Effects of β-diketone antibiotics on F1-zebrafish (Danio rerio) based on high throughput miRNA sequencing under exposure to parents

Abstract

The toxicity of β-diketone antibiotics (DKAs), a class of ''pseudo-persistent'' environmental pollutants, to F0-zebrafish (Danio rerio) was investigated using 7-dpf F1-zebrafish miRNA sequencing and bioinformatics analyses. Based on relative expression, 47, 134 and 118 of 193 mature miRNAs were differentially expressed between control vs 6.25 mg/L, control vs 12.5 mg/L and 6.25 vs 12.5 mg/L treatments, respectively. Utilizing three databases, 2523 potential target genes were predicted, and they were assigned to 19 high-abundance KEGG pathways and 20 functional categories by COG analysis. Among 11 significantly differential expression and high-abundance miRNAs, the expression levels for 7 miRNAs (miR-144, -124, -499, -125b, -430b, -430c and -152) assessed by qRT-PCR were consistent with those determined by sRNA-seq. A potential network was plotted between 11 miRNAs and their target genes based on differential expression and binding effectiveness. The high degree of connectivity between miRNA-gene pairs suggests that these miRNAs play critical roles in zebrafish development. The expression of miR-124 and miR-499 in whole-mount in situ hybridization was in general agreement with those from qRT-PCR and miRNA-seq and were DKA concentration-dependent. DKA exposure induced severe histopathological changes and damage in F0-zebrafish ovary tissue, as reflected by an increased number of early developmental oocytes, irregular cell distribution, decreased yolk granules, cytoplasmic shrinkage, cell lysis in mature oocytes, and dissolution of internal corona radiata. Chronic DKA exposure affected reproduction of F0-zebrafish and development of F1-zebrafish. These observations demonstrate the toxic effect transfer relation across parent and their offspring, and enhance our understanding of drug-induced diseases.

Lipid metabolism disorder induced by up-regulation of miR-125b and miR-144 following β-diketone antibiotic exposure to F0-zebrafish (Danio rerio)

“…Sequencing data showed that target genes regulated by high-abundance and positive differentially expressed miRNAs were involved in cancer, hepatitis B, cell cycle, Salmonella infection, metabolism of fatty acid synthesis, etc. (Zheng et al, 2016). However, these results were based on bioinformatics prediction from sequencing data and required further verification.…”

“…The miRNA sequencing and bioinformatics analyses for 5-dpf F1zebrafish previously identified a high abundance of positive differentially expressed miRNA-144 and miRNA-125b for each DKA treatment based on the criteria of Log 2 (|fold-change|)≥ 1.0, per million fragments mapped (FPKM)≥ 100 and p < 0.05 (Zheng et al, 2016). To better understand the biological functions of miRNA-144 and miRNA-125b, we predicted their potential target genes using TargetScan software.…”

Zebrafish as a Screening Model to Study the Single and Joint Effects of Antibiotics

“…Based on high-throughput screening and histopathological observations, joint DKA exposure on larval and adult zebrafish resulted in abnormal expression of a large number of micro-RNAs; some long non-coding RNAs and their regulating target genes; and a series of physiological changes in tissues and organs, especially those related to the nervous, immune and reproductive systems [ 32 , 112 , 113 , 114 , 115 , 116 ]. For example, the joint FQ and TC exposure induced severe histopathological changes and damage in the zebrafish eye, brain, hepatic and spleen tissues ( Figure 2 ), including vacuolation of interstitial cells, photoreceptor cell cysts, reduced number of neurons, glial cell proliferation, formation of glial scars, hepatic parenchyma vacuolar degeneration and brown metachromatic granules.…”

“…For example, the joint FQ and TC exposure induced severe histopathological changes and damage in the zebrafish eye, brain, hepatic and spleen tissues ( Figure 2 ), including vacuolation of interstitial cells, photoreceptor cell cysts, reduced number of neurons, glial cell proliferation, formation of glial scars, hepatic parenchyma vacuolar degeneration and brown metachromatic granules. On the other hand, the female reproductive system suffered serious damage after chronic exposure to DKAs, as reflected by the increased number of early developmental oocytes, irregular cell distribution, different cellular morphology, decreased yolk granules, cytoplasmic shrinkage and cell lysis in mature oocytes [ 32 ]. Moreover, Wang et al [ 114 ] showed the adverse reproductive effects of β-diketone antibiotics in female zebrafish, along with an effect transfer relation across parents and their offspring.…”

“…Most of the research papers on this topic have reported that the zebrafish health or/and its capacity to respond to environmental changes are affected following both acute and chronic exposure to antibiotics. The toxicological effects of single and mixed antibiotics in zebrafish were studied by proteomic [ 30 ] and transcriptome sequencing [ 31 , 32 ], examination of behavioral activities [ 28 , 33 , 34 ] and biomarkers [ 35 , 36 ], histopathological analysis of various tissues [ 30 , 37 ], fish lethality assays (LC 50 ) [ 22 , 38 , 39 ] and following the fish morphological changes [ 40 ]. Antibiotics were found to interfere with the bacterial communities of zebrafish gut and with fish development, causing malformations, such as hatching delay, shorter body length, yolk sac edema, curved body axis and pericardial edema [ 23 , 25 , 41 , 42 , 43 , 44 ].…”