暗纹东方鲀组织内TTX含量与钠离子通道基因表达特征分析
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国家自然科学基金面上项目(31872546,32170514)


Analysis of tetrodotoxin content and voltage-gated sodium channel genes characteristics in the tissues of Takifugu obscurus
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    摘要:

    为了探究暗纹东方鲀(Takifugu obscurus)组织内河鲀毒素(Tetrodotoxin,TTX)含量与含毒组织电压门控钠离子通道基因表达特征。分别采用酶联免疫法(ELISA)测定暗纹东方鲀肝脏、心脏等8个组织TTX含量,实时荧光定量(RT-qPCR)方法检测电压门控钠离子通道α亚基基因家族成员scn1aa,scn1ab,scn4aa,scn4ab,scn5aa,scn5ab,scn8aa,scn8ab(以下简称“电压门控钠离子通道基因”)在各组织中的相对表达特征。结果显示:各组织中TTX含量由高到低为脑(9.521±2.816) μg/g、心脏(4.271±1.129) μg/g、鳃(1.586±0.527) μg/g、肌肉(1.494±0.938) μg/g、肝脏(0.913±0.206) μg/g、皮肤(0.902±0.235) μg/g、肠道(0.894±0.215) μg/g和眼(0.864±0.287) μg/g,脑、心脏组织为弱毒性,肝脏、皮肤、肌肉、眼、肠道、鳃组织为微毒性。RT-qPCR结果显示scn1aa、scn1ab、scn4ab、scn5aa、scn5ab、scn8aa、scn8ab基因均在肝脏组织的相对表达量最高,scn4aa基因仅在肌肉组织表达,皮肤组织仅有scn4ab基因表达,且scn4ab基因在所有检测组织中均有表达,表达具有广泛性。钠离子通道基因在斑马鱼和暗纹东方鲀组织表达特征存在差异。

    Abstract:

    In order to investigate the tetrodotoxin(TTX) content in the tissues of Takifugu obscurus and the expression characteristics of voltage gated sodium channel genes in toxic tissues, this study used enzyme-linked immunosorbent assay (ELISA) to determine the TTX content in 8 tissues, including the liver and heart of Takifugu obscurus. Real time fluorescence quantification (RT-qPCR) was used to detect the relative expression of voltage gated sodium channels α subunit gene family, including scn1aa, scn1ab, scn4aa, scn4ab, scn5aa, scn5ab, scn8aa, and scn8ab in various tissues. The results showed that the TTX content in various tissues ranged from high to low, including brain (9.521 ±2.816) μg/g, heart (4.271 ±1.129) μg/g, gills (1.586 ±0.527) μg/g, muscle (1.494 ±0.938) μg/g, liver (0.913 ±0.206) μg/g, skin (0.902 ±0.235) μg/g, intestine (0.894 ±0.215) μg/g, and eye (0.864 ±0.287) μg/g. Brain and heart tissues were weakly toxic, while liver, skin, muscle, eye, intestine, and gill tissues were slightly toxic. The RT-qPCR results showed that the relative expression levels of scn1aa, scn1ab, scn4ab, scn5aa, scn5ab, scn8aa, and scn8ab genes were the highest in liver tissue. The scn4aa gene was only expressed in muscle tissue, and only the scn4ab gene was expressed in skin tissue. Moreover, the scn4ab gene was expressed in all detected tissue types, indicating widespread expression. It is discovered that there are differences in the expression characteristics of sodium channel genes between zebrafish and Takifugu obscurus tissues.

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王婧,王润,林梦娇,王亚宁,鲍宝龙,龚小玲.暗纹东方鲀组织内TTX含量与钠离子通道基因表达特征分析[J].上海海洋大学学报,2024,33(2):308-316.
WANG Jing, WANG Run, LIN Mengjiao, WANG Yaning, BAO Baolong, GONG Xiaoling. Analysis of tetrodotoxin content and voltage-gated sodium channel genes characteristics in the tissues of Takifugu obscurus[J]. Journal of Shanghai Ocean University,2024,33(2):308-316.

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  • 收稿日期:2023-05-20
  • 最后修改日期:2023-05-24
  • 录用日期:2023-05-24
  • 在线发布日期: 2024-03-19
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