KAUST Global Collaborative Research Program, Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong SAR.
J Exp Zool B Mol Dev Evol. 2010 Jul 15;314(5):390-402. doi: 10.1002/jez.b.21344.
Hydroides elegans is a common marine fouling organism in most tropical and subtropical waters. The life cycle of H. elegans includes a planktonic larval stage in which swimming larvae normally take 5 days to attain competency to settle. Larval metamorphosis marks the beginning of its benthic life; however, the endogenous molecular mechanisms that regulate metamorphosis remain largely unknown. In this study, a PCR-based suppressive subtractive hybridization (SSH) library was constructed to screen the genes expressed in competent larvae but not in precompetent larvae. Among the transcripts isolated from the library, 21 significantly matched sequences in the GenBank. Many of these isolated transcripts have putative roles in the reactive oxygen species (ROS) signal transduction pathway or in response to ROS stress. A putative novel p38 mitogen-activated protein kinase (MAPK), which was also isolated with SSH screen, was then cloned and characterized. The MAPK inhibitors assay showed that both p38 MAPK inhibitors SB202190 and SB203580 effectively inhibited the biofilm-induced metamorphosis of H. elegans. A cell stressors assay showed that H(2)O(2) effectively induced larval metamorphosis of H. elegans, but the inductivity of H(2)O(2) was also inhibited by both SB inhibitors. The catalase assay showed that the catalase could effetely inhibit H. elegans larvae from responding to inductive biofilm. These results showed that the p38 MAPK-dependent pathway plays critical role in controlling larval metamorphosis of the marine polychaete H. elegans, and the reactive oxygen radicals produced by biofilm could be the cue inducing larval metamorphosis.
美丽鳞沙蚕是热带和亚热带海域最常见的海洋污损生物之一。美丽鳞沙蚕的生命周期包括浮游幼虫阶段,在此阶段游动的幼虫通常需要 5 天才能达到定居的能力。幼虫变态标志着其底栖生活的开始;然而,调节变态的内源性分子机制在很大程度上仍然未知。在这项研究中,构建了基于 PCR 的抑制性消减杂交(SSH)文库,以筛选在有能力幼虫中表达但不在无能力幼虫中表达的基因。从文库中分离出的转录本中,有 21 个显著匹配的序列在 GenBank 中。这些分离出的转录本中有许多具有参与活性氧(ROS)信号转导途径或对 ROS 应激做出反应的潜在作用。一种与 SSH 筛选分离出的假定新型 p38 丝裂原活化蛋白激酶(MAPK)也被克隆并进行了特征描述。MAPK 抑制剂测定表明,p38 MAPK 抑制剂 SB202190 和 SB203580 均可有效抑制美丽鳞沙蚕生物膜诱导的变态。细胞应激物测定表明,H 2 O 2 可有效诱导美丽鳞沙蚕幼虫变态,但两种 SB 抑制剂也可抑制 H 2 O 2 的诱导性。过氧化氢酶测定表明,过氧化氢酶可有效抑制美丽鳞沙蚕幼虫对诱导性生物膜的反应。这些结果表明,p38 MAPK 依赖性途径在控制海洋多毛类美丽鳞沙蚕幼虫变态中起着关键作用,生物膜产生的活性氧自由基可能是诱导幼虫变态的信号。