基于转录组数据挖掘蛋鸡产蛋前后骨代谢差异的关键候选基因  

Mining of Key Candidate Genes Involved in Bone Metabolism Differences at Pre-and Post-laying Stage Based on Transcriptome Data in Laying Hens

作　　者：张寅梁 张冉 王文君 王德贺 李兰会[1] 周荣艳[1] ZHANG Yinliang;ZHANG Ran;WANG Wenjun;WANG Dehe;LI Lanhui;ZHOU Rongyan(College of Animal Science and Technology,Hebei Agricultural University,Baoding 071001,China)

机构地区：[1]河北农业大学动物科技学院,保定071001

出　　处：《畜牧兽医学报》2024年第10期4455-4465,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金项目(32272922);河北省重点研发计划项目现代种业科技创新专项(22326319D);河北省鸡现代种业科技创新团队(21326303D);河北省现代农业产业技术体系建设专项资金(HBCT2024260204)。

摘　　要：旨在挖掘影响蛋鸡产蛋前后骨代谢差异的调控通路和关键候选基因,为进一步研究维持蛋鸡骨骼健康的调控机制提供理论依据。本研究选择15只15周龄和12只22周龄体重相近饲养条件相同的海兰灰蛋鸡,采集血液和胫骨,测定血浆钙、磷、雌激素、骨代谢相关生化指标,根据雌激素水平选择每组各6只的胫骨组织构建12个转录组文库并筛选差异表达基因,并对差异表达基因进行GO功能和KEGG富集分析。随机选择6个差异表达基因,利用qRT-PCR验证其相对表达量。15周龄与22周龄相比,蛋鸡血浆骨代谢标记物和雌激素水平均呈现显著变化(P<0.05)。构建的12个胫骨cDNA文库中得到40082240~186154554条有效碱基序列数,Q30值最少为92.18%,比对率在76.52%~90.55%之间。对15周龄与22周龄胫骨转录组进行比较,鉴定出1832个差异表达基因,其中914个基因下调,918个基因上调。GO功能注释发现,差异表达基因主要显著富集在蛋白加工、对内质网应激的反应和胶原原纤维组织等过程。KEGG富集分析发现,与骨代谢相关的显著富集通路有类固醇生物合成、甲状腺激素合成、TGF-β信号通路、MAPK信号通路、PI3K-Akt信号通路,筛选出18个这些通路共有的基因作为调控骨代谢的关键候选基因。本研究揭示了产蛋前后蛋鸡胫骨组织中基因表达存在差异,筛选到多个影响骨代谢的差异表达基因和通路,为进一步研究蛋鸡产蛋前后骨代谢转换机制提供理论依据。The study aimed to explore the regulatory pathways and important candidate genes that affect the differences in bone metabolism before and after egg laying in laying hens,providing a theoretical basis for further research on the regulatory mechanisms for maintaining bone health in laying hens.A total of 15 at 15-week-old and 12 at 22-week-old Hyline Grey laying hens with similar body weights and reared under identical conditions were selected.Blood and tibia samples were collected to measure plasma levels of calcium,phosphorus,estrogen,and other biochemical markers related to bone metabolism.Based on estrogen levels,tibia tissues from 6 hens within each age group were selected to construct 12 transcriptome libraries.Differentially expressed genes(DEGs)were then screened from these libraries,and GO functional annotation and KEGG pathway enrichment analyses were performed with DEGs.Additionally,6 randomly selected DEGs were validated for their relative expression levels using qRT-PCR.When comparing 15-week-old hens to 22-week-old hens,significant changes(P<0.05)were observed in plasma bone metabolism markers and estrogen levels in the laying hens.The clean reads of 40082240-186154554 were obtained from the 12 constructed tibial cDNA libraries,with a minimum Q30 value of 92.18%and a contrast rate between 76.52%and 90.55%.A total of 1832 DEGs were screened from the tibia of laying hens at 15 and 22 weeks of age.Among them,914 genes were downregu-lated and 918 genes were upregulated.GO functional annotation revealed that differentially ex-pressed genes were significantly enriched in protein processing,response to endoplasmic reticu-lum stress,and collagen fibril organization(P<0.05).KEGG enrichment analysis revealed sig-nificant enrichment pathways related to bone metabolism,including steroid biosynthesis,thyroid hormone synthesis,and TGF-βsignal pathway,MAPK signaling pathway,PI3K-Akt signaling pathway.Screening out 18 genes were shared by these pathways as key candidate genes for regu-lating bone metabolism.This st

关 键 词：蛋鸡 转录组 差异表达基因 生理阶段 胫骨 骨代谢 

分 类 号：S831.2[农业科学—畜牧学]