研究论文 RESEARCH PAPER

妊娠后期饲粮蛋白质水平对藏母羊泌乳性能的影响及乳腺转录组分析

  • 周力 ,
  • 姜洋洋 ,
  • 李蒋伟 ,
  • 张峰硕 ,
  • 杨葆春 ,
  • 王志有 ,
  • 桂林生 ,
  • 侯生珍
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  • 青海大学农牧学院, 西宁 810016
周力(1994-),男,安徽定远人,博士研究生,从事动物生物技术及遗传育种研究。E-mail:472737885@qq.com

收稿日期: 2021-10-27

  网络出版日期: 2022-07-14

基金资助

青海省农业农村厅农牧业科技创新平台(QNKP20210602);青海省林草局青海湖-祁连山保护体系(K992138);青海省乐都区基层农技推广体系改革(K151942);青海省贵南县黑藏羊高效繁育基地与产业化服务(2020GN118)

Effects of Dietary Protein Level during Late Gestation on Lactation Performance of Tibetan Ewes and Analysis of Mammary Gland Transcriptome

  • ZHOU Li ,
  • JIANG Yangyang ,
  • LI Jiangwei ,
  • ZHANG Fengshuo ,
  • YANG Baochun ,
  • WANG Zhiyou ,
  • GUI Linsheng ,
  • HOU Shengzhen
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  • College of Agriculture and Animal Husbandry, Qinghai University, Xining 810016, China

Received date: 2021-10-27

  Online published: 2022-07-14

摘要

本试验旨在探究妊娠后期饲粮蛋白质水平对藏母羊产奶量、乳成分及乳腺组织基因表达的影响。选取妊娠后期藏母羊60只,随机分成2组,每组30只,分别饲喂蛋白质水平为7.5%(LP组)和11.5%(HP组)的饲粮。试验期65 d(配种后第85~150天),其中预试期5 d,正试期60 d。试验结束后,通过RNA-seq技术进行乳腺转录组测序,并对差异表达基因(DEGs)进行GO功能注释、KEGG富集分析和实时荧光定量PCR(RT-qPCR)验证。结果显示:HP组羔羊的平均日增重显著高于LP组(P<0.05)。相较于LP组,HP组藏母羊的日产奶量、乳糖率、乳脂率以及乳蛋白率显著提高(P<0.05)。乳腺转录组分析显示,LP组和HP组分别产生了152 766 266和159 469 210个高质量reads,共筛选到40个DEGs,其中11个上调基因,29个下调基因;GO富集分类显示,DEGs被注释到29个条目,主要集中在细胞进程、细胞组分及结合等功能;KEGG通路分析表明,DEGs富集到30条差异信号通路,主要包括Rap1信号通路、烟酸和烟酰胺代谢、果糖和甘露糖代谢、矿物质吸收、磷脂酰肌醇-3-激酶-丝/苏氨酸蛋白激酶信号通路;通过RT-qPCR技术对2组藏母羊乳腺组织中胰岛素样生长因子1(IGF1)、细胞周期蛋白1(CCND1)、信号转导和转录激活因子3(STAT3)、蛋白络氨酸激酶(SRC)和Ⅵ型胶原蛋白A5(COL6A5)等5个调控糖、脂代谢的DEGs进行表达验证,所得结果与RNA-seq测序结果相一致。综上可知,提高妊娠后期藏母羊饲粮蛋白质水平不仅可提高羔羊的平均日增重,还能调控藏母羊乳腺糖、脂代谢,改善其泌乳性能。

本文引用格式

周力 , 姜洋洋 , 李蒋伟 , 张峰硕 , 杨葆春 , 王志有 , 桂林生 , 侯生珍 . 妊娠后期饲粮蛋白质水平对藏母羊泌乳性能的影响及乳腺转录组分析[J]. 动物营养学报, 2022 , 34(7) : 4610 -4620 . DOI: 10.3969/j.issn.1006-267x.2022.07.049

