High-throughput RNA sequencing reveals the effects of 2,2′,4,4′ -tetrabromodiphenyl ether on retina and bone development of zebrafish larvae

BMC Genomics

Bingzhi Dong²  Qingshun Zhao¹  Daqiang Yin³  Jing Zhao³  Ting Xu³ 

[1] Model Animal Research Center, MOE Key Laboratory of Model Animal for Disease Study, Nanjing University, Nanjing 210061, China;Post-doctoral Research Station of Civil Engineering, Tongji University, Shanghai 200092, China;Key Laboratory of Yangtze River Water Environment, Ministry of Education, College of Environmental Science and Technology, Tongji University, Shanghai 200092, China

关键词: Visual perception;    Functional enrichment analysis;    Zebrafish larvae;    RNA sequencing;    Polybrominated diphenyl ethers;   

Others  :  1109715
DOI  :  10.1186/s12864-014-1194-5

 received in 2014-08-05, accepted in 2014-12-22,  发布年份 2015

【 摘 要 】

Background

2,2′,4,4′-Tetrabromodiphenyl ether (BDE47) is a prevalent environmental pollutant and has been demonstrated to be a serious toxicant in both humans and animals, but little is known about the molecular mechanism underlying its toxic effect on the early development of vertebrates. BDE47-treated zebrafish larvae were found to present the light-related locomotion reduction in our previous study, therefore, we aimed to use high throughput sequencing to investigate the possible reasons from a transcriptomic perspective.

Results

By exposing zebrafish embryos/larvae to 5 μg/l and 500 μg/l BDE47, we measured the influence of BDE47 on the mRNA expression profiles of zebrafish larvae until 6 days post-fertilization, using Illumina HiSeq 2000 sequencing. Differential expression analysis and gene enrichment analysis respectively revealed that a great number of genes, and gene sets based on two popular terminologies, were affected by the treatment of 500 μg/l BDE47. Among them, BDE47 caused changes in the retinal metabolism and related biological processes involving eye morphogenesis and visual perception, as confirmed by disordered photoreceptor arrangement and thickened bipolar cell layer of larval retina from histological observations. Other altered genes such as pth1a and collaborative cathepsin family exhibited disrupted bone development, which was consistent with the body curvature phenotype. The transcriptome of larvae was not significantly affected by the treatment of 5 μg/l BDE47, as well as the treatment of DMSO vehicle.

Conclusions

Our results suggest that high BDE47 concentrations disrupt the eye and bone development of zebrafish larvae based on both transcriptomic and morphological evidences. The abnormal visual perception may result in the alteration of dark adaption, which was probably responsible for the abnormal larval locomotion. Body curvature arose from enhanced bone resorption because of the intensive up-regulation of related genes. We also proposed the larval retina as a novel potential target tissue for BDE47 exposure.

