Gene expression profiling in respond to TBT exposure in small abalone Haliotis diversicolor

doi:10.1016/j.fsi.2011.07.004

Fish & Shellfish Immunology

Volume 31, Issue 4, October 2011, Pages 557-563

https://doi.org/10.1016/j.fsi.2011.07.004 Get rights and content

Abstract

In this study, we investigated the gene expression profiling of small abalone, Haliotis diversicolor by tributyltin (TBT) exposure using a cDNA microarray containing 2473 unique transcripts. Totally, 107 up-regulated genes and 41 down-regulated genes were found. For further investigation of candidate genes from microarray data and EST analysis, quantitative real-time PCR was performed at 6 h, 24 h, 48 h, 96 h and 192 h TBT exposure. 26 genes were found to be significantly differentially expressed in different time course, 3 of them were unknown. Some gene homologues like cellulose, endo-beta-1,4-glucanase, ferritin subunit 1 and thiolester containing protein II CG7052-PB might be the good biomarker candidate for TBT monitor. The identification of stress response genes and their expression profiles will permit detailed investigation of the defense responses of small abalone genes.

Highlights

► We investigated the gene expression profiling of small abalone by TBT exposure. ► 107 up-regulated genes and 41 down-regulated genes were found by microarray. ► 26 significantly differentially expressed genes were found by real-time PCR. ► Some gene homologues might be the good biomarker candidate for TBT monitor.

Section snippets

Introductions

Tributyltin (TBT) is a common contaminant of marine and freshwater ecosystems due to its widespread use as an antifouling agent in boat paints. Although a worldwide ban on TBT was imposed in 2003 [1], high concentrations of TBT can still be found in seawater and sediment [2], [3], [4]. Recent studies have revealed extremely high concentrations of organotins in some pristine natural habitats such as the Great Barrier Reef (GBR) and the Antarctic associated with shipping activities. Sediments at

Animals

Small abalones whose shell width was 4.65 ± 0.27 cm and whose weight was 10.07 ± 2.03 g were collected from a commercial farm (Futian, Dongshan, Fujian Province) and maintained in polyethylene tanks for up to 2 weeks until they acclimatized themselves to the new environment. Fresh seawater was changed daily with 100%. Animals were fed with fresh kelp (Laminaria japonica Aresch) while water was being changed. Each tank contained 60 animals in 180 L of aerated and sand filtered seawater at

Microarray data analysis

Based on the criteria selected for microarray analysis (see above), significant differences in gene expression were observed at 96 h between the TBT exposure and control small abalones. At 96 h, 107 up-regulated and 41 down-regulated genes were found to be differentially expressed; among them 62 (42%) genes were unknown genes (Table 1 and Table 2).

Confirmation of microarray by rqRT-PCR

In order to confirm the microarray results, 14 candidate genes from up-regulated genes and 12 candidate genes from down-regulated genes were further

Discussion

It is known that TBT has adverse effects on cellular immune functions like mobility, phagocytosis and lysosomal enzyme activity in invertebrates [33]. Exposure of the oyster to TBT could also enhance the progression of infection by the pathogenic protozoan [47]. In this work, we found that some genes related to immune system such as acid and alkaline phosphatase (ACP and AKP), cathepsins, peptidoglycan recognition protein (PGRP), Alpha-2-macroglobulin, C-type lectin, a disintegrin,

Acknowledgements

The project was support by International Science Foundation (No. A/4727-1), Natural Science Foundation of China (No. 20877034), Innovation Team Foundation of Jimei University (No. 2010A001) and the Fund of Key Lab of Marine Biogenetic Resources (SOA) (No. HY201003). We thank Mr. Ion Beldorth, Department of Chemistry and Biochemistry, Texas State University for his proof reading of the manuscript.