Received: 5 December 2017 Revised: 22 December 2017 Accepted: 27 December 2017
Methylparaben induces malformations and alterations
on apoptosis, oxidant–antioxidant status, ccnd1 and myca
expressions in zebrafish embryos
Perihan Seda Ate¸s
Ünsal Veli Üstünda
Ahmet Ata Alturfan
Department of Biochemistry, Faculty of Den-
tistry, Marmara University, Maltepe, Istanbul,
Department of Biochemistry, Faculty of
Medicine, Medipol University, Kavacık, Istan-
Department of Biochemistry, Cerrahpasa
Medical Faculty, Istanbul University, Istanbul,
Methylparabens (MP) are widely used as preservatives in cosmetics, pharmacy, and food industry.
Although acute toxicity studies in animals indicated that parabens are not significantly toxic, the
effects of chronic exposure under sublethal doses are still unknown and the number of related
studies is limited. Our aim was to evaluate the effects of MP on the development of zebrafish
embryos focusing on development, locomotor activity, oxidant–antioxidant status, apoptosis, and
ccnd1 and myca expressions.The expressions of ccnd1 and myca were determined by RT-PCR.Lipid
peroxidation (LPO), nitric oxide (NO), and glutathione-S-transferase (GST) activities were deter-
mined spectrophotometrically. Apoptosis was determined using acridine orange staining. Loco-
motor activity was measured using touch-evoked movement test. MP exposure increased malfor-
mations, LPO, apoptosis, ccnd1 and myca expressions, and decreased GST activities and NO levels
compared with the control group. Our findings will lead to further understanding of the mecha-
nism of MP toxicity, and merit further research.
apoptosis, ccnd1, methylparaben, myca, oxidant–antioxidant status
Endocrine-disrupting compounds (EDC) have gained popularity as a
popular subject in human health. EDCs are widespread in the aquatic
environment and can cause alterations in development, physiological
homeostasis, and health of vertebrates by acting in estrogenic, antie-
strogenic, androgenic, or antiandrogenic ways.
Zebrafish, Danio rerio,
has been suggested as a model to identify targets as well as modes of
Methyl ester of p-hydroxybenzoic acid (parabens) are widely used
as preservatives in cosmetics, pharmacy, and food. Acute toxicity
studies in animals indicated that parabens are not significantly toxic
through various routes of administration and subchronic and chronic
oral studies indicated that parabens are practically nontoxic.
all these, the effects of chronic exposure under sublethal doses are still
unknown and the number of studies on the effects of methylparaben
(MP) in literature is limited.
Several alkyl parabens have been shown to exert binding affini-
ties to estrogen receptors and MP has been shown to affect
FSH levels, folliculogenesis, and cause histopathological effects.
On the other hand, exposure to MP from early embryonic stage
is a major concern and it has been shown that parabens exist
as high concentration in maternal blood, plasma, and amniotic
Myc, a transcription factor that regulates the expression of cellu-
lar genes, is well known for its participation in many malignant conver-
sions. Cyclin D1 (Ccnd1) activates cyclin dependent kinases CDK4 and
CDK6 and drives cell proliferation.
Analyzing apoptosis and oxida-
tive stress markers is also important to evaluate the relative toxic-
ity of EDCs. Because of its small size and ease of reproduction and
breeding, zebrafish is one of the most commonly used fish species in
xenoestrogenic potential testing in embryonic stage.
The aim of this
study was to evaluate the effects of MP exposure on the development
of zebrafish embryo focusing on the apoptosis, oxidant–antioxidant
parameters, locomotor activity, and myca and ccnd1 expressions.
2 MATERIAL AND METHODS
MP (CAS no: 99-76-3) was purchased from SUPELCO (Bellefonte, PA)
and acetone (67-64-1) was purchased from Merck (Darmstadt, Ger-
many). They were all analytical grade with the highest purity available.
J Biochem Mol Toxicol. 2018;32:e22036.
2018 Wiley Periodicals, Inc. 1of6wileyonlinelibrary.com/journal/jbt