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    摘要：探索黄体期不同营养水平及静脉注射葡萄糖通过Akt信号通路影响湖羊卵泡发育的分子机制。挑选体况相近的纯种湖羊24只，情期第8天将试验羊随机分为对照组(M)、限饲组（0.5*M）、限饲+葡萄糖注射组（0.5*M+Glu），第14天起试情，发情次日屠宰、收集卵巢。应用IHC、WB检测卵巢组织Akt蛋白的定位和定量情况；RT-PCR检测Akt信号通路下游相关基因的表达量。结果表明：葡萄糖注射能够显著缓解限饲引起的延迟发情；0.5*M+Glu组P-Akt(473)蛋白的表达量显著高于（P < 0.05）0.5*M组与M组；0.5*M组GYS1、G6Pase的表达量均显著高于（P < 0.05）M组与0.5*M+Glu组；0.5*M+Glu组4EBPl的表达量显著高于（P < 0.05）M组与0.5*M组。初步推断：Akt信号通路参与调控黄体期营养水平对湖羊卵泡发育影响。30507
    毕业论文关键词：Akt信号通路；葡萄糖；卵泡发育
    Study on the Effects of Nutritional Level and Intravenous Injection of Glucose in Luteal Phase on Follicular Development of Hu Sheep on the Basis of Akt Signaling Pathway
    Abstract：To explore the molecular mechanism of the effect of different nutritional levels and intravenous injection of glucose in luteal phase on the follicular development of Hu sheep via Akt signaling pathway. Selected 24 purebred Hu sheep with similar body condition. At the 8th day of estrus cycle, all the sheep were randomly pided into three groups based on nutrient requirement recommended for maintenance(M), restricted feed group(0.5*M) and restricted feed + intravenous injection of glucose group(0.5*M+Glu). From 14th day of estrus cycle, estrus behavior was detected by testing the ewes with vasectomized rams. At the day after estrus, the sheep were slaughtered and ovaries collected. The localization and quantitative expression of Akt protein in ovarian tissue was detected by Immunohistochemistry and Western blotting. The expression of related genes of Akt signaling pathway downstream was detected by fluorescence quantitative PCR. The results of this study showed that: Intravenous injection of glucose could significantly ease the delayed estrus caused by feed restriction; The expression of P-Akt (473) protein of 0.5*M + Glu was significantly higher than(P <0.05)  that of 0.5*M and M groups; The expression of GYS1 and G6Pase of 0.5*M was significantly higher than (P <0.05) that of M and 0.5*M + Glu groups; The expression of 4EBPl of 0.5*M + Glu was significantly higher than(P <0.05) that of M and 0.5*M groups. The combined results indicated that Akt signaling pathway was involved in regulating the effect of the different nutritional levels and intravenous injection glucose in luteal phase on follicular development of Hu sheep.
    Key words: The Akt signaling pathway；glucose；follicular development；
    目  录
    摘要1
    关键词1
    Abstract 1
    Key words 1
    引言1
    1材料与方法2
    1.1试验动物 2
    1.2主要试剂和仪器 2
    1.3试验分组3
    1.4测定指标与方法 3
    1.4.1免疫组织化学分析3
    1.4.2蛋白印迹分析3
    1.4.3实时荧光定量分析4
    1.5数据统计分析4
    2结果与分析4
    2.1黄体期营养水平及静脉注射葡萄糖对湖羊发情周期的影响4
    2.2 Akt信号通路中Akt及P-Akt（473）蛋白的表达定位5
    2.3 Akt信号通路中Akt及P-Akt（473）蛋白表达量的检测5
    2.4黄体期营养水平及静脉注射葡萄糖对Akt信号通路下游基因表达量的影响6
    3讨论 6
    3.1黄体期营养水平及静脉注射葡萄糖对Akt信号通路中Akt蛋白磷酸化的影响6
    3.2黄体期营养水平及静脉注射葡萄糖对Akt信号通路下游相关基因表达量的影响7
    致谢7
    参考文献8
     基于Akt信号通路研究黄体期营养水平及静脉注射葡萄糖对湖羊卵泡发育的影响
    近年来，原产于太湖流域、具有多羔多产优良繁殖性状的湖羊越来越受到市场的青睐，北方传统牧区出现了湖羊的“引种热”，但湖羊在引入北方后，由于广大牧区冬季气温低，优质青饲料匮乏，母畜往往体质瘦弱，表现出乏情、排卵率低等不良状况，繁殖潜力受到严重制约。因此，针对湖羊营养性乏情开展繁殖调控技术的研发对提高湖羊繁殖力具有重要的意义。
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