【 授权许可】

   
2015 Xu et al.; licensee Biomed Central.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Chernyak SM, Rice CP, Quintal RT, Begnoche LJ, Hickey JP, Vinyard BT: Time trends (1983–1999) for organochlorines and polybrominated diphenyl ethers in rainbow smelt (Osmerus mordax) from Lakes Michigan, Huron, and Superior, USA. Environ Toxicol Chem 2005, 24:1632-1641.
• [2]Carlsson P, Herzke D, Wedborg M, Gabrielsen GW: Environmental pollutants in the Swedish marine ecosystem, with special emphasis on polybrominated diphenyl ethers (PBDE). Chemosphere 2011, 82:1286-1292.
• [3]Birnbaum LS, Staskal DF: Brominated flame retardants: cause for concern? Environ Health Persp 2004, 112:9-17.
• [4]Staskal DF, Diliberto JJ, DeVito MJ, Birnbaum LS: Toxicokinetics of BDE 47 in female mice: Effect of dose, route of exposure, and time. Toxicol Sci 2005, 83:215-223.
• [5]Darnerud PO: Toxic effects of brominated flame retardants in man and in wildlife. Environ Int 2003, 29:841-853.
• [6]Richardson VM, Staskal DF, Ross DG, Diliberto JJ, DeVito MJ, Birnbaum LS: Possible mechanisms of thyroid hormone disruption in mice by BDE 47, a major polybrominated diphenyl ether congener. Toxicol Appl Pharmacol 2008, 226:244-250.
• [7]Costa LG, Giordano G: Developmental neurotoxicity of polybrominated diphenyl ether (PBDE) flame retardants. Neurotoxicology 2007, 28:1047-1067.
• [8]Scholz S: Zebrafish embryos as an alternative model for screening of drug-induced organ toxicity. Arch Toxicol 2013, 87:767-769.
• [9]Huang QS, Dong SJ, Fang C, Wu XL, Ye T, Lin Y: Deep sequencing-based transcriptome profiling analysis of Oryzias melastigma exposed to PFOS. Aquat Toxicol 2012, 120:54-58.
• [10]Olsvik PA, Berg V, Lyche JL: Transcriptional profiling in burbot (Lota lota) from Lake Mjosa-A Norwegian Lake contaminated by several organic pollutants. Ecotox Environ Safe 2013, 92:94-103.
• [11]Zhao J, Xu T, Yin DQ: Locomotor activity changes on zebrafish larvae with different 2,2′,4,4′-tetrabromodiphenyl ether (PBDE-47) embryonic exposure modes. Chemosphere 2014, 94:53-61.
• [12]Chen LG, Huang YB, Huang CJ, Hu B, Hu CY, Zhou BS: Acute exposure to DE-71 causes alterations in visual behavior in zebrafish larvae. Environ Toxicol Chem 2013, 32:1370-1375.
• [13]Vesterlund L, Jiao H, Unneberg P, Hovatta O, Kere J. The zebrafish transcriptome during early development. BMC Dev Biol. 2011;11: 10.1186/1471-213X-11-30
• [14]Dillies MA, Rau A, Aubert J, Hennequet-Antier C, Jeanmougin M, Servant N, Keime C, Marot G, Castel D, Estelle J, Guernec G, Jagla B, Jouneau L, Laloe D, Le Gall C, Schaeffer B, Le Crom S, Guedj M, Jaffrezic F: A comprehensive evaluation of normalization methods for Illumina high-throughput RNA sequencing data analysis. Brief Bioinform 2013, 14:671-683.
• [15]Stricher F, Macri C, Ruff M, Muller S: HSPA8/HSC70 chaperone protein Structure, function, and chemical targeting. Autophagy 2013, 9:1937-1954.
• [16]Bromme D, Li ZQ, Barnes M, Mehler E: Human cathepsin V functional expression, tissue distribution, electrostatic surface potential, enzymatic characterization, and chromosomal localization. Biochemistry 1999, 38:2377-2385.
• [17]Yin J, Shine L, Raycroft F, Deeti S, Reynolds A, Ackerman KM, et al. Inhibition of the Pim1 Oncogene Results in Diminished Visual Function. PLoS One 2013;7. 10.1371/journal.pone.0052177
• [18]Thomas R, McHale CM, Lan Q, Hubbard AE, Zhang LP, Vermeulen R, Li GL, Rappaport SM, Yin SN, Rothman N, Smith MT: Global gene expression response of a population exposed to benzene: A pilot study exploring the use of RNA-sequencing technology. Environ Mol Mutagen 2013, 54:566-573.
• [19]Turner C, Sawle A, Fenske M, Cossinsy A: Implications of the solvent vehicles dimethylformamide and dimethylsulfoxide for establishing transcriptomic endpoints in the zebrafish embryo toxicity test. Environ Toxicol Chem 2012, 31:593-604.
• [20]Wang YP, Hong Q, Qin DN, Kou CZ, Zhang CM, Guo M, Guo XR, Chi X, Tong ML: Effects of embryonic exposure to polychlorinated biphenyls on zebrafish (Danio rerio) retinal development. J Appl Toxicol 2012, 32:186-193.
• [21]Chen LG, Huang CJ, Hu CY, Yu K, Yang LH, Zhou BS: Acute exposure to DE-71: Effects on locomotor behavior and developmental neurotoxicity in zebrafish larvae. Environ Toxicol Chem 2012, 31:2338-2344.
• [22]Chen XJ, Huang CJ, Wang XC, Chen JF, Bai CL, Chen YH, Chen XP, Dong QX, Yang DR: BDE-47 disrupts axonal growth and motor behavior in developing zebrafish. Aquat Toxicol 2012, 120–121:35-44.
• [23]Padilla S, Hunter DL, Padnos B, Frady S, MacPhail RC: Assessing locomotor activity in larval zebrafish: Influence of extrinsic and intrinsic variables. Neurotoxicol Teratol 2011, 33:624-630.
• [24]Lema SC, Schultz IR, Scholz NL, Incardona JP, Swanson P: Neural defects and cardiac arrhythmia in fish larvae following embryonic exposure to 2,2′,4,4′-tetrabromodiphenyl ether (PBDE 47). Aquat Toxicol 2007, 82:296-307.
• [25]Ala-Laurila P, Cornwall MC, Crouch RK, Kono M: The action of 11-cis-retinol on cone opsins and intact cone photoreceptors. J Biol Chem 2009, 284:10422-10432.
• [26]Saari JC: Vitamin A Metabolism in Rod and Cone Visual Cycles. Annu Rev Nutr 2012, 32:125-146.
• [27]Furukawa T, Morrow EM, Cepko CL: Crx, a novel otx-like homeobox gene, shows photoreceptor-specific expression and regulates photoreceptor differentiation. Cell 1997, 91:531-541.
• [28]Babb SG, Kotradi SM, Shah B, Chiappini-Williamson C, Bell LN, Schmeiser G, Chen E, Liu Q, Marrs JA: Zebrafish R-cadherin (Cdh4) controls visual system development and differentiation. Dev Dynam 2005, 233:930-945.
• [29]Murray TM, Rao LG, Divieti P, Bringhurst FR: Parathyroid hormone secretion and action: Evidence for discrete receptors for the carboxyl-terminal region and related biological actions of carboxyl-terminal ligands. Endocr Rev 2005, 26:78-113.
• [30]Delaisse JM, Eeckhout Y, Vaes G: In vivo and in vitro evidence for the involvement of cysteine proteinases in bone resorption. Biochem Bioph Res Co 1984, 125:441-447.
• [31]Alt B, Reibe S, Feitosa NM, Elsalini OA, Wendl T, Rohr KB: Analysis of origin and growth of the thyroid gland in zebrafish. Dev Dynam 2006, 235:1872-1883.
• [32]Holtrop ME, Raisz LG, Simmons HA: The effects of parathyroid hormone, colchicine, and calcitonin on the ultrastructure and the activity of osteoclasts in organ culture. J Cell Biol 1974, 60:346-355.
• [33]Wang LK, Feng ZX, Wang X, Wang XW, Zhang XG: DEGseq: an R package for identifying differentially expressed genes from RNA-seq data. Bioformatics 2010, 26:136-138.
• [34]Zhou XX, Wu WW, Li H, Cheng YM, Wei N, Zong J, Feng XY, Xie ZQ, Chen D, Manley JL, Wang H, Feng Y: Transcriptome analysis of alternative splicing events regulated by SRSF10 reveals position-dependent splicing modulation. Nucleic Acids Res 2014, 42:4019-4030.