GGTA1基因敲除巴马克隆猪与核供体巴马猪主要器官的比较解剖组织学研究

发布时间：2018-06-14 16:15

本文选题：GGTA1 + 基因敲除　；参考：《甘肃农业大学》2017年硕士论文

【摘要】：为了探讨GGTA1基因敲除巴马克隆猪与核供体巴马猪全身主要器官组织的解剖学与组织学发育变化异同,本试验选择GGTA1基因敲除巴马克隆猪2头(BM164和BM155)及核供体巴马猪1头(BMM2),利用大体解剖学方法对心脏、肝脏、脾脏、肺脏及肾脏进行了对比观察,利用组织学技术对大脑、小脑、心脏、皮肤、脾脏、淋巴结、甲状腺、肾上腺、脑垂体、食管、胃、肠、肝脏、胰腺、唾液腺、肺脏、肾脏以及输卵管和子宫等全身主要器官组织进行了比较研究,结果如下:1.解剖学比较结果核供体巴马猪心脏呈倒置圆锥形;肝脏为红褐色,可分为两面两缘四叶;脾脏为紫红色,镰刀形;肺脏呈海绵状粉红色,质软而轻,富有弹性;肾脏为平滑多乳头肾,左右对称,呈豆状。与核供体巴马猪相比,GGTA1基因敲除巴马克隆猪的心脏、肝脏、肺脏、肾脏及BM164号巴马猪脾脏的解剖学特征均无明显差异,但BM155号巴马猪脾脏有异常,其形态肿大且呈黑紫色。2.组织学比较结果与核供体巴马猪相比,GGTA1基因敲除巴马克隆猪全身主要器官的组织结构总体无显著差异,但也存在一些不同,具体如下:免疫系统中核供体巴马猪和BM164号巴马猪脾脏结构良好,发育正常,其内白髓和红髓分界明显,脾小体典型而丰富。但BM155号巴马猪脾脏脾小体组织结构不清晰,出血严重,嗜酸性粒细胞和中性粒细胞增多,网状纤维连接断续。消化系统中BM164号巴马猪空肠组织结构不完整,缺少肌层和浆膜;BM155号巴马猪肠管各段及胰腺出现了不同程度的结构变化,其十二指肠、空肠、盲肠、结肠的黏膜及黏膜下层内均可见较多的血细胞,结肠较发达,胰腺小叶间隔较大,间隔内可见大量脂肪组织,小叶被分割成数小块。BM164与BM155号巴马猪直肠均较BMM2号巴马猪发育良好。泌尿系统中核供体巴马猪肾脏结构良好。BM164和BM155号巴马猪肾脏组织结构总体完整,但在肾小球与肾小管中可见大量血细胞,肾小管上皮细胞出现轻微脱落坏死的现象。其它各系统器官组织包括大脑、小脑、心脏、皮肤、淋巴结、甲状腺、肾上腺、脑垂体食管、胃、肝脏、唾液腺、肺脏、输卵管及子宫等的组织结构均发育正常。上述结果提示利用现有的基因工程和动物克隆技术,可以得到主要器官组织发育正常的克隆巴马猪个体,并有望成为异种器官移植的良好平台与载体。但同时,本试验中克隆巴马猪个体的部分器官组织也出现了不同程度的结构变化,这可能与个体差异因素有关,其具体机制尚有待进一步研究和探讨。
[Abstract]:In order to study the anatomical and histological changes of the main organs and tissues of GGTA1 knockout cloned pig and nuclear donor pig, GGTA1 gene knockout Bama clone pigs (2 BM-164 and BM155) and nuclear donor BMM2P were used to observe the heart, liver, spleen, lung and kidney. The cerebellum and cerebellum were observed by histological technique. Heart, skin, spleen, lymph nodes, thyroid gland, adrenal gland, pituitary gland, esophagus, stomach, intestine, liver, pancreas, salivary gland, lung, kidney, oviduct and uterus were compared. The results were as follows: 1. Anatomical comparison showed that the heart of the nuclear donor was inverted conical; the liver was reddish brown and could be divided into two sides and four leaves; the spleen was purplish red and sickle shaped; the lung was spongy pink with soft light and elastic nature. Kidney is smooth polypapillary kidney, left and right symmetrical, beanlike. The anatomical characteristics of the heart, liver, lung, kidney and spleen of Bama pig with GGTA1 gene knockout were not significantly different from those of Bama pig, but the spleen of Bama pig BM155 was abnormal. There was no significant difference in the tissue structure of GGTA1 knockout major organs in the whole body compared with the nuclear donor Bama pig, but there were some differences in the tissue structure of the GGTA1 gene knockout pig. The main results are as follows: the spleen of Bama pig and BM164 pig in the immune system has good structure and normal development. The boundary between white pulp and red pulp is obvious and the splenic corpuscles are typical and abundant. However, the spleen splenic corpuscles of Bama pig BM155 were not clear, bleeding seriously, eosinophils and neutrophils increased, and reticular fibers were connected intermittently. In the digestive system, the jejunum of Bama pig BM164 was not complete, and there were some structural changes in the duodenum, jejunum and cecum of Bama pig without muscle layer, serosa BM155 and pancreas. More blood cells were found in the mucous membrane and submucous layer of the colon. The colon was more developed and the pancreatic interlobular septum was larger. A large amount of adipose tissue could be seen in the septum. The lobules were divided into several pieces. BM164 and BM155 Bama pig rectum developed better than BMM2 Bama pig. In the urinary system, the renal structure of the nuclear donor Bama pig. BM164 and BM155 were generally intact, but a large number of blood cells were found in the glomeruli and renal tubules, and the epithelial cells of the renal tubules showed slight exfoliation and necrosis. The tissues of other organs including brain cerebellum heart skin lymph node thyroid adrenal gland pituitary esophagus stomach liver salivary gland lung fallopian tube and uterus developed normally. These results suggest that the existing genetic engineering and animal cloning techniques can be used to clone Bama pig individuals with normal development of major organs and tissues, and it is expected to be a good platform and vector for xenotransplantation. But at the same time, some organs and tissues of Bama pig cloned in this experiment also showed different degree of structural changes, which may be related to individual differences, and its specific mechanism needs to be further studied and discussed.
【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S828

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