title: DNA microarray analysis on gene candidates possibly related to tetrodotoxin accumulation in pufferfish.
abstract: Pufferfish accumulate tetrodotoxin (TTX) at high levels in liver and ovary through the food chain. However, the mechanisms underlying TTX toxification in pufferfish have been poorly understood. In order to search gene candidates involved in TTX accumulation in the torafugu pufferfish Takifugu rubripes, a custom 4x44k oligonucleotide microarray slide was designed by the Agilent eArray program using oligonucleotide probes of 60 bp in length referring to 42,724 predicted transcripts in the publicly available Fugu genome database. DNA microarray analysis was performed with total RNA samples from the livers of two toxic wild specimens in comparison with those from a nontoxic wild specimen and two nontoxic cultured specimens. The mRNA levels of 1108 transcripts were more than 2-fold higher in the toxic specimens than in the nontoxic specimens. The levels of 613 transcripts were remarkably high, and 16 transcripts encoded by 9 genes were up-regulated more than 10-fold. These genes included those encoding forming structural filaments (keratins) and those related to vitamin D metabolism and immunity. It was also noted that the levels of the transcripts encoding serpin peptidase inhibitor clade C member 1, coagulation factor X precursor, complement C2, C3, C5, C8 precursors, and interleukin-6 receptor were high in the toxic liver samples.
authors.affiliation: Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan.
authors.forename: Holger
authors.initials: H
authors.lastname: Feroudj
authors.affiliation: Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Matsumoto
authors.affiliation: Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Kurosu
authors.affiliation: Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan.
authors.forename: Gen
authors.initials: G
authors.lastname: Kaneko
authors.affiliation: Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Ushio
authors.affiliation: Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Minato, Tokyo 108-8477, Japan.
authors.forename: Katsuaki
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Minato, Tokyo 108-8477, Japan.
authors.forename: Hidehiro
authors.initials: H
authors.lastname: Kondo
authors.affiliation: Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Minato, Tokyo 108-8477, Japan.
authors.forename: Ikuo
authors.initials: I
authors.lastname: Hirono
authors.affiliation: Department of Food Science and Technology, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Minato, Tokyo 108-8477, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Nagashima
authors.affiliation: Kanagawa Prefecture Fisheries Research Center, Jogashima, Miura, Kanagawa 238-0237, Japan.
authors.forename: Seiji
authors.initials: S
authors.lastname: Akimoto
authors.affiliation: Kanagawa Prefecture Fisheries Research Center, Jogashima, Miura, Kanagawa 238-0237, Japan.
authors.forename: Kazushige
authors.initials: K
authors.lastname: Usui
authors.affiliation: Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan.
authors.forename: Shigeharu
authors.initials: S
authors.lastname: Kinoshita
authors.affiliation: Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan.
authors.forename: Shuichi
authors.initials: S
authors.lastname: Asakawa
authors.affiliation: Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Kodama
authors.affiliation: Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan; School of Marine Biosciences, Kitasato University, Minami, Sagamihara, Kanagawa 252-0373, Japan. Electronic address: swatabe@kitasato-u.ac.jp.
authors.forename: Shugo
authors.initials: S
authors.lastname: Watabe
mesh_terms: D000818:Animals; D000990:Antithrombin III / Q000378:metabolism; D003165:Complement System Proteins / Q000378:metabolism; D005170:Factor X / Q000378:metabolism; D005400:Fishes, Poisonous / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D007564:Japan; D007633:Keratins / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D015345:Oligonucleotide Probes; D019947:Receptors, Interleukin-6 / Q000378:metabolism; D023721:Tetraodontiformes / Q000235:genetics* / Q000378:metabolism; D013779:Tetrodotoxin / Q000378:metabolism* / Q000493:pharmacokinetics*; D014807:Vitamin D / Q000378:metabolism
chemical_list: D015345:Oligonucleotide Probes; D019947:Receptors, Interleukin-6; D014807:Vitamin D; D013779:Tetrodotoxin; D007633:Keratins; D000990:Antithrombin III; D005170:Factor X; D003165:Complement System Proteins
keywords: APP; DNA microarray; Gene expression; IL-6; Keratin; LC–FLD; Pufferfish; RAP RT–PCR; SSH; TTX; Takifugu rubripes; Tetrodotoxin; acute phase protein; fibrinogen-like protein; flp; interleukin-6; liquid chromatography–fluorescence detection; mRNA arbitrarily primed reverse transcription–polymerase chain reaction; suppression subtractive hybridization; tetrodotoxin