本文選題：奶牛 + 乳腺��； 參考：《東北農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：牛是畜牧業(yè)中最重要的經(jīng)濟(jì)動物,牛奶的產(chǎn)量和質(zhì)量是衡量畜牧業(yè)水平和經(jīng)濟(jì)效益的一個重要標(biāo)準(zhǔn)。我國雖是世界奶牛養(yǎng)殖大國,但奶牛的平均單產(chǎn)世界的平均水平低。牛奶中的乳脂和乳蛋白的含量相對較低,對消費(fèi)者的消費(fèi)需求產(chǎn)生嚴(yán)重影響。乳汁是由乳腺腺泡的分泌細(xì)胞產(chǎn)生的。每個乳腺腺泡都由一層分泌上皮細(xì)胞構(gòu)成,這些高度特化的腺泡上皮細(xì)胞是奶牛乳腺泌乳的基本單位。腺泡上皮細(xì)胞一直持續(xù)增殖直達(dá)泌乳早期,乳腺分泌乳汁的能力由腺泡上皮細(xì)胞的數(shù)量決定。奶牛乳腺組織的發(fā)育及產(chǎn)生乳汁的過程當(dāng)中,一些重要基因的表達(dá)變化是調(diào)節(jié)腺泡上皮細(xì)胞分化、增殖以及乳汁合成、分泌、轉(zhuǎn)運(yùn)等生理過程的基礎(chǔ)。因此,篩選對奶牛乳腺組織的發(fā)育和產(chǎn)生乳汁有調(diào)節(jié)作用的基因以及鑒定這些基因的功能對提高奶牛乳腺的泌乳能力、促進(jìn)我國乳品業(yè)的發(fā)展有重要作用。脾源性酪氨酸激酶(spleen tyrosine kinase,SYK)是一種非受體型的蛋白酪氨酸激酶,在造血細(xì)胞、成纖維細(xì)胞、上皮細(xì)胞、肝細(xì)胞、血管內(nèi)皮細(xì)胞以及乳腺組織中都有所表達(dá)。SYK是一些細(xì)胞信號傳導(dǎo)途徑中關(guān)鍵的激酶,主要有調(diào)節(jié)細(xì)胞分化和增殖等作用。對于SYK基因的研究在乳腺癌案例中較多,但在正常的乳腺中研究較少。為探討SYK基因的表達(dá)與奶牛乳腺發(fā)育和泌乳功能之間的關(guān)系,本實(shí)驗(yàn)選取泌乳期高乳品質(zhì)、泌乳期低乳品質(zhì)和干乳期的中國荷斯坦奶牛,采用蛋白免疫印跡法和激光共聚焦顯微技術(shù)研究SYK基因在不同生理狀態(tài)和泌乳性狀的奶牛乳腺中的表達(dá)含量與表達(dá)部位變化;采用基因干擾和過表達(dá)技術(shù)研究SYK基因?qū)δ膛Ｈ橄偕掀ぜ?xì)胞增殖以及增殖相關(guān)信號分子表達(dá)的影響;采用雙熒光素酶報(bào)告基因系統(tǒng)研究SYK基因調(diào)節(jié)奶牛乳腺上皮細(xì)胞增殖的分子機(jī)制。Western blotting和激光共聚焦結(jié)果顯示干乳期奶牛乳腺組織中SYK的表達(dá)顯著高于泌乳期的奶牛乳腺組織。在干乳期的荷斯坦奶牛乳腺中,SYK主要在乳腺導(dǎo)管上皮細(xì)胞的胞質(zhì)中表達(dá)。而在泌乳期,SYK主要在奶牛乳腺腺泡上皮細(xì)胞的胞質(zhì)中表達(dá)�；蚋蓴_和過表達(dá)結(jié)果顯示SYK表達(dá)降低時,奶牛乳腺上皮細(xì)胞的增殖能力下降,β-酪蛋白的表達(dá)量降低,同時與細(xì)胞增殖相關(guān)信號分子AKT1和MAPK等蛋白表達(dá)量也相對降低;而SYK過表達(dá)時,細(xì)胞增殖增加、β-酪蛋白、AKT1和MAPK的蛋白表達(dá)量上調(diào)。雙熒光素酶報(bào)告基因系統(tǒng)研究結(jié)果顯示奶牛乳腺上皮細(xì)胞中SYK可以激活A(yù)KT1啟動子的活性,其作用位點(diǎn)位于AKT1啟動子區(qū)-439~+83bp的序列范圍。這些結(jié)果表明SYK是奶牛乳腺上皮細(xì)胞增殖與分化的調(diào)節(jié)因子,SYK基因通過調(diào)節(jié)AKT1的轉(zhuǎn)錄活性促進(jìn)奶牛乳腺上皮細(xì)胞增殖,為干乳期乳腺組織重建做準(zhǔn)備。這項(xiàng)研究結(jié)果可以豐富泌乳生物學(xué)理論,為提高牛奶產(chǎn)量和質(zhì)量提供一定的理論依據(jù),對促進(jìn)我國乳產(chǎn)業(yè)的進(jìn)一步發(fā)展具有重要的理論意義和生產(chǎn)實(shí)際意義。
[Abstract]:Cattle are the most important economic animals in animal husbandry. The yield and quality of milk are an important standard to measure the level of animal husbandry and economic benefits. Although China is a big country in the world, the average level of the average yield of dairy cows is low. The content of milk fat and milk protein is relatively low, and the consumption demand of consumers is produced. "Milk is produced by the secretory cells of the mammary glands. Each mammary gland is made up of a layer of epithelial cells. These highly specialized acinar epithelial cells are the basic units of milk secretion in the cow's mammary glands. The acinar epithelial cells continue to proliferate directly to the early lactation, and the ability of the mammary glands to secrete milk from the acinar epithelial cells." Quantitative determination. During the development of mammary gland tissue and the process of milk production, the expression of some important genes is the basis for regulating the differentiation and proliferation of acinar epithelial cells, and the basis of physiological processes such as milk synthesis, secretion, transport and other physiological processes. Therefore, the screening of genes that regulate the development of mammary gland tissue and the regulation of milk production and identification of milk cow's mammary gland tissue and identification of this The function of some genes plays an