Effect of fluorescent whitening agent on the transcription of cell damage‐related genes in zebrafish embryos

Abstract

7-Diethylamino-4-methylcoumarin (DEMC) is a fluorescent whitening agent (FWAs). There have been some studies on DEMC's protective effects against biological activity but there are few papers about the in vivo toxicity of DEMC. In this study, we used wild-type zebrafish embryos 3 days post fertilization (dpf). Test solutions with DEMC concentrations were negative control (without vehicle), 0 (with vehicle, 0.01% v/v ethanol), 0.25, 0.5, 0.75, 1.0, 1.25, 1.5 and 2 ppm. Embryos and larvae were counted for survival rate and hatching rate. Heart rates were also counted at 2.5 and 3.0 dpf. At 3.0 dpf, quantitative RT-PCR was performed with some samples (0, 0.25, 0.75 and 1.25 ppm) to determine the toxic effect to DEMC by detecting the expression levels of toxic-responsive genes. We used 11 genes, which included oxidative stress-related genes [sod(Mn), sod(Cu,Zn) and hsp70], mitochondrial metabolism-related genes (coxI, pyc, cyt and cyclinG1) and apoptosis-related genes (c-jun, bcl2, bax and p53). High-concentration DEMC-treated groups showed significant different survival rate, hatching rate and heart rate compared with low-concentration DEMC-treated groups. The LC50 of this chemical, 0.959 ppm, was calculated. We also confirmed that some genes in the DEMC exposure groups showed significantly up-regulations in expression levels compared with control groups. We concluded that the fluorescence agent, DEMC, has possible developmental toxicities and alteration effect of gene expression, which are related to oxidative stress, mitochondrial metabolism and apoptosis in zebrafish embryos.

Brain Intraventricular Injection of Amyloid-β in Zebrafish Embryo Impairs Cognition and Increases Tau Phosphorylation, Effects Reversed by Lithium

“…The primers sequences were described previously by Tang [30] for constitutive genes: b-actin1 F:5′-CGAGCTGTCTTCCCATCCA-3′, R:5′-TCACC-AACGTZGCTGTCTTTCTG-3′; ef1a F:5′-CTGGAGGCCAGCTCAAACAT–3′, R:5′-ATCAAGAAGAGTAGTACCGCTAGCATTAC-3′; and rpl13a F:5′-TCT-GGAGGACTGTAAGAGGTATGC-3′, R:5′-AGACGCACAATCTTGAGAGCAG-3′. The target genes were previously described by Jung et al [31]: p53 F:5′-CTATAAGAAGTCCGAGCATGTGG-3′, R:5′-GGTTTTGGTCTCTTGGTCTTCT-3′; bax-a F:5′-GAGCTGCACTTCTCAACAACTTT-3′, R:5′-CTGGTTGAAATAG-CCTTGATGAC-3′ and bcl-2 F:5′-TTGTGGAGAAATACCTCAAGCAT-3′, R:5′-GAGTCTCTCTGCTGACCGTACAT-3′. Amplification and dissociation curves generated by the software were used for gene expression analysis.…”

Exposure time to caffeine affects heartbeat and cell damage‐related gene expression of zebrafish Danio rerio embryos at early developmental stages

“…Our results showed significant changes in all tested genes. Several studies reported an induced expression of these genes after exposure to different genotoxic compounds (Gonzalez et al, 2006;Jung et al, 2011). HSP70 is a molecular biomarker of cellular stress that induces denaturation of other proteins (Sanders, 1990;Lewis et al, 1999) because it is involved in protecting and defending cells from environmental insults (Sanders, 1990); its induction is much more responsive than traditional indices of contaminant effects (Feder and Hofmann, 1999).…”

“…The up-regulation expression of Bcl2 shows the activation of apoptotic cell death. However, many researchers have presented not expression data for both Bax and Bcl2 genes but the ratio of Bax/Bcl2 as an indicator for cell apoptosis (Jung et al, 2011;Basu and Haldar, 1998). The up-regulation of Bax/ Bcl2 ratio shows cell apoptosis (Pepper et al, 1997;Pavlovic et al, 2006;Salakou et al, 2007;Jin et al, 2011).…”

Functional expressions of adenosine triphosphate‐binding cassette transporters during the development of zebrafish embryos and their effects on the detoxification of cadmium chloride and β‐naphthoflavone

“…Fertilized eggs were obtained from matured healthy male and female zebrafish according to Jung et al . () with slight modification. Two male zebrafish and one female zebrafish were placed separately with a barrier in a spawning box, and a mesh bottom was used to prevent the eggs from being cannibalized.…”