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河豚肌肉注射毒素后肝脏差异表达基因谱

Differential gene expression profile in the liver of the marine puffer fish Takifugu rubripes induced by intramuscular administration of tetrodotoxin.

机构信息

Department of Food Science and Technology, Tokyo University of Marine Science and Technology, 4-5-7 Konan, Minato, Tokyo 108-8477, Japan.

出版信息

Toxicon. 2011 Feb;57(2):304-10. doi: 10.1016/j.toxicon.2010.12.007. Epub 2010 Dec 21.

DOI:10.1016/j.toxicon.2010.12.007
PMID:21172371
Abstract

Marine puffer fish accumulate a high level of tetrodotoxin (TTX) in the liver and ovary, but the underlying mechanism of this toxification is unclear. To elucidate the genes related to toxification of the marine puffer fish, we examined the hepatic gene expression profile of the marine puffer fish Takifugu rubripes by suppression subtractive hybridization in response to the intramuscular administration of 0.50 mg TTX/kg body weight into the caudal muscle. The accumulation of TTX in the liver reached 68 ± 4% that of the administered dose within 12 h of administration. A total of 1048 clones from the subtracted cDNA libraries were successfully sequenced. The nucleotide sequence of 92 of the 1048 clones was identified as a hepcidin precursor. Reverse transcription-polymerase chain reaction experiments revealed that hepcidin precursors were highly expressed in the TTX-administered group. In addition, complement C3 (31 clones), serotransferrin (30 clones), apolipoprotein A-1 (14 clones), high temperature adaptation protein Wap65-2 (14 clones), complement C7 (12 clones), fibrinogen beta chain (12 clones), and 70 kDa heat-shock protein 4 (11 clones) were obtained. This study confirmed that the intramuscular administration of TTX increases the gene expression of the acute-phase response proteins in the liver of puffer fish T. rubripes.

摘要

河豚鱼肝脏和卵巢中积累了高水平的河豚毒素(TTX),但其毒化的潜在机制尚不清楚。为了阐明河豚鱼毒化相关的基因,我们通过抑制性消减杂交技术,检测了河豚鱼肝脏基因表达谱,河豚鱼在肌肉内注射 0.50mg TTX/kg 体重后,TTX 在 12 小时内在肝脏中的积累达到了给药剂量的 68±4%。从消减 cDNA 文库中成功测序了 1048 个克隆。1048 个克隆中的 92 个核苷酸序列被鉴定为铁调素前体。逆转录聚合酶链反应实验表明,铁调素前体在 TTX 给药组中高度表达。此外,还获得了 31 个补体 C3 克隆、30 个血清转铁蛋白克隆、14 个载脂蛋白 A-1 克隆、14 个高温适应蛋白 Wap65-2 克隆、12 个补体 C7 克隆、12 个纤维蛋白原β链克隆和 11 个 70kDa 热休克蛋白 4 克隆。本研究证实,河豚鱼肌肉内注射 TTX 可增加河豚鱼肝脏中急性期反应蛋白的基因表达。

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