Abstract

This study investigated the effects of dietary protein level during late gestation period on milk yield, milk composition and gene expression of mammary tissue in Tibetan ewes. Sixty late-gestation ewes were randomly divided into 2 groups with 30 ewes per group. Ewes in the 2 groups were fed diets with 7.5% (LP group) and 11.5% (HP group) protein, respectively. The experiment lasted for 65 days (days 85 to 150 after mating) with 5 days for adaptation and 60 days for test. After the experiment, RNA-seq technology was employed for mammary tissue. GO functional annotation and KEGG enrichment analysis were used to determine the functions of differentially expressed genes (DEGs), and quantitative real-time PCR (RT-qPCR) was employed for the verification of DEGs. The results showed as follows:the average daily gain of newborn lambs in HP group were significantly higher than that in LP group (P<0.05). Compared with LP group, the daily milk yield, milk lactose percentage, milk fat percentage and milk protein percentage of Tibetan ewes in HP group were significantly increased (P<0.05). Transcriptome analysis of mammary gland showed that 152 766 266 and 159 469 210 clean reads were produced in LP and HP groups, respectively. A total of 40 DEGs were selected, among which 11 genes were up-regulated and 29 genes were down-regulated. The functions of these DEGs were classified into 29 GO items, and the DEGs were mainly concentrated in cellular process, cell part, binding and other functions. KEGG pathway analysis showed that 30 differentially signaling pathways were enriched (P<0.05), including Rap1 signaling pathway, nicotinate and nicotinamide metabolism, fructose and mannose metabolism, mineral absorption and phosphatidylinositol-3-kinase/serine-threonine kinase (PI3K-Akt) signaling pathway. Using the RT-qPCR technology, the expression of glucose and lipid related DEGs, such as insulin-like growth factor 1 (IGF1), cyclin D1 (CCND1), signal transduction and transcriptional activation factor 3 (STAT3), sarcoma (SRC) and type Ⅳ collagen A5 (COL6A5) were verified, which confirmed the reliability of RNA-seq results. In conclusion, increasing dietary protein level of Tibetan ewes during late gestation can not only improve the average daily gain of lambs, but also regulate the glucose and lipid metabolism of Tibetan ewes and affect their lactation performance.

