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| 绵羊繁殖轴相关组织TPH1和TPH2基因表达分析及TPH2基因多态性与产羔数的关系 |
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李华振1,2,狄冉1,郭晓飞3,张金龙3,张效生3,马琳1,刘武军2*,储明星1*
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| (1.中国农业科学院 北京畜牧兽医研究所/农业部动物遗传育种与繁殖重点实验室, 北京 100193;2.新疆农业大学 动物科学学院, 乌鲁木齐 830052;3.天津市畜牧兽医研究所, 天津 300381) |
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| 摘要: |
| 为探讨TPH1和TPH2基因在绵羊不同繁殖状态下的表达差异及TPH2基因多态性与小尾寒羊产羔数的关系，利用实时荧光定量PCR技术检测该2个基因在不同繁殖状态下成年苏尼特羊(短光照3只，长光照3只)和小尾寒羊(卵泡期3只，黄体期3只)的大脑、小脑、下丘脑、松果体、垂体、卵巢、输卵管、子宫、肾脏和肾上腺等10种组织中的相对表达量；同时采用Sequenom MassARRAY®；SNP技术检测TPH2基因单核苷酸多态位点SNP在常年发情绵羊品种(小尾寒羊、湖羊和策勒黑羊)和季节性发情绵羊品种(滩羊、苏尼特羊和草原型藏羊)的多态性，并将基因多态性与小尾寒羊产羔数进行关联分析。结果表明:该2 个基因在2 个绵羊品种各组织广泛表达，TPH1基因在苏尼特羊下丘脑和松果体中短光照表达量极显著高于长光照(P<0.01)，在苏尼特羊垂体中短光照表达量显著高于长光照(P<0.05)，在小尾寒羊垂体和卵巢中卵泡期显著低于黄体期(P<0.05)；TPH2基因在苏尼特羊卵巢组织长光照下的表达量显著高于短光照(P<0.05)，TPH2基因在小尾寒羊下丘脑组织中卵泡期的表达量极显著高于黄体期(P<0.01)。小尾寒羊中TPH2基因g.107854166C>T和g.1078541669C>T 2个SNP位点关联分析表明，该2 个位点与季节性发情性状和小尾寒羊第1、2以及第3胎产羔数均无显著关联(P>0.05)。综上，TPH1和TPH2基因在苏尼特羊绵羊下丘脑和松果体中呈季节性光照节律差异表达，暗示其可能参与季节性发情上游基因的调控，而TPH2基因可能参与调控小尾寒羊下丘脑中激素分泌和卵巢发育，但g.107854166C>T和g.1078541669C>T不是调控季节性发情和小尾寒羊产羔数的关键位点。 |
| 关键词: 绵羊 TPH基因 性腺轴 季节性繁殖 基因表达 |
| DOI：10.11841/j.issn.1007-4333.2020.03.05 |
| 投稿时间：2019-08-20 |
| 基金项目:国家自然科学基金项目(31772580)；国家肉羊产业技术体系专项(CARS-38)；中国农业科学院科技创新工程(ASTIP-IAS13,CAAS-XTCX2016010-01-03,CAAS-XTCX2016010-03-03,CAAS-XTCX2016011-02-02)；农业科研杰出人才及其创新团队项目(农办人［2015］62号)；国家万人计划科技创新领军人才项目(W02020274)；天津市农业科技成果转化与推广项目(201704020) |
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| Expression analysis of TPH1 and TPH2 genes in sheep reproductive axis and the association between TPH2 gene polymorphism and litter size |
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LI Huazhen1,2,DI Ran1,GUO Xiaofei3,ZHANG Jinlong3,ZHANG Xiaosheng3,MA Lin1,LIU Wujun2*,CHU Mingxing1*
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| (1.Institute of Animal Science/Key Laboratory of Animal Genetics, Breeding and Reproduction of Ministry of Agriculture and Rural Affairs, Chinese Academy of Agricultural Sciences, Beijing 100193, China;2.College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China;3.Tianjin Institute of Animal Sciences, Tianjin 300381, China) |
| Abstract: |
| In order to investigate the expression patterns of TPH1 and TPH2 genes in different reproductive states of sheep and the relationship between TPH2 gene polymorphism and litter size, real-time quantitative PCR was used to detect their relative expression levels in 10 types of tissues including brain, cerebellum, hypothalamus, pineal gland, pituitary gland, ovary, fallopian tube, uterus, kidney and adrenal gland of Sunite sheep(3 in short day/3 in long day)and Small Tail Han sheep(3 in the follicular phase/3 in the luteal phase); Sequenom MassARRAY® SNP technique was used to detect single-nucleotide polymorphic SNPs of TPH2 gene in perennial estrus sheep breeds(Small Tail Han sheep, Hu sheep and Cele black sheep)and seasonal estrus sheep breeds(Tan sheep, Sunite sheep and Tibetan sheep)and the correlations between gene polymorphism and Small Tail Han sheep litter size was analyzed. The results showed that: TPH1 gene expression in hypothalamus and pineal gland of Sunite sheep was extremely higher in short day than that in long day(P<0. 01). The TPH1 gene expression of Sunite sheep in pituitary was significantly higher(P<0. 05)in short day than that in long day, and the TPH1 gene expression of Small Tail Han sheep follicular phase in pituitary and ovary was significantly lower than that in luteal phase(P<0. 05). The expression of TPH2 gene was higher in pineal gland and kidney of Sunite sheep. The TPH2 gene expression of ovarian tissue of Sunite sheep in long day was significantly higher than that in short day(P<0. 05). For Small Tail Han sheep, the TPH2 gene expression of follicular phase in hypothalamic tissue was extremely higher than that in luteal phase(P<0. 01). Correlation analysis between the two SNP loci and the litter size of TPH2 gene g. 107854166C>T and g. 1078541669C>T in Small Tail Han sheep showed that there was no significant correlation between the two loci and the number of litter size and seasonal estrus in Small Tail Han sheep 1, 2 and 3(P>0. 05). TPH1 and TPH2 are differentially expressed by seasonal light rhythm, suggesting that they are involved in the regulation of upstream genes in seasonal estrus. TPH2 gene may be involved in the regulation of hormone secretion and ovarian development in the hypothalamus of Small Tail Han sheep, but g. 107854166C>T and g. 1078541669C>T are not key sites affecting the number of Small Tail Han sheep litter size and seasonal estrus. |
| Key words: sheep TPH HPGA seasonal reproduction gene expression |
