营养胁迫条件下牦牛皮下脂肪和背肌差异microRNAs的筛选与鉴定
发布时间：2018-08-18 08:49
【摘要】：牦牛(Bos gruniens)分布在青藏高原及其毗邻的高海拔地区,由于受高海拔条件及非地带性高原山地气候的制约,牦牛体内脂肪和肌肉呈现“波浪式”的消长方式。在牧草充足的暖季,牦牛体重迅速增长,体内肌肉及脂肪组织都得到快速生长与囤积;而在冷季,在长达7个月的枯草期后体重下降至暖季的70%左右,体内肌肉与脂肪组织均大量消耗。本研究采用高通量测序技术,研究冷季及暖季末期牦牛皮下脂肪及肌肉组织中差异表达的micro RNA及其功能,以探索micro RNA在营养胁迫下对牦牛脂肪及肌肉代谢平衡的调控作用。研究结果如下:1.构建冷季皮下脂、暖季皮下脂、冷季背最长肌以及暖季背最长肌四个小RNA文库,经Solexa测序得到的纯净读数均为24M。在四个文库中,长度为22 nt的micro RNA数量最多。2.在牦牛皮下脂肪micro RNA文库中,鉴定到450个micro RNA,其中70个micro RNA在两个季节皮下脂肪组织的表达量达到差异显著水平(P0.05)。在表达差异显著的micro RNA中,mi R-335、mi R-378b、mi R-200a、mi R-200b、mi R-200c及mi R-210对牦牛脂肪代谢起到调控作用。3.在牦牛背最长肌micro RNA文库中,鉴定到344个micro RNA。79个micro RNA在两个季节背最长肌组织中的表达量达到差异显著水平(P0.05),其中mi R-133b、mi R-378、mi R-378c、mi R-29a、mi R-29b和mi R-29c对牦牛肌肉代谢起到调控作用。4.皮下脂肪组织差异表达micro RNA预测到815个达到显著水平的靶基因,显著富集在脂肪细胞生成与代谢、脂肪酸的合成与分解等77个通路中(P0.05);背最长肌组织共预测到1315个达到显著水平的靶基因,靶基因在骨骼肌发育、蛋白质合成及氨基酸生成等99个通路中显著富集(P0.05)。综上所述,本研究通过Solexa高通量测序,筛选冷暖两季牦牛皮下脂肪及背最长肌组织micro RNAs并对其分子代谢通路进行研究,探索在冷季营养匮乏的胁迫下,micro RNAs对牦牛脂肪及肌肉代谢的重要调控作用,为诠释micro RNA在季节性环境变化下形成的特殊脂肪及肌肉代谢过程的调控作用提供了一定的科学依据和理论支撑。
[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.
【学位授予单位】：中国农业科学院
【学位级别】：博士
【学位授予年份】：2015
【分类号】：S823.85
[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.
【学位授予单位】：中国农业科学院
【学位级别】：博士
【学位授予年份】：2015
【分类号】：S823.85
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