参考文献

[1] LI H,WANG W,ZHANG L,et al.Identification of a long noncoding RNA-associated competing endogenous RNA network in intracranial aneurysm[J].World Neurosurgery,2017,97:684-692.e4.
[2] CHEN J,XU J,LI Y S,et al.Competing endogenous RNA network analysis identifies critical genes among the different breast cancer subtypes[J].Oncotarget,2017,8(6):10171-10184.  
[3] LEMAY D G,BALLARD O A,HUGHES M A,et al.RNA sequencing of the human milk fat layer transcriptome reveals distinct gene expression profiles at three stages of lactation[J].PLoS One,2013,8(7):e67531.
[4] CUI X G,HOU Y L,YANG S H,et al.Transcriptional profiling of mammary gland in Holstein cows with extremely different milk protein and fat percentage using RNA sequencing[J].BMC Genomics,2014,15(1):226.
[5] MOBUCHON L,MARTHEY S,BOUSSAHA M,et al.Annotation of the goat genome using next generation sequencing of microRNA expressed by the lactating mammary gland:comparison of three approaches[J].BMC Genomics,2015,16(1):285.
[6] 李海琴,贾建磊,李鹏翔,等.饲粮不同营养水平对小尾寒羊泌乳性能及生殖激素的影响[J].青海大学学报(自然科学版),2019,37(4):49-54. LI H Q,JIA J L,LI P X,et al.Effects of different dietary nutrition levels on lactation performance and reproductive hormone of small tailed Han sheep[J].Journal of Qinghai University(Natural Science),2019,37(4):49-54.(in Chinese)
[7] 许会芬,罗军,朱越,等.日粮中不同能量和蛋白水平对西农萨能羊泌乳量、乳成分及血浆生化指标的影响[J].黑龙江畜牧兽医,2013(1):1-4. XU H F,LUO J,ZHU Y,et al.Effects of different energy and protein levels in the diets on the amount of lactation,milk composition and plasma biochemical indices of Xinong Saanen dairy goats[J].Heilongjiang Animal Science and Veterinary Medicine,2013(1):1-4.(in Chinese)
[8] 周力,李蒋伟,张莹莹,等.冬季补饲对妊娠后期藏母羊瘤胃挥发性脂肪酸、微生物群落结构和羔羊发育的影响[J].安徽农业大学学报,2021,48(4):600-606. ZHOU L,LI J W,ZHANG Y Y,et al.Effects of winter supplementary feeding on rumen volatile fatty acids,microbial community structure and lamb development of Tibetan ewe in late pregnancy[J].Journal of Anhui Agricultural University,2021,48(4):600-606.(in Chinese)
[9] 王继卿,郝志云,沈继源,等.小尾寒羊泌乳性状重要lncRNAs的筛选、鉴定及功能分析[J].中国农业科学,2021,54(14):3113-3123. WANG J Q,HAO Z Y,SHEN J Y,et al.Screening,identification and functional analysis of important LncRNAs for lactation traits in small-tailed Han sheep[J].Scientia Agricultura Sinica,2021,54(14):3113-3123.(in Chinese)
[10] LI Y X,FENG X P,WANG H L,et al.Transcriptome analysis reveals corresponding genes and key pathways involved in heat stress in Hu sheep[J].Cell Stress and Chaperones,2019,24(6):1045-1054.  
[11] 王彩莲,吴建平,刘立山,等.精料补饲水平对藏羊屠宰性能和器官发育的影响[J].草业科学,2021,38(2):348-357. WANG C L,WU J P,LIU L S,et al.Effects of concentrate supplementation amount on slaughter performance and organ development of Tibetan sheep[J].Pratacultural Science,2021,38(2):348-357.(in Chinese)
[12] GRIFFITHS K J,RIDLER A L,HEUER C,et al.The effect of liveweight and body condition score on the ability of ewe lambs to successfully rear their offspring[J].Small Ruminant Research,2016,145:130-135.
[13] 周爱民,俄木曲者,王育伟,等.补饲不同蛋白水平日粮对北川白山羊生产性能和屠宰性能的影响[J].家畜生态学报,2020,41(2):29-33,39. ZHOU A M,EMU Q Z,WANG Y W,et al.Effects of different dietary protein levels on production and slaughter performance of Beichuan white goat[J].Acta Ecologae Animalis Domastici,2020,41(2):29-33,39.(in Chinese)
[14] 雷耀庚,李碧波,王兴涛,等.日粮蛋白水平对陕北白绒山羊日增重、养分消化和精液品质的影响[J].畜牧与兽医,2015,47(11):22-27. LEI Y G,LI B B,WANG X T,et al.Effects of dietary protein levels on average daily gain,nutrient degradation and semen quality in Shanbei white cashmere goat bucks[J].Animal Husbandry & Veterinary Medicine,2015,47(11):22-27.(in Chinese)
[15] ARTHINGTON J D,KALMBACHER R S.Effect of early weaning on the performance of three-year-old,first-calf beef heifers and calves reared in the subtropics[J].Journal of Animal Science,2003,81(5):1136-1141.  
[16] 王思宇,赵小琪,李银江,等.日粮钙水平对云南半细毛羊乳成分的影响[J].中国饲料,2021(13):61-65. WANG S Y,ZHAO X Q,LI Y J,et al.Effect of dietary calcium on milk composition of Yunnan semi-fine-wool sheep[J].China Feed,2021(13):61-65.(in Chinese)
[17] 于子洋,袁翠林,王利华,等.蛋白质水平对崂山奶山羊泌乳性能的影响[J].中国畜牧杂志,2015,51(7):32-36. YU Z Y,YUAN C L,WANG L H,et al.Effects of dietary protein level on lactation performance of laoshan dairy goat[J].Chinese Journal of Animal Science,2015,51(7):32-36.(in Chinese)
