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甲壳质酶基因家族在克氏原螯虾抵抗重金属胁迫和体色形成中的表达与功能分析。

Expression and function analysis of crustacyanin gene family involved in resistance to heavy metal stress and body color formation in Exopalaemon carinicauda.

机构信息

Jiangsu Key Laboratory of Marine Bioresources and Environment, Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang, China.

Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Ocean University, Lianyungang, China.

出版信息

J Exp Zool B Mol Dev Evol. 2021 Jun;336(4):352-363. doi: 10.1002/jez.b.23025. Epub 2021 Jan 19.

Abstract

Crustacyanin has the function of binding astaxanthin which is the best antioxidant, and plays an important role in the body color variation of crustaceans. To investigate the causes of body color variation of the ridgetail white prawn, Exopalaemon carinicauda, the present study obtained four subtypes of crustacyanin gene: C1, C2, A1, and A2. Based on fluorescence quantitative polymerase chain reaction, lipocalin-C1 is mainly expressed in the eyestalk, lipocalin-C2 is in the ventral nerve cord, and lipocalin-A1 and lipocalin-A2 are in subcutaneous adipose tissues. Under the inhibiting effect of Cd stress, the expression of four subtypes first increases and then decreases within 24 h, and reaches the maximum at 6 or 12 h. RNA interference experiments showed a decrease in the expression of lipocalin genes in subcutaneous adipose tissue for each subtype, with the body color changing from transparent to red, and the dark red spots on the epidermis changing to bright red. Moreover, the blue protein in the subcutaneous adipose tissue largely disappeared, based on the light micrographs. In view of these findings, the crustacyanin gene appears to fulfill some function in the resistance to heavy metal stress and body color formation of E. carinicauda.

摘要

甲壳质蓝蛋白具有结合虾青素(最好的抗氧化剂)的功能,在甲壳类动物体色变化中起着重要作用。为了研究脊尾白虾体色变化的原因,本研究获得了四种甲壳质蓝蛋白基因亚型:C1、C2、A1 和 A2。基于荧光定量聚合酶链反应,脂钙蛋白-C1 主要在眼柄中表达,脂钙蛋白-C2 在腹神经索中表达,脂钙蛋白-A1 和脂钙蛋白-A2 在皮下脂肪组织中表达。在 Cd 胁迫的抑制作用下,四种亚型的表达在 24 小时内先增加后减少,在 6 或 12 小时达到最大值。RNA 干扰实验表明,每个亚型的皮下脂肪组织中脂钙蛋白基因的表达减少,使体色从透明变为红色,表皮上的暗红色斑点变为鲜红色。此外,基于光镜照片,皮下脂肪组织中的蓝色蛋白大量消失。鉴于这些发现,甲壳质蓝蛋白基因似乎在脊尾白虾抵抗重金属胁迫和体色形成中发挥了一定的作用。

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