important role in improving the lactating ability of dairy cows and promoting the development of dairy industry in China. Spleen tyrosine kinase (SYK) is a kind of non receptor protein tyrosine kinase, which is found in hematopoietic cells, fibroblasts, epithelial cells, hepatocytes, vascular endothelial cells, and mammary tissues. The expression of.SYK is the key kinase in some cell signaling pathways, which mainly regulate cell differentiation and proliferation. The study of SYK gene is more in breast cancer cases, but less in normal mammary gland. In order to explore the relationship between the expression of SYK gene and the mammary gland development and lactation function of dairy cows, this experiment selected secretion. Chinese Holstein cows with high milk quality, low lactation quality and dry milk stage were studied by protein immunoblotting and laser confocal microscopy to study the expression and expression of SYK gene in dairy cows with different physiological status and lactation traits, and SYK gene was used to study the milk of milk in the milk of dairy cows with different physiological States and lactation traits. The effect of the proliferation of bovine mammary epithelial cells and the expression of proliferation related signal molecules; the molecular mechanism of SYK gene regulating the proliferation of mammary epithelial cells by double luciferase reporter gene system.Western blotting and laser confocal results showed that the expression of SYK in the mammary tissues of dairy cows with dry milk period was significantly higher than that of lactating cows. SYK is mainly expressed in the cytoplasm of the mammary ductal epithelial cells during the dry milk period of Holstein cows. While in lactation, SYK is mainly expressed in the cytoplasm of the mammary gland epithelial cells of the dairy cows. The results of gene interference and overexpression show that the proliferation of milk gland epithelial cells in dairy cows decreased when the expression of SYK decreased, and the beta casein was reduced. At the same time, the expression of AKT1 and MAPK related to cell proliferation related signal molecules also decreased; while SYK overexpression, the proliferation of cells increased, the protein expression of beta casein, AKT1 and MAPK was up-regulated. The results of the double luciferase reporter gene system study showed that SYK could activate the AKT1 promoter in milk bovine mammary epithelial cells. The activity, its site of action, is located in the range of -439~+83bp in the promoter region of AKT1. These results suggest that SYK is a regulator of the proliferation and differentiation of mammary epithelial cells in dairy cows. The SYK gene promotes the proliferation of mammary epithelial cells in dairy cows by regulating the transcriptional activity of AKT1. The results of this study can enrich the secretion of mammary glands in the dry milk. The theory of milk biology provides a certain theoretical basis for improving the production and quality of milk. It is of great theoretical significance and practical significance to promote the further development of dairy industry in China.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S823

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

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