生长激素受体（GHR）在牦牛不同器官组织中的分布与表达

发布时间：2018-05-22 12:30

本文选题：牦牛 + 器官组织　；参考：《甘肃农业大学》2015年硕士论文

【摘要】：本研究旨在探究生长激素受体(growth hormone receptor,GHR)在牦牛不同器官组织中分布与表达情况。采集成年牦牛的不同器官组织(心脏、肝脏、脾脏、肺脏、肾脏和睾丸),从各组织中提取总RNA,并通过反转录合成第一链cDNA,然后根据GeneBank中家牛GHR基因序列和牦牛β-actin的基因序列设计特异性引物。采用实时荧光定量PCR测定牦牛GHR基因在不同组织中的相对表达量。同时,用4%的多聚甲醛溶液固定组织样品并制成石蜡切片,采用免疫组织化学法检测GHR蛋白在不同组织中的分布及定位,用Image-Pro-Plus 6.0分析GHR阳性反应物的吸光度,并测定其积分光密度值(IOD),然后在SPSS 19.0中用单因素方差法对各积分光密度值进行差异显著性分析。研究结果表明:(1)GHR基因在牦牛不同器官组织中的表达存在差异性,肝脏中的相对表达量最高,分别是心脏、肾脏、睾丸、肺脏和脾脏中的3.73、4.98、5.03、8.58和10.21倍。(2)GHR蛋白在肝脏、心脏、肾脏、睾丸、肺脏和脾脏中均有分布,且阳性反应强度不等。在睾丸的曲精小管、细支气管上皮细胞、脾小结生发中心、卵母细胞和颗粒细胞的细胞质中可观察到较明显的阳性反应,细胞核反应极少;而在肝细胞、心肌细胞以及肾小管上皮细胞中,胞质和胞核均有GHR的强烈表达。(3)肝脏中GHR蛋白的表达最高,心脏、肾脏、睾丸和脾脏次之,肺脏中最少。由上述可以看出,肝脏中GHR的基因和蛋白表达量均要高于心脏、脾脏、肺脏、肾脏和睾丸,推测与肝脏参与调控动物机体生长代谢的重要作用有关。本实验首次在基因水平和蛋白水平表明了GHR在牦牛不同器官组织中的表达具有差异性,为牦牛的改良和育种提供了一定的理论资料。
[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.
【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S823.85

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