[18] AMIRABADI FARAHANI T,AMANLOU H,FARSUNI N E,et al.Interactions of protein levels fed to Holstein cows pre- and postpartum on productive and metabolic responses[J].Journal of Dairy Science,2019,102(1):246-259.  
[19] 田平,孙利利,董海波,等.饲喂高精料日粮对泌乳期山羊泌乳性能及抗氧化能力的影响[J].草业学报,2017,26(4):99-105. TIAN P,SUN L L,DONG H B,et al.Effects of feeding high concentrate diets on lactation indicators and antioxidant levels in the mammary glands of lactating goats[J].Acta Prataculturae Sinica,2017,26(4):99-105.(in Chinese)
[20] 吴敏,罗军,姚大为,等.日粮中不同蛋白质和L-赖氨酸水平对西农萨能羊泌乳性能和血浆生化指标的影响[J].中国畜牧杂志,2013,49(9):34-38. WU M,LUO J,YAO D W,et al.Effects of different dietary protein and L-lysine levels on lactation performance and plasma biochemical indices of Xinong Saanen sheep[J].Chinese Journal of Animal Science,2013,49(9):34-38.(in Chinese)
[21] 李歆.日粮中不同能量和蛋白水平对西农萨能奶山羊泌乳性能及乳成分的影响[D].硕士学位论文.杨凌:西北农林科技大学,2012. LI X.Effect of different diet energy and protein level on milk performance and composition in Xinong Saanen dairy goats[D].Master's Thesis.Yangling:Northwest A&F University,2012.(in Chinese)
[22] WHELAN S J,PIERCE K M,MCCARNEY C,et al.Effect of supplementary concentrate type on nitrogen partitioning in early lactation dairy cows offered perennial ryegrass-based pasture[J].Journal of Dairy Science,2012,95(8):4468-4477.  
[23] LLOYD-LEWIS B.Multidimensional imaging of mammary gland development:a window into breast form and function[J].Frontiers in Cell and Developmental Biology,2020,8:203.
[24] FARMER C.Review:mammary development in swine:effects of hormonal status,nutrition and management[J].Canadian Journal of Animal Science,2013,93(1):1-7.  
[25] HUE-BEAUVAIS C,FAULCONNIER Y,CHARLIER M,et al.Nutritional regulation of mammary gland development and milk synthesis in animal models and dairy species[J].Genes,2021,12(4):523.
[26] REZAEI R,WU Z L,HOU Y Q,et al.Amino acids and mammary gland development:nutritional implications for milk production and neonatal growth[J].Journal of Animal Science and Biotechnology,2016,7:20.
[27] GÓMEZ-MARTÍNEZ J E,RODRÍGUEZ-OSORIO N,ECHEVERRI-ZULUAGA J J,et al.The energy level of the diet in Holstein cows in the first third of lactation alters the expression of immune genes in the mammary gland[J].Journal of Animal Physiology and Animal Nutrition,2021,105(2):219-231.  
[28] GRAJALES S M B,ZULUAGA J J E,HERRERA A L,et al.RNA-seq differential gene expression analysis in mammary tissue from lactating dairy cows supplemented with sunflower oil[J].Animal Production Science,2020,60(6):758-771.  
[29] CONTE G,GIORDANI T,VANGELISTI A,et al.Transcriptome adaptation of the ovine mammary gland to dietary supplementation of extruded linseed[J].Animals,2021,11(9):2707.
[30] AKERS R M.Major advances associated with hormone and growth factor regulation of mammary growth and lactation in dairy cows[J].Journal of Dairy Science,2006,89(4):1222-1234.  
[31] 高琴,隋啸一,李福昌,等.不同蛋白水平饲粮对肉兔骨骼肌发育及相关因子表达的影响[J].中国兽医学报,2016,36(9):1613-1618. GAO Q,SUI X Y,LI F C,et al. Effect of dietary protein on muscle development and relative factors of rabbit[J].Chinese Journal of Veterinary Science,2016,36(9):1613-1618.(in Chinese)
[32] QIE S,DIEHL J A.Cyclin D1,cancer progression,and opportunities in cancer treatment[J].Journal of Molecular Medicine,2016,94(12):1313-1326.  
[33] 李冬辉.奶牛泌乳功能基因研究[J].牡丹江大学学报,2010,19(10):116-117,125. LI D H.Study on milk function genes of dairy cows[J].Journal of Mudanjiang University,2010,19(10):116-117,125.(in Chinese)
[34] WATKIN H,RICHERT M M,LEWIS A,et al.Lactation failure in Src knockout mice is due to impaired secretory activation[J].BMC Developmental Biology,2008,8:6.
[35] 刘文艳,鞠志花,刘顺德,等.Src基因研究进展[J].家畜生态学报,2010,31(2):87-90. LIU W Y,JU Z H,LIU S D,et al.Progress in the study on Src gene[J].Acta Ecologae Animalis Domastici,2010,31(2):87-90.(in Chinese)
[36] 孙立峰.脂代谢相关分子Col6a5和Gpr1在高雄激素化小鼠中的作用及其机理[D].博士学位论文.北京:中国科学院大学,2019. SUN L F.Effect of lipid metabolism related molecules Col6a5 and Gpr1 on hyperandrogenic mice and its mechanism[D].Ph.D.Thesis.Beijing:University of Chinese Academy of Sciences,2019.(in Chinese)
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