【摘要】：牦牛(Bos gruniens)分布在青藏高原及其毗鄰的高海拔地區(qū),由于受高海拔條件及非地帶性高原山地氣候的制約,牦牛體內(nèi)脂肪和肌肉呈現(xiàn)“波浪式”的消長方式。在牧草充足的暖季,牦牛體重迅速增長,體內(nèi)肌肉及脂肪組織都得到快速生長與囤積;而在冷季,在長達7個月的枯草期后體重下降至暖季的70%左右,體內(nèi)肌肉與脂肪組織均大量消耗。本研究采用高通量測序技術(shù),研究冷季及暖季末期牦牛皮下脂肪及肌肉組織中差異表達的micro RNA及其功能,以探索micro RNA在營養(yǎng)脅迫下對牦牛脂肪及肌肉代謝平衡的調(diào)控作用。研究結(jié)果如下:1.構(gòu)建冷季皮下脂、暖季皮下脂、冷季背最長肌以及暖季背最長肌四個小RNA文庫,經(jīng)Solexa測序得到的純凈讀數(shù)均為24M。在四個文庫中,長度為22 nt的micro RNA數(shù)量最多。2.在牦牛皮下脂肪micro RNA文庫中,鑒定到450個micro RNA,其中70個micro RNA在兩個季節(jié)皮下脂肪組織的表達量達到差異顯著水平(P0.05)。在表達差異顯著的micro RNA中,mi R-335、mi R-378b、mi R-200a、mi R-200b、mi R-200c及mi R-210對牦牛脂肪代謝起到調(diào)控作用。3.在牦牛背最長肌micro RNA文庫中,鑒定到344個micro RNA。79個micro RNA在兩個季節(jié)背最長肌組織中的表達量達到差異顯著水平(P0.05),其中mi R-133b、mi R-378、mi R-378c、mi R-29a、mi R-29b和mi R-29c對牦牛肌肉代謝起到調(diào)控作用。4.皮下脂肪組織差異表達micro RNA預(yù)測到815個達到顯著水平的靶基因,顯著富集在脂肪細胞生成與代謝、脂肪酸的合成與分解等77個通路中(P0.05);背最長肌組織共預(yù)測到1315個達到顯著水平的靶基因,靶基因在骨骼肌發(fā)育、蛋白質(zhì)合成及氨基酸生成等99個通路中顯著富集(P0.05)。綜上所述,本研究通過Solexa高通量測序,篩選冷暖兩季牦牛皮下脂肪及背最長肌組織micro RNAs并對其分子代謝通路進行研究,探索在冷季營養(yǎng)匱乏的脅迫下,micro RNAs對牦牛脂肪及肌肉代謝的重要調(diào)控作用,為詮釋micro RNA在季節(jié)性環(huán)境變化下形成的特殊脂肪及肌肉代謝過程的調(diào)控作用提供了一定的科學(xué)依據(jù)和理論支撐。
[Abstract]:Yak (Bos gruniens) is distributed in the Qinghai-Tibet Plateau and its adjacent high altitude area. Because of the restriction of high altitude and non-zonal plateau mountain climate, the fat and muscle of yak show a "wave-like" growth and decline pattern. During the warm season when the forage is plentiful, yaks gain weight rapidly, their muscles and fat tissue grow and hoard rapidly, while in the cold season, after seven months of dry grass, the weight drops to about 70 percent of the warm season. Muscle and adipose tissue in the body are consumed in large quantities. The purpose of this study was to investigate the differential expression of micro RNA and its function in subcutaneous fat and muscle tissue of yaks in cold season and warm season by using high throughput sequencing technique to explore the regulation of micro RNA on metabolism balance of fat and muscle in yak under nutritional stress. The results are as follows: 1. Four small RNA libraries were constructed: subcutaneous lipid in cold season, subcutaneous lipid in warm season, longissimus dorsi muscle in cold season and longissimus dorsi muscle in warm season. The pure reading obtained by Solexa sequencing was 24M. Of the four libraries, 22 NT in length of micro RNA is the largest. 2. 2. 450 micro RNAs were identified in yak subcutaneous fat micro RNA library and 70 micro RNA expression levels reached significant level in subcutaneous adipose tissue of two seasons (P0.05). In the micro RNA with significant difference in expression, mi-mi R-335n mi R-378bmnmi R-200aanmi R-200bnmmiR-200c and miR-210 play a role in regulating the fat metabolism of yak. In the micro RNA library of the longissimus dorsi muscle of yak, the expression of 344 micro RNA.79 micro RNA in the longissimus dorsi muscle was significantly different between the two seasons (P0.05). Among them, the expression of the micro RNA in the longissimus dorsi muscle of yak was significantly higher than that in the other two seasons (P0.05). Among them, the expression of the micro RNA in the longissimus dorsi muscle of yak was significantly different (P0.05). Differential expression of micro RNA in subcutaneous adipose tissue predicted 815 target genes, which were significantly enriched in adipogenesis and metabolism. Fatty acid synthesis and decomposition were found in 77 pathways (P0.05), and 1315 target genes were predicted to reach significant level in longissimus dorsi (P0.05). Target genes were significantly enriched in 99 pathways such as skeletal muscle development, protein synthesis and amino acid production (P0.05). In conclusion, Solexa high-throughput sequencing was used to screen micro RNAs from subcutaneous fat and longissimus dorsi muscle of cold and warm yaks and to study its molecular metabolic pathway. To explore the important regulation effect of micro RNAs on fat and muscle metabolism in yak under the stress of nutrition deficiency in cold season. It provides a scientific basis and theoretical support for explaining the regulation of the metabolism process of special fat and muscle formed by micro RNA in seasonal environment.
【學(xué)位授予單位】：中國農(nóng)業(yè)科學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：S823.85

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