gene showed the highest expression, and followed by hsp10, hsp30, hsp60, and hsc70, and hsp90β in adults (w/o egg). In eggs, the hsp10 gene was expressed the highest, and followed by hsp30, hsp27, hsp90α1, and hsp60 genes. In real-time RT-PCR array on rotifer hsp genes, low doses of UV-B radiation (2 and 4 kJ/m 2) showed up-regulation of several hsp genes but most of the hsp genes showed down-regulation at 8 kJ/m 2 and higher, indicating that significant Hsp-mediated cellular damage already occurred at low doses. For the time course study of four hsp genes ( hsp20, hsp27, hsp70, hsp90α1), they showed a significant correlation for UV-B radiation (2 kJ/m 2). In this paper, we demonstrated that UV-B radiation would affect growth retardation with up- or down-regulation of some important genes in DNA replication, repair process, and chaperoning. This finding provides a better understanding of molecular mechanisms involved in UV-B-mediated cellular damage in the rotifer, Brachionus sp.