厚壳贻贝mTOR信号通路关键基因在幼虫生长发育中的表达模式分析

梁鑫杰; 徐敏慧; 叶莹莹; 李继姬

doi:10.12284/hyxb2024088

厚壳贻贝mTOR信号通路关键基因在幼虫生长发育中的表达模式分析

doi: 10.12284/hyxb2024088

浙江海洋大学国家海洋设施养殖工程技术研究中心，浙江舟山 316022

基金项目: 舟山市科技计划项目（2021C21017）。

详细信息

作者简介:
梁鑫杰（1998—），男，四川省遂宁市人，硕士研究生，主要研究方向为海洋生物学。E-mail：liangxinjie@zjou.edu.cn

通讯作者:
李继姬（1986—），男，安徽省安庆市人，博士，教授，主要研究方向为海洋生物学。E-mail：lijiji@zjou.edu.cn

中图分类号: S944.3
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出版历程
- 收稿日期: 2024-01-28
- 修回日期: 2024-04-18
- 网络出版日期: 2024-08-13
- 刊出日期: 2024-07-01

Analysis of expression patterns of key genes in the mTOR signaling pathway in the larval growth and development of Mytilus coruscus

National Engineering Research Center For Marine Aquaculture, Zhejiang Ocean University zhoushan 316022, China

摘要

摘要: 在水产养殖领域，对厚壳贻贝（Mytilus coruscus）的生长发育机制进行深入探讨，可为实现生长发育分子调控奠定理论基础。为了探究厚壳贻贝幼虫生长与发育差异基因的表达模式，本研究采用转录组测序和实时荧光定量PCR分析技术，对与生长特征相关的基因和分子途径的表达差异进行了初步研究。研究重点关注哺乳动物雷帕霉素靶蛋白（mammalian Target Of Rapamycin，mTOR）信号通路在厚壳贻贝幼虫生长和发育中的影响及调节作用。通过对不同发育时期（担轮幼虫期、D形幼虫期、壳顶幼虫期、眼点幼虫期和稚贝期）的基因表达模式进行分析，结果表明mTOR信号通路可能在厚壳贻贝幼虫的生长发育中扮演一定角色，并成功鉴定出7个与生长发育相关的关键基因。随着发育过程的推进，mTOR信号通路基因在不同发育时期的表达呈现出动态变化。其中，PI3K、TSC1/2和mTOR基因的整体表达变化趋势为先上升后下降再上升，IGFI变化趋势为先上升后下降，而EIF4B、RPS6KB和AKT基因表达则整体呈现下降趋势。这一差异化的基因表达模式反映了mTOR信号通路可能在厚壳贻贝幼虫不同发育时期中对细胞命运和生物学功能的调控，从而对其生长发育产生影响。因此，我们对mTOR信号通路关键基因在厚壳贻贝幼虫生长发育中的表达模式进行了初步探究。这些基因在调控厚壳贻贝幼虫的分子功能和生长特征方面具有重要作用，为深入理解海洋双壳动物的生理适应、代谢过程和生长变异提供了基础数据。
- 厚壳贻贝 /
- mTOR信号通路 /
- 基因 /
- 生长发育
Abstract: In the field of aquaculture, a comprehensive exploration of the growth and developmental mechanisms of Mytilus coruscus lays the theoretical foundation for molecular regulation of growth and development. In order to investigate the expression patterns of genes associated with the growth and development differences in M. coruscus larvae, this study employed transcriptome sequencing and real-time quantitative PCR analysis techniques, conducting a preliminary examination of the differential expression of genes and molecular pathways related to growth characteristics. The research places particular emphasis on the impact and regulatory roles of the mammalian target of rapamycin (mTOR) signaling pathway in the growth and development of M. coruscus larvae. By analyzing the gene expression patterns at different developmental stages (Trochophore larvae stage, D-veliger larvae stage, Veliconcha larvae stage, Pediveliger larvae stage, and Juvenile stage), the results suggest that the mTOR signaling pathway may play a specific role in the growth and development of M. coruscus larvae. Seven key genes associated with growth were successfully identified. As the developmental process progresses, the expression of genes in the mTOR signaling pathway exhibits dynamic changes across different developmental stages. Among these changes, the expression levels of the PI3K, TSCI/2, and mTOR genes initially increased, followed by a decrease, and subsequently rose again. In contrast, IGFI expression exhibited an initial increase followed by a decline. Meanwhile, the expression of EIF4B, RPS6KB, and AKT genes demonstrated an overall downward trend. This differential gene expression pattern reflects the potential regulatory influence of the mTOR signaling pathway on cell fate and biological functions during various developmental stages of M. coruscus larvae, thereby impacting their growth and development. Therefore, this study provides a preliminary exploration of the expression patterns of key genes in the mTOR signaling pathway during the growth and development of M. coruscus larvae. These genes play a crucial role in regulating the molecular functions and growth characteristics of M. coruscus larvae, offering foundational data for a deeper understanding of the physiological adaptation, metabolic processes, and growth variations in marine bivalves.
- Mytilus coruscus larvae /
- mTOR signaling pathway /
- gene /
- growth and development

HTML全文

图 1 mTOR通路中7个基因的实时荧光定量PCR分析的表达模式

Fig. 1 Expression patterns of seven genes in the mTOR pathway analyzed by RT-qPCR

下载: 全尺寸图片幻灯片

图 2 厚壳贻贝幼虫5个不同时期mTOR通路中基因表达水平变化热图。颜色标尺代表基因表达量（FPKM），以Z得分进行标准化，从最低的蓝色到最高的红色。FPKM是基于RNA-seq数据的基因表达的标准化估计值。FPKM值越大，基因表达水平越高

Fig. 2 The heatmap of gene expression levels in mTOR pathway at five different stages of Mytilus coruscus larvae. The color scale represents gene expression levels measured in Fragments Per Kilobase Million (FPKM), standardized using Z-scores, ranging from the lowest in blue to the highest in red. FPKM is a normalized estimate of gene expression based on RNA-seq data, where higher FPKM values correspond to more elevated gene expression levels

下载: 全尺寸图片幻灯片

图 3 KEGG分析中确定的分子通路—mTOR信号通路

Fig. 3 The molecular pathway identified through KEGG analysis—mTOR signaling pathway

下载: 全尺寸图片幻灯片

表 1 本研究中使用的引物序列

Tab. 1 Primer sequences used in this study

基因	引物序列5′～3′
TSC1/2-F	GAAGGGATACGAGGGGCTCA
TSC1/2-R	CCAGGTATGTAGGAATGGAAGGT
mTOR-F	TGCAGCAGTCTGAATCAGCA
mTOR-R	CCGATGCCATTCTTGCCATG
RPS6KB-F	GGCAACATAATGGCTCTGAAGG
RPS6KB-R	CCTGGGAATGCGACAGACTTA
AKT-F	TGGTGGGGCACAGGTGTAGT
AKT-R	CAAGGTGGTGTTATCTTCTTCTCA
PI3K-F	TTCTCGGGAGGAAGTGAATGA
PI3K-R	TAGTCGCCAGGTGATGTTGC
EIF4B-F	TGAGGGGTCAGGGTAGAGGT
EIF4B-R	GCATTTCATCAATAGACTGGGGTA
IGF1-F	CCTTTGACCTCTGCGGATGT
IGF1-R	TAAGCGGGGCATTCACACAT
EF1a-F*	CACCACGAGTCTCTCCCTGA
EF1a-R*	GCTGTCACCACAGACCATTCC
注：*为内参基因。

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