本文選題：鴨 + T3�。� 參考：《中國農(nóng)業(yè)科學(xué)院》2015年碩士論文

【摘要】：胚胎期骨骼肌的發(fā)生決定鴨產(chǎn)肉量,比較發(fā)現(xiàn)生長速度差異較大兩品種鴨胚胎期生長速度不同,且生長速度較快品種鴨蛋清內(nèi)三碘甲腺原氨酸(T3)含量顯著高于后者。假設(shè)蛋清內(nèi)T3含量與鴨生長速度呈正相關(guān),增加蛋清內(nèi)T3含量會促進(jìn)鴨生長。為驗(yàn)證假說,設(shè)計(jì)蛋清內(nèi)注射T3試驗(yàn),監(jiān)測鴨出雛前后生長、血清中甲狀腺激素(thyroid hormones,THs)含量及THs穩(wěn)態(tài)的主要調(diào)控因子--肝臟中脫碘酶(deiodinases)的表達(dá)情況;設(shè)計(jì)外源性T3干擾鴨胚原代肝細(xì)胞生長試驗(yàn),檢測細(xì)胞中脫碘酶的表達(dá)。探討外源性T3對鴨出雛前后生長及肝臟中脫碘酶表達(dá)的影響。1蛋清內(nèi)注射T3對鴨出雛前后生長及血清中THs含量的影響金定鴨種蛋200枚,隨機(jī)分為兩組:對照組蛋清內(nèi)(Control group,Con)注射100μL不含T3的生理鹽水,試驗(yàn)組蛋清內(nèi)(Treatment group,Tre)注射100μL含250 ng T3的生理鹽水,兩組種蛋在同一條件下同期孵化。監(jiān)測27胚齡和7日齡鴨體重和胸肌重的變化,并采集鴨的血液;放射性免疫分析法檢測血清中T3和T4的含量;分析鴨出雛前后體重、胸肌重與血清T3、T4含量之間的相關(guān)性。結(jié)果表明,試驗(yàn)組鴨體重和胸肌重在27胚齡和7日齡均顯著高于對照組(P=0.035,0.044;0.027,0.037),試驗(yàn)組血清中T3和T4的含量在7日齡均顯著低于對照組(P=0.040,0.040);試驗(yàn)組鴨體重與T3含量呈極顯著負(fù)線性相關(guān)(r=-0.821,P=0.000)。2蛋清內(nèi)注射T3對鴨出雛前后肝臟中脫碘酶表達(dá)的影響試驗(yàn)設(shè)計(jì)與上一章相同,采集鴨27胚齡和7日齡時的肝臟。實(shí)時熒光定量PCR(RT-PCR)檢測肝臟中D1和D3的mRNA表達(dá),ELISA法檢測肝臟中D1和D3的蛋白含量及酶活性。分析D1和D3的表達(dá)與血清T3、T4含量之間的相關(guān)性。結(jié)果表明,試驗(yàn)組肝臟D3的mRNA表達(dá)以及D1、D3的蛋白含量和酶活性在7日齡均顯著低于對照組(P=0.049;0.017,0.042;0.013,0.039)。試驗(yàn)組血清中T4含量與D3的mRNA表達(dá)呈極顯著正線性相關(guān)(r=0.913,P=0.001),T3含量與D1和D3蛋白含量及酶活性均呈顯著正線性相關(guān)(r=0.790,P=0.011;r=0.746,P=0.021;r=0.757,P=0.018;r=0.744,P=0.022)。3外源性T3對鴨胚原代肝細(xì)胞中脫碘酶表達(dá)的影響分離并培養(yǎng)鴨15胚齡原代肝細(xì)胞,24 h后隨機(jī)分為三組即對照組(Control group,Con)、低濃度T3干擾組(Low T3 group,LT3)和高濃度T3干擾組(High T3 group,HT3)。Con組細(xì)胞液中導(dǎo)入2μL不含T3的生理鹽水;LT3和HT3組細(xì)胞液中分別導(dǎo)入終濃度為5 ng/mL和20 ng/mL的T3,均以2μL生理鹽水為載體。干擾36 h后細(xì)胞換液并繼續(xù)培養(yǎng)12 h。在干擾后0 h和12 h時收集細(xì)胞并用Cell Counting Kit(CCK-8)法檢測細(xì)胞活性,RT-PCR檢測細(xì)胞中D1和D3的mRNA表達(dá),ELISA檢測細(xì)胞D1和D3的蛋白含量及酶活性。結(jié)果顯示,LT3組細(xì)胞活性及D1的mRNA表達(dá)在干擾后的0 h和12 h均顯著高于對照組(P=0.026,0.006;0.000,0.000)。全文結(jié)論:蛋清內(nèi)T3含量的差異與鴨的早期生長有一定的相關(guān)性,外源性T3(250 ng)促進(jìn)了鴨出雛前后體重和胸肌重的增長,降低了鴨出雛后血清中T3和T4的含量;鴨出雛前后生長模式發(fā)生改變,伴隨著出雛前D3 mRNA表達(dá)的下調(diào)及出雛后D1和D3的蛋白含量及酶活性的降低;外源性T3干擾鴨胚原代肝細(xì)胞生長試驗(yàn)驗(yàn)證了D1對T3的直接調(diào)控作用。
[Abstract]:The amount of meat duck embryo determining skeletal muscle, that growth rate differences between the two varieties of duck embryo growth rate and faster growth of different varieties of duck's egg white three triiodothyronine (T3) was significantly higher than that of the latter. The content of T3 in duck egg white that growth rate was positively correlated with increased T3 content in egg white will promote growth. In order to verify the hypothesis of duck egg white, design of T3 injected into the test, monitoring duck hatching and growth of thyroid hormone in serum (thyroid, hormones, THs) deiodinase regulating factor