本文選題：牦牛 + 器官組織　； 參考：《甘肅農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：本研究旨在探究生長激素受體(growth hormone receptor,GHR)在牦牛不同器官組織中分布與表達(dá)情況。采集成年牦牛的不同器官組織(心臟、肝臟、脾臟、肺臟、腎臟和睪丸),從各組織中提取總RNA,并通過反轉(zhuǎn)錄合成第一鏈cDNA,然后根據(jù)GeneBank中家牛GHR基因序列和牦牛β-actin的基因序列設(shè)計(jì)特異性引物。采用實(shí)時(shí)熒光定量PCR測定牦牛GHR基因在不同組織中的相對(duì)表達(dá)量。同時(shí),用4%的多聚甲醛溶液固定組織樣品并制成石蠟切片,采用免疫組織化學(xué)法檢測GHR蛋白在不同組織中的分布及定位,用Image-Pro-Plus 6.0分析GHR陽性反應(yīng)物的吸光度,并測定其積分光密度值(IOD),然后在SPSS 19.0中用單因素方差法對(duì)各積分光密度值進(jìn)行差異顯著性分析。研究結(jié)果表明:(1)GHR基因在牦牛不同器官組織中的表達(dá)存在差異性,肝臟中的相對(duì)表達(dá)量最高,分別是心臟、腎臟、睪丸、肺臟和脾臟中的3.73、4.98、5.03、8.58和10.21倍。(2)GHR蛋白在肝臟、心臟、腎臟、睪丸、肺臟和脾臟中均有分布,且陽性反應(yīng)強(qiáng)度不等。在睪丸的曲精小管、細(xì)支氣管上皮細(xì)胞、脾小結(jié)生發(fā)中心、卵母細(xì)胞和顆粒細(xì)胞的細(xì)胞質(zhì)中可觀察到較明顯的陽性反應(yīng),細(xì)胞核反應(yīng)極少;而在肝細(xì)胞、心肌細(xì)胞以及腎小管上皮細(xì)胞中,胞質(zhì)和胞核均有GHR的強(qiáng)烈表達(dá)。(3)肝臟中GHR蛋白的表達(dá)最高,心臟、腎臟、睪丸和脾臟次之,肺臟中最少。由上述可以看出,肝臟中GHR的基因和蛋白表達(dá)量均要高于心臟、脾臟、肺臟、腎臟和睪丸,推測與肝臟參與調(diào)控動(dòng)物機(jī)體生長代謝的重要作用有關(guān)。本實(shí)驗(yàn)首次在基因水平和蛋白水平表明了GHR在牦牛不同器官組織中的表達(dá)具有差異性,為牦牛的改良和育種提供了一定的理論資料。
[Abstract]:The purpose of this study was to investigate the distribution and expression of growth hormone receptor (GHR) in different organs of yak. The different organs and tissues (heart, liver, spleen, lung) of adult yaks were collected. In kidney and testis, total RNAs were extracted from various tissues, and the first strand of cDNAs was synthesized by reverse transcription. Specific primers were designed according to the sequence of GHR gene of domestic cattle and the gene sequence of 尾 -actin of yak in GeneBank. The relative expression of yak GHR gene in different tissues was measured by real-time fluorescence quantitative PCR. At the same time, the tissue samples were fixed with 4% paraformaldehyde solution and made into paraffin sections. The distribution and localization of GHR protein in different tissues were detected by immunohistochemical method. The absorbance of GHR positive reactants was analyzed by Image-Pro-Plus 6.0. The integral optical density (IOD) was measured, and the difference of the integral optical density was analyzed by single factor variance method in SPSS 19.0. The results showed that there were differences in the expression of GHR gene in different organs of yak, and the highest relative expression of GHR in liver was found in heart, kidney, testis, lung and spleen. The expression of GHR protein in liver, heart and kidney was 8.58 and 10.21 times higher than that in liver, heart, testis, lung and spleen. Testis, lungs and spleen were all distributed, and the intensity of positive reaction varied. In testicular seminiferous tubules, bronchiolar epithelial cells, germinal centers of splenic nodules, oocytes and granulosa cells, the cytoplasm of oocytes and granulosa cells showed more obvious positive reactions. In cardiomyocytes and tubular epithelial cells, GHR was strongly expressed in cytoplasm and nucleus. The expression of GHR protein was the highest in liver, followed by heart, kidney, testis and spleen, and the least in lung. It can be seen that the expression of GHR gene and protein in liver is higher than that in heart, spleen, lung, kidney and testis, which may be related to the important role of liver in regulating the growth and metabolism of animal body. This experiment for the first time showed that the expression of GHR in different organs and tissues of yaks was different at the gene and protein levels, which provided some theoretical data for the improvement and breeding of yaks.
【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S823.85

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相關(guān)期刊論文 前1條

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本文編號(hào)：1922089