大黄鱼视蛋白的鉴定、进化及表达分析

Identification, evolution, and expression analysis of opsin in large yellow croaker

  • 摘要:
    目的 视蛋白(opsin)是介导鱼类光信号感受与传导的蛋白,在视觉发育、摄食行为及环境适应中发挥重要作用。为解析大黄鱼视蛋白基因家族的组成特征、进化规律及其表达模式。
    方法 本研究采用生物信息学方法鉴定大黄鱼opsin基因家族成员,分析其理化性质、保守基序、结构域特征及系统发育关系,并通过qRT-PCR技术检测其在早期发育阶段的表达模式及不同光色处理下的响应特征。
    结果 大黄鱼共鉴定出25个opsin基因,编码区长度为334–540个氨基酸,预测分子量为37.1–62.3 ku,等电点为5.95–9.57,均定位于细胞膜。所有opsin蛋白均含有7个保守基序和典型的七次跨膜结构域(7tm_1),归属于Class A型G蛋白偶联受体超家族,二级结构以α-螺旋(30.07 %–40.41 %)和无规则卷曲(44.54%–55.93%)为主。系统发育分析表明,大黄鱼opsin基因可分为视觉型和非视觉型两大类,视觉型opsin包括rhoopn1lw1opn1sw2rh2a;非视觉型opsin包括opn3opn4、opn5等多个亚型。转录组分析显示,大黄鱼opsin基因在1~35日龄发育阶段整体表达水平较低,其中rhoopn1lw1在15日龄显著上调。光谱胁迫结果表明,蓝光可显著上调rh2aopn1sw2的表达(P<0.05),提示其在视觉形成及光谱调控中发挥核心作用。
    结论 大黄鱼opsin基因家族在结构上高度保守,并在发育过程中呈现阶段性和光谱依赖性的表达特征。

     

    Abstract: Opsins are light-sensitive G protein–coupled receptors that mediate photoreception and phototransduction in fish and play essential roles in visual development, light-dependent behavior, and adaptation to aquatic environments, and in teleosts the expansion and diversification of the opsin gene family are considered key molecular mechanisms underlying spectral tuning to complex underwater light conditions; however, comprehensive characterization of opsins in the economically important large yellow croaker (Larimichthys crocea) remains limited. The present study aimed to systematically identify the opsin gene family in large yellow croaker, elucidate its structural and evolutionary features, and clarify developmental and light spectrum–dependent expression patterns. Genome-wide bioinformatics analyses were performed to identify opsin genes, followed by characterization of physicochemical properties, conserved motifs, transmembrane domains, secondary structure composition, subcellular localization, and phylogenetic relationships, while transcriptomic data and quantitative real-time PCR (qRT-PCR) were used to examine expression dynamics during early development (1–35 days post-hatching, dph) and transcriptional responses under different monochromatic light treatments. A total of 25 opsin genes were identified, encoding proteins of 334–540 amino acids with predicted molecular weights of 37.1–62.3 kDa and isoelectric points ranging from 5.95 to 9.57, all of which were predicted to localize to the cell membrane and contained seven highly conserved motifs and the canonical seven-transmembrane (7tm_1) domain characteristic of Class A G protein–coupled receptors. Secondary structure prediction revealed that α-helices (30.07%–40.41%) and random coils (44.54%–55.93%) were the predominant structural elements. Phylogenetic analysis classified these genes into visual opsins, including rho, opn1lw1, opn1sw2, and rh2a, and non-visual opsins, including opn3, opn4, opn5, and related subtypes. Transcriptome profiling showed that most opsin genes exhibited relatively low expression levels throughout early development, whereas rho and opn1lw1 displayed pronounced upregulation at 15 dph, suggesting a critical window for rapid visual system maturation. Furthermore, light spectrum experiments demonstrated that blue light exposure significantly upregulated the expression of rh2a and opn1sw2 compared with other light treatments (P < 0.05), indicating high sensitivity of these opsins to short-wavelength light and their involvement in spectral regulation. Collectively, these results demonstrate that the opsin gene family of large yellow croaker is structurally conserved yet exhibits distinct stage-specific and spectrum-dependent expression patterns, providing molecular evidence for visual development and spectral adaptation and offering practical implications for optimizing light environments, feeding strategies, and larval rearing protocols in aquaculture production.

     

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