of liver and the content of THs in steady state (deiodinases) the expression of exogenous T3 interference; design of duck embryo primary liver cell growth test, expression of deiodinases were detected. To investigate the effect of exogenous T3 on deiodinase expression before and after hatching and growth of duck liver.1 in egg white duck hatching before and after the injection of T3 on growth and serum THs content The influence of Jinding duck eggs 200, were randomly divided into two groups: control group (Control, group, Con in egg white) saline injection 100 L without T3, the test group (Treatment group, Tre in egg white) saline injection 100 L containing 250 ng T3, two groups of eggs in the same a period of incubation. 27 monitoring conditions below and 7 day old duck embryo weight and breast muscle weight changes, and duck blood collection; analysis of the content of T3 and T4 in serum were determined by radioactive immune analysis; duck hatching and weight, breast muscle weight and serum T3, the correlation between T4 content. The results show that the weight the test group 27 and chest on duck embryo age and 7 days of age were significantly higher than those in control group (P=0.035,0.044; 0.027,0.037), the content of T3 and T4 in serum of the experimental group at 7 days of age were significantly lower than the control group (P=0.040,0.040); experimental group duck weight and T3 content showed significant negative correlation (r= -0.821, P=0.000.2 injection of T3 in egg white) Effect on duck deiodinase expression in the liver before and after the young experimental design and the same chapter, collecting duck 27 and 7 day old embryo liver. Real time fluorescence quantitative PCR (RT-PCR) to detect the expression of D1 and D3 in the liver of mRNA, protein content and enzyme activity of D1 and D3 detection in liver by ELISA method the expression and analysis of serum T3. D1 and D3, the correlation between the content of T4. The results show that the experimental group of liver D3 mRNA expression and D1, protein content and enzyme activity of D3 at 7 days of age were significantly lower than the control group (P=0.049; 0.017,0.042; 0.013,0.039). The content of T4 in serum and D3 in test group the expression of mRNA had a significantly positive linear correlation (r=0.913, P=0.001), showed a significant positive linear correlation with T3 content of D1 and D3 protein content and enzyme activity (r=0.790, P=0.011; r=0.746, P=0.021; r=0.757, P=0.018; r=0.744, P=0.022) separation effect of deiodinase expression in primary liver cells.3 exogenous T3 the duck embryo 騫跺煿鍏婚腑15鑳氶緞鍘熶唬鑲濈粏鑳，

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