种公鸡精清miRNA调控鸡蛋营养积累的分子作用初探  

Primary Exploration on Molecular Mechanisms of Breeder Cocks’Spermic miRNA Modulating Nutrient Accumulation of Eggs

作　　者：刘喆 樊晓钰 王佳媚 张晓叶 李玉龙 LIU Zhe;FAN Xiaoyu;WANG Jiamei;ZHANG Xiaoye;LI Yulong(College of Basic Medical Science,Shaanxi University of Chinese Medicine,Xianyang 712046,China)

机构地区：[1]陕西中医药大学基础医学院,咸阳712046

出　　处：《动物营养学报》2025年第4期2684-2696,共13页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：略阳乌鸡产业发展研究院(WJYJY-2021-10);秦创原中医药产业创新聚集区项目(中医药大学、医疗机构)(L2024-QCY-ZYYJJQ-X214);陕西省科技厅青年项目(2023-JC-QN-0266)。

摘　　要：为探究种公鸡精清miRNA对母鸡繁殖过程营养分配的影响,选取体重接近的健康略阳乌鸡种公鸡40只、母鸡120只作为试验动物,种公鸡随机分为自由采食组和限饲组(饲喂自由采食量的70%),每组4个重复,每个重复5只鸡,每个重复随机配套15只母鸡。采用高通量测序分析限饲对种公鸡精清miRNA的影响,结合配套母鸡卵巢上皮转录组测序(RNA-seq)进行关联分析,筛选精清miRNA中调控母鸡卵巢上皮营养代谢和转运的关键miRNA及其靶基因,并结合蛋黄代谢组确定种公鸡营养干预对母鸡繁殖过程营养分配的调控效应。结果表明:限饲可显著改变种公鸡精清中52种miRNA的表达水平,精清miRNA测序与卵巢上皮转录组测序关联分析显示限饲种公鸡可以gga-let-7b、gga-miR-200a-3p、gga-miR-2954等高表达miRNA为媒介序列特异性调控其配套母鸡卵巢上皮中内吞和自噬信号通路调控因子[如高迁移率族蛋白1(HMGB1)、Ras相关蛋白10(RAB10)、F肌动蛋白加帽蛋白α2亚基(CAPZA2)和翻译调节肿瘤蛋白1(TPT1)等]的表达。内吞和自噬信号通路均与营养物质代谢和转运有关,蛋黄代谢组进一步说明种公鸡限饲可通过调控卵巢上皮基因表达间接改变鸡蛋蛋黄中多种氨基酸、脂肪酸及其衍生物的含量,进而影响其氨基酸和能量代谢功能。总之,限饲种公鸡可以精清miRNA为媒介调控母鸡卵巢上皮内吞和自噬信号通路活性,间接影响鸡蛋蛋黄营养组成。The purpose of this study is to investigate the effects of breeder cocks’spermic miRNA on the nutrition distribution during the reproductive process of hens.A total of 40 healthy Lueyang black-bone breeder cocks and 120 hens were selected as experimental animals.The breeder cocks were randomly assigned to two groups:an ad libitum feeding group and a feed-restricted group(70%of ad libitum feed intake),with 4 replicates per group and 5 cocks per replicate.Each replicate of cocks was randomly paired with 15 hens.High-throughput sequencing was employed to investigate the effects of feed restriction on seminal miRNA of breeding cocks,and it was integrated with RNA-seq of ovarian epithelium from paired hens to perform correlation analysis,aiming to identify key miRNA in seminal fluid that regulate nutrient metabolism and transport in the ovarian epithelium,along with their target genes.Furthermore,yolk metabolomics was utilized to elucidate the impact of breeder cocks’nutritional intervention on nutrient distribution during reproduction of hens.The results showed that restricted feeding could significantly change the expression levels of 52 spermic miRNA.Association analysis of miRNA-seq and RNA-seq indicated that feed-restricted breeder cocks could specifically regulate the expression of regulatory factors in the endocytosis and autophagy signaling pathways in the ovarian epithelium of their mating hens,such as high mobility group box 1 protein(HMGB1),Ras-associated protein 10(RAB10),F-actin capping protein alpha 2 subunit(CAPZA2)and translationally regulated tumor protein 1(TPT1),through high expression of miRNA(such as gga-let-7b,gga-miR-200a-3p and gga-miR-2954)as a mediator sequence.Both endocytosis and autophagy signaling pathways were related to nutrient metabolism and transport.The yolk metabolome further indicated that feed-restricted breeder cocks could indirectly change the contents of various amino acids and fatty acids and their derivatives in egg yolk by regulating the expression of ovarian epithelial

关 键 词：略阳乌鸡 精清miRNA 内吞信号通路 自噬信号通路 鸡蛋营养组成 

分 类 号：S831[农业科学—畜牧学]