低频声波频率下瘤背石磺CaM-like、CaMKII基因的表达及功能
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上海海洋大学水产种质资源发掘与利用教育部重点实验室;水产科学国家级实验教学示范中心;海洋动物系统分类与进化上海高校重点实验室, 上海 201306

作者简介:

梁威(1993-),男,硕士研究生,研究方向为水产种质资源与苗种工程.E-mail:lw1299199029@163.com

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S917

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国家自然科学基金项目(41276157);上海高校水产学一流学科建设项目.


Expression of CaM-like and CaMKII genes in Onchidium reevesii under low frequency sound wave stimulation
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Key Laboratory of Exploration and Utilization of Aquatic Germplasm Resources of Shanghai Ocean University, Ministry of Education;National Experimental Teaching Demonstration Center of Fisheries Science;Marine Animal System Classification and Evolution Shanghai University Key Laboratory, Shanghai 201306, China

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    摘要:

    长期栖息于潮间带的动物能够感知当地的潮汐规律,形成潮汐记忆的能力。本研究以栖息于潮间带的瘤背石磺()为对象,研究其通过感知潮汐来临时产生的低频声音来调节其行为与潮汐节律相协调的分子机制。通过RACE-PCR技术克隆到基因的cDNA全长2321 bp,包括5'非编码区(UTR)366 bp,3'非编码区(UTR)1337 bp,618 bp的开放阅读框编码206个氨基酸;预测该基因编码的多肽链的原子数量是3165,分子量约为23029.64 kD,理论等电点4.64,分子式是C1018H1544N274O320S9。N端信号肽由29个氨基酸组成。氨基酸序列比对构建系统进化树,结果显示瘤背石磺Biomphalaria glabrata)的CaM-like基因在不同组织的分布情况,结果显示<0.05),推测其可能参与神经系统的可塑性调节。荧光定量结果显示基因分别在25 Hz和50 Hz低频声波下刺激12.4 h表达量较高,初步推测其能感知25~50 Hz声波频率。这可能与栖息于潮间带中的瘤背石磺长期感知12.4 h半日潮潮汐周期节律,形成潮汐记忆有关。本研究将为进一步深入了解瘤背石磺感知潮汐节律的分子机制奠定基础,也为探究海洋动物由海洋向陆地进化过程中对环境的适应机制提供依据。

    Abstract:

    Animals that inhabit intertidal areas long-term can sense the local tidal rhythm, forming a tidal memory. However, to date, no studies have investigated the mechanism of tidal rhythm sensing in in the laboratory using low-frequency tide sounds. The cDNA sequence of the gene was cloned by RACE-PCR, after which bioinformatics analysis and qRT-PCR experiments were performed. The results showed that the full-length cDNA of the gene was 2321 bp (366 bp 5' non-coding region (UTR), 1337 bp 3' non-coding region (UTR), and 618 bp open reading frame (ORF), which encoded a total 206 amino acids. The gene that encodes polypeptide chains was predicted with an atomic number of 3165, a molecular weight of approximately 23029.64 kD, a theoretical isoelectric point of 4.64, a molecular formula of C1018H1544N274O320S9, and an N-terminal signal peptide composed of 29 amino acids. The phylogenetic tree was constructed via amino acid sequence alignment. The results suggest that the like gene of , which is consistent with traditional morphological classification. The distribution of genes in different tissues was detected using qRT-PCR. The results indicated that , but the expression of ganglion was significantly higher than dorsal skin, pleopod, intestines, liver, mouthparts, and albumen gland (<0.05). We speculated that it may be involved in the plasticity regulation of the ganglion system. The quantitative fluorescence results indicated that the genes were highly expressed, underlying the stimulation of low frequency sound waves at 25 Hz and 50 Hz for 12.4 h, respectively, for which it was presumed that could sense the 25-50 Hz sound frequency. This may be related to the tidal memory formed by the long-term immersion in the 12.4 h half-day tidal cycle rhythm in the intertidal zone. This study will lay the foundation for further understanding of the molecular mechanisms of sensing tidal rhythm, and provide a basis for exploring the environmental adaptation mechanism of marine animals from ocean to land evolution.

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梁威,吴容宇,严彩瑞,杨铁柱,顾冰宁,沈和定.低频声波频率下瘤背石磺CaM-like、CaMKII基因的表达及功能[J].中国水产科学,2020,27(2):166-176
LIANG Wei, WU Rongyu, YAN Cairui, YANG Tiezhu, GU Bingning, SHEN Heding. Expression of CaM-like and CaMKII genes in Onchidium reevesii under low frequency sound wave stimulation[J]. Journal of Fishery Sciences of China,2020,27(2):166-176

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  • 在线发布日期: 2020-02-13
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