CgLOX4基因在长牡蛎幼虫发育中的表达特征

胡悦, 冷金源, 孙洁洁, 王玲玲, 宋林生

大连海洋大学学报 ›› 2024, Vol. 39 ›› Issue (1) : 38-47.

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大连海洋大学学报 ›› 2024, Vol. 39 ›› Issue (1) : 38-47. DOI: 10.16535/j.cnki.dlhyxb.2023-120

CgLOX4基因在长牡蛎幼虫发育中的表达特征

  • 胡悦,冷金源,孙洁洁*,王玲玲*,宋林生
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Expression characteristics of CgLOX4 gene in larval development of Pacific oyster (Crassostrea gigas)

  • HU Yue,LENG Jinyuan,SUN Jiejie*,WANG Lingling*,SONG Linsheng
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摘要

为研究长牡蛎(Crassostrea gigasLOX4基因(CgLOX4)在幼虫发育中的表达特征,采用PCR技术扩增CgLOX4的全长cDNA序列,利用荧光定量PCR和整体免疫荧光技术检测CgLOX4在幼虫不同发育时期的表达特征。结果表明:CgLOX4的cDNA全长为1 862 bp,开放阅读框为834 bp,可编码277个氨基酸,等电点为8.36,预测其氨基酸序列只含有一个保守的HOX结构域;CgLOX4在长牡蛎成体各组织中均有表达,其中在闭壳肌中的表达量显著高于肝胰腺、鳃、血淋巴、性腺和唇瓣组织(P<0.05);CgLOX4在所检测的早期胚胎中均有表达,其中在受精卵和4细胞期的表达量相对较高;CgLOX4阳性信号主要分布在担轮幼虫壳形成区、D型幼虫内脏团和壳顶幼虫消化腺中。研究表明,CgLOX4是长牡蛎早期发育过程中较早激活的转录因子之一,可能在调控幼虫壳形成和消化器官形成等方面发挥重要作用。

Abstract

To analyze the expression characteristics of CgLOX4 during larval development, the full-length cDNA sequence of LOX4 was amplified from Pacific oyster (Crassostrea gigas) by using PCR, and the expression characteristics of CgLOX4 in different stages of larval development were further analyzed by qRT-PCR and whole-mount immunofluorescence experiments. It was found that the CgLOX4 gene had full-length cDNA sequence of 1 862 bp with an open reading frame of 834 bp encoding 277 amino acids, with the isoelectric point of 8.36. The amino acid sequence of CgLOX4 gene contained a conserved HOX domain, and CgLOX4 gene was expressed in all the tested tissues of adult oyster, with significantly higher expression level in the adductor muscle than that in hepatopancreas, gills, hemolymph, gonad and labial palps tissue (P<0.05). CgLOX4 was expressed in all the examined early embryos, with relatively high expression level in fertilized eggs, and four-cells. The whole-mount immunofluorescence assay revealed that the positive signals of LOX4 were mainly distributed in the shell field of trochophore, the visceral mass of D-shape larvae and the digestive gland of umbo larvae. The findings indicate that CgLOX4 gene is one of the transcription factors activated earlier in the early development process of the oyster, and may play an important role in regulating the formation of larval shell and digestive organ.

关键词

长牡蛎 / LOX4 / 幼虫发育 / 分布特征

Key words

Crassostrea gigas /  LOX4 / larval development / distribution characteristics

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导出引用
胡悦, 冷金源, 孙洁洁, 王玲玲, 宋林生. CgLOX4基因在长牡蛎幼虫发育中的表达特征[J]. 大连海洋大学学报, 2024, 39(1): 38-47 https://doi.org/10.16535/j.cnki.dlhyxb.2023-120
HU Yue, LENG Jinyuan, SUN Jiejie, WANG Lingling, SONG Linsheng. Expression characteristics of CgLOX4 gene in larval development of Pacific oyster (Crassostrea gigas)[J]. Journal of Dalian Fisheries University, 2024, 39(1): 38-47 https://doi.org/10.16535/j.cnki.dlhyxb.2023-120
中图分类号: S 917.4   

基金

国家自然科学基金(32222086,32173002);大连市优秀青年科技人才项目(2022RY01)

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