Dynamic Alterations in DNA Methylation Precede Tris(1,3-dichloro-2-propyl)phosphate-Induced Delays in Zebrafish Epiboly.
Dynamic Alterations in DNA Methylation Precede Tris(1,3-dichloro-2-propyl)phosphate-Induced Delays in Zebrafish Epiboly.
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DOI：
10.1021/acs.estlett.7b00332
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发表时间：
2017-09-12
影响因子：
10.9
通讯作者：
Volz DC
中科院分区：
文献类型：
--
作者：
Kupsco A;Dasgupta S;Nguyen C;Volz DC
磷酸三（1，3-二氯-2-丙基）酯（TDCIPP）是一种有机磷酸盐阻燃剂，可影响斑马鱼的表皮-这种效应可能与全基因组低甲基化有关。本研究以斑马鱼为模型，目的是：（1）量化TDCIPP对表皮的浓度依赖性影响;（2）确定与叶酸（FA）（一种甲基供体）共同暴露是否减轻TDCIPP诱导的影响;和（3）使用10个先前鉴定的TDCIPP易感基因座，依赖于亚硫酸氢盐扩增子测序（BSAS）来监测在存在或不存在FA的情况下跨多个TDCIPP浓度的CpG甲基化动态。在存在或不存在ImM FA的情况下，将胚胎从受精后0.75 h（hpf）暴露于TDCIPP至2、4、6或24 hpf。虽然TDCIPP延迟epiboly达3小时6 hpf和诱导畸形24 hpf，FA是无法减轻TDCIPP诱导的影响，在所有阶段的评价。此外，虽然使用5-甲基胞嘧啶（5-mC）DNA ELISA检测到整体甲基化没有差异，但BSAS显示TDCIPP诱导的CpG甲基化效应依赖于浓度和发育阶段，并且尽管持续暴露，但对甲基化的早期效应不会持续存在。我们的研究结果表明，TDCIPP延迟斑马鱼epiboly，一个表型，这是由复杂的，动态的DNA甲基化改变之前。
Tris(1,3-dichloro-2-propyl)phosphate (TDCIPP) is an organophosphate flame retardant that impacts zebrafish epiboly – an effect that may be associated with genome-wide hypomethylation. Using zebrafish as a model, the objectives of this study were to (1) quantify concentration-dependent impacts of TDCIPP on epiboly; (2) determine whether co-exposure with folic acid (FA) – a methyl donor – mitigates TDCIPP-induced impacts; and (3) using ten previously identified TDCIPP-susceptible loci, rely on bisulfite amplicon sequencing (BSAS) to monitor CpG methylation dynamics across multiple TDCIPP concentrations in the presence or absence of FA. Embryos were exposed to TDCIPP from 0.75 h post-fertilization (hpf) to 2, 4, 6, or 24 hpf in the presence or absence of 1 mM FA. Although TDCIPP delayed epiboly up to 3 h by 6 hpf and induced malformations by 24 hpf, FA was unable to mitigate TDCIPP-induced effects at all stages evaluated. Moreover, while no differences in global methylation were detected using a 5-methylcytosine (5-mC) DNA ELISA, BSAS revealed that TDCIPP-induced effects on CpG methylation were dependent on concentration and developmental stage, and that early effects on methylation do not persist despite continuous exposure. Our findings demonstrate that TDCIPP delays zebrafish epiboly, a phenotype that is preceded by complex, dynamic alterations in DNA methylation.
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影响因子：
2.7
作者：
Martin, CC;Laforest, L;Ekker, M
通讯作者：
Ekker, M
影响因子：
2.9
作者：
Jarema KA;Hunter DL;Shaffer RM;Behl M;Padilla S
通讯作者：
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通过下一代测序进行靶向DNA甲基化分析。
DOI：
10.3791/52488
发表时间：
2015-02-24
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
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通讯作者：
Freeman WM
影响因子：
2.5
作者：
KIMMEL, CB;BALLARD, WW;SCHILLING, TF
通讯作者：
SCHILLING, TF
影响因子：
3.8
作者：
Vliet SM;Ho TC;Volz DC
通讯作者：
Volz DC