Zeng Bin, Tang Min, Chen Ting, Jiang Yuzhang, Tang Wei, Yu Guiyang
Yongzhou Engineering Technology Research Center for Efficient and Healthy Breeding of Livestock and Poultry, College of Agriculture, Yongzhou Vocational Technical College, Yongzhou 425000, PR China.
Yongzhou Engineering Technology Research Center for Efficient and Healthy Breeding of Livestock and Poultry, College of Agriculture, Yongzhou Vocational Technical College, Yongzhou 425000, PR China.
Poult Sci. 2025 Jun 5;104(9):105354. doi: 10.1016/j.psj.2025.105354.
Skeletal muscle development at the embryonic stage is critical for muscle growth in poultry, during which microRNAs (miRNAs) play a vital role. Geese are one of the main poultry species, but how miRNAs regulate skeletal muscle development in geese during the embryonic stage remains poorly understood. This study characterized histological features and miRNA expression profiles during leg muscle development in Daozhou Grey goose (a prized Chinese breed) embryos at three stages (E14, E21, and E28). The objective was to identify differentially expressed miRNAs (DEMs) associated with muscle development in Daozhou Grey goose and utilize bioinformatics to predict the potential biological functions of these DEMs. Histological observations revealed stage-specific muscle development, marked by myoblast proliferation at E14, differentiation at E21 and muscle fiber maturation at E28. A total of 340 known and 270 novel miRNAs were identified by miRNA sequencing, with miR-148a-3p, miR-1a-3p, miR-100-5p, miR-206, and miR-92-3p being the most abundant. Furthermore, differential expression analysis identified 105, 107 and 70 DEMs across E14 vs E21, E14 vs E28, and E21 vs E28 comparisons, respectively. The expression patterns of the twelve randomly selected DEMs (9 known and 3 novel miRNAs) were validated by Quantitative real-time polymerase chain reaction (qRT-PCR), confirming the sequencing results. Functional enrichment analysis of predicted miRNA target genes revealed involvement of the MAPK, TGF-β and Notch signaling pathways in embryonic myogenesis. The miRNA-mRNA interaction network analysis of the top 10 most DEMs and their target genes revealed key regulatory nodes such as let-7k-5p-MAP3K1, miR-133a-3p-FZD7, miR-133c-3p-STAT3, miR-187-3p-ZEB2, and miR-205b-SETD3, with inverse expression patterns validated by qRT-PCR. It was hypothesized that the above pathways as well as these miRNA-mRNA target relationships play an important role in the regulating embryonic muscle growth and development of the Daozhou Grey goose. These findings provide the first comprehensive miRNAs profile of embryonic leg muscle in vivo in geese, offering a basic reference and theoretical framework for further research on molecular regulatory mechanisms of goose muscle development.
胚胎期骨骼肌发育对家禽肌肉生长至关重要，在此过程中，微小RNA（miRNA）发挥着关键作用。鹅是主要家禽品种之一，但miRNA如何在胚胎期调控鹅的骨骼肌发育仍知之甚少。本研究对道州灰鹅（一种珍贵的中国品种）胚胎三个阶段（E14、E21和E28）腿部肌肉发育过程中的组织学特征和miRNA表达谱进行了表征。目的是鉴定与道州灰鹅肌肉发育相关的差异表达miRNA（DEM），并利用生物信息学预测这些DEM的潜在生物学功能。组织学观察揭示了特定阶段的肌肉发育，其特征为E14期成肌细胞增殖、E21期分化以及E28期肌纤维成熟。通过miRNA测序共鉴定出340个已知miRNA和270个新miRNA，其中miR-148a-3p、miR-1a-3p、miR-100-5p、miR-206和miR-92-3p含量最为丰富。此外，差异表达分析分别在E14与E21、E14与E28以及E21与E28的比较中鉴定出105个、107个和70个DEM。通过定量实时聚合酶链反应（qRT-PCR）验证了随机选择的12个DEM（9个已知miRNA和3个新miRNA）的表达模式，证实了测序结果。对预测的miRNA靶基因进行功能富集分析，揭示了丝裂原活化蛋白激酶（MAPK）、转化生长因子-β（TGF-β）和Notch信号通路参与胚胎期肌生成。对前10个差异最大的DEM及其靶基因进行的miRNA- mRNA相互作用网络分析揭示了关键调控节点，如let-7k-5p-MAP3K1、miR-133a-3p-FZD7、miR-133c-3p-STAT3、miR-187-3p-ZEB2和miR-205b-SETD3，qRT-PCR验证了它们的反向表达模式。据推测，上述信号通路以及这些miRNA-mRNA靶标关系在调控道州灰鹅胚胎期肌肉生长发育中起重要作用。这些发现首次提供了鹅胚胎期腿部肌肉在体内的全面miRNA图谱，为进一步研究鹅肌肉发育的分子调控机制提供了基础参考和理论框架。
