title: Alteration of development and gene expression induced by in ovo-nanoinjection of 3-hydroxybenzo[c]phenanthrene into Japanese medaka (Oryzias latipes) embryos.
abstract: Benzo[c]phenanthrene (BcP) is a highly toxic polycyclic aromatic hydrocarbon (PAHs) found throughout the environment. In fish, it is metabolized to 3-hydroxybenzo[c]phenanthrene (3-OHBcP). In the present study, we observed the effects of 1nM 3-OHBcP on the development and gene expression of Japanese medaka (Oryzias latipes) embryos. Embryos were nanoinjected with the chemical after fertilization. Survival, developmental stage, and heart rate of the embryos were observed, and gene expression differences were quantified by messenger RNA sequencing (mRNA-Seq). The exposure to 1nM 3-OHBcP accelerated the development of medaka embryos on the 1st, 4th, and 6th days post fertilization (dpf), and increased heart rates significantly on the 5th dpf. Physical development differences of exposed medaka embryos were consistent with the gene expression profiles of the mRNA-Seq results for the 3rd dpf, which show that the expression of 780 genes differed significantly between the solvent control and 1nM 3-OHBcP exposure groups. The obvious expression changes in the exposure group were found for genes involved in organ formation (eye, muscle, heart), energy supply (ATPase and ATP synthase), and stress-response (heat shock protein genes). The acceleration of development and increased heart rate, which were consistent with the changes in mRNA expression, suggested that 3-OHBcP affects the development of medaka embryos. The observation on the developmental stages and heart beat, in ovo-nanoinjection and mRNA-Seq may be efficient tools to evaluate the effects of chemicals on embryos.
authors.affiliation: Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan.
authors.forename: Kun
authors.initials: K
authors.lastname: Chen
authors.affiliation: Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Tsutsumi
authors.affiliation: Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan.
authors.forename: Shuhei
authors.initials: S
authors.lastname: Yoshitake
authors.affiliation: Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan.
authors.forename: Xuchun
authors.initials: X
authors.lastname: Qiu
authors.affiliation: Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan; School of Environment and Safety Engineering, Jiangsu University, Zhenjiang 212013, PR China.
authors.forename: Hai
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Biology, Osaka Medical College, Takatsuki-shi 569-8686, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Hashiguchi
authors.affiliation: Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Honda
authors.affiliation: Laboratory of Molecular Gene Technology, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan.
authors.forename: Kosuke
authors.initials: K
authors.lastname: Tashiro
authors.affiliation: Center for Marine Environmental Studies, Ehime University, 2-5 Bunkyo-cho, Matsuyama 790-8577, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Nakayama
authors.affiliation: National Research Institute of Fisheries and Environment of Inland Sea, Japan Fisheries Research and Education Agency, Maruishi 2-17-5, Hatsukaichi-shi, Hiroshima 739-0452, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Hano
authors.affiliation: Institute of Nature and Environmental Technology, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
authors.forename: Nobuo
authors.initials: N
authors.lastname: Suzuki
authors.affiliation: Institute of Nature and Environmental Technology, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
authors.forename: Kazuichi
authors.initials: K
authors.lastname: Hayakawa
authors.affiliation: Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Shimasaki
authors.affiliation: Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan. Electronic address: yoshima@agr.kyushu-u.ac.jp.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Oshima
mesh_terms: D000818:Animals; D004625:Embryo, Nonmammalian / Q000187:drug effects* / Q000502:physiology; D047108:Embryonic Development / Q000187:drug effects* / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D007267:Injections; D008297:Male; D009990:Oryzias / Q000502:physiology*; D010616:Phenanthrenes / Q000378:metabolism / Q000633:toxicity*; D012333:RNA, Messenger / Q000378:metabolism; D011897:Random Allocation; D018675:Toxicity Tests; D059467:Transcriptome / Q000187:drug effects*; D014874:Water Pollutants, Chemical / Q000378:metabolism / Q000633:toxicity*
chemical_list: D010616:Phenanthrenes; D012333:RNA, Messenger; D014874:Water Pollutants, Chemical; C002615:benzo(c)phenanthrene
keywords: Developmental stages; Heart rate; In ovo-nanoinjection; Medaka embryo; PAHs; mRNA-Seq