水牛泌乳期与非泌乳期乳腺组织miRNAs表达谱鉴定及差异表达分析

doi:10.11843/j.issn.0366-6964.2017.09.008

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (9): 1635-1647.doi: 10.11843/j.issn.0366-6964.2017.09.008

水牛泌乳期与非泌乳期乳腺组织miRNAs表达谱鉴定及差异表达分析

蔡小艳^1,2, 鲍正攀¹, 古景开¹, 邓凯¹, 张晓溪¹, 任艳萍¹, 沈朋雷¹, 石德顺^1*, 刘庆友^1*

1. 广西大学亚热带农业生物资源保护与利用国家重点实验室, 南宁 530004;
2. 广西壮族自治区畜牧研究所, 南宁 530001

收稿日期:2017-01-09 出版日期:2017-09-23 发布日期:2017-09-23
通讯作者: 石德顺,博士,研究员,E-mail:ardsshi@gxu.edu.cn;刘庆友,博士,研究员,主要从事动物生殖生理研究,E-mail:qyliu2002@126.com
作者简介:蔡小艳(1982-),女,山西兴县人,博士生,主要从事牧草及草食动物遗传育种与推广应用,E-mail:caixiaoyan282@163.com
基金资助:
国家863重点项目课题（2013AA102504）：广西科技厅项目（桂科合14125008-2-13；桂科AD17129043；桂科AA16380026）

Identify and Analyze the Expression Profile and Mechanism of Buffalo Mammary Gland miRNAs in the Lactation and Non-lactation Periods

CAI Xiao-yan^1,2, BAO Zheng-pan¹, GU Jing-kai¹, DENG Kai¹, ZHANG Xiao-xi¹, REN Yan-ping¹, SHEN Peng-lei¹, SHI De-shun^1*, LIU Qing-you^1*

1. State Key Laboratory of Subtropical Bioresource Conservation and Utilization, Guangxi University, Nanning 530004, China;
2. Guangxi Institute of Animal Science, Nanning 530001, China

Received:2017-01-09 Online:2017-09-23 Published:2017-09-23

摘要/Abstract

摘要：

旨在鉴定分析水牛乳腺泌乳期和非泌乳期miRNA的表达谱和差异作用机制。采集水牛泌乳期和非泌乳期乳腺组织，利用Solexa测序技术构建2个时期miRNAs表达谱，鉴定前体miRNAs、成熟miRNAs和新miRNAs，分析miRNAs的第一偏好性核苷酸和染色体分布，发掘水牛泌乳期和非泌乳期高表达及差异表达miRNAs。结果显示，本研究构建了水牛泌乳期和非泌乳期乳腺组织2个miRNA表达谱，分别获得12 768 110和12 569 467条18~31 nt的高质量序列。测序确认了归属259个miRNAs家族的359个成熟miRNAs和363个pre-miRNAs及5个水牛特有的miRNAs。U是19和25 nt miRNAs的5'端最普遍的核苷酸。bbu-let-7b、bbu-let-7a、miR-26a和miR-21等miRNAs在2个时期均呈现高表达；bbu-miR-148a、143、200c、200a和bbu-let-7f等miRNAs在非泌乳期特异性高表达。bbu-miR-125b、29a和bbu-let-7c等miRNAs在泌乳期特异性高表达。bbu-miR-148a、143、200a、141和30a-5p等miRNAs在泌乳期的表达量下降为非泌乳期的1/2以下。而bbu-miR-26a、29a、125b、99a和bbu-let-7c等miRNAs在泌乳期表达量大于或等于2倍非泌乳期丰度。本研究构建了水牛泌乳期以及非泌乳期乳腺组织2个miRNAs表达谱，测序确认了259个水牛miRNAs家族的359个成熟miRNAs和363个前体miRNAs，5个水牛特有miRNAs和10个高差异表达miRNAs，为进一步阐明奶水牛泌乳关键miRNAs作用机制奠定了基础。

Abstract:

This study aimed to identify and analyze the expression patterns and mechanism of buffalo mammary gland miRNAs in the lactation and non-lactation periods. Buffalo mammary gland tissues in lactation and non-lactation periods were collected, their miRNAs expression profiles were analyzed by Solexa sequencing technology, pre-miRNAs, mature miRNAs and novel miRNAs were identified, respectively, the first preferred nucleotide and chromosome distribution were analyzed. The highly and differentially expressed miRNAs in the lactation and non-lactation periods were detected. The results showed that 2 miRNAs expression profiles from buffalo lactation and non-lactation mammary glands were constructed. 12 768 110 and 12 569 467 high-quality reads between 18 and 31 nt were obtained seperately. Three hundred fifty-nine mature miRNAs and 363 pre-miRNAs belonged to 259 miRNAs families, and 5 buffalo-specific miRNAs were confirmed by sequencing. U was the most common nucleotide at the 5'end of 19 and 25 nt miRNAs. The bbu-let-7b, bbu-let-7a, miR-26a and miR-21 showed high expression in both periods. bbu-miR-148a, 143, 200c, 200a and bbu-let-7f were specificially highly expressed in non-lactation period, the bbu-miR-125b, 29a and bbu-let-7c were specifically highly expressed in lactation period. The bbu-miR-148a, 143, 200a, 141 and 30a-5p decreased the expression in the lactation period to less than 1/2 of the non-lactation period, bbu-miR-26a, 29a, 125b, 99a and bbu-let-7c were highly expressed in the lactation period more than or equal to 2 times of the lactation period. The result indicate that 2 miRNAs profiles from buffalo lactation and non-lactation mammary gland tissues were constructed. 359 mature miRNAs and 363 pre-miRNAs belonging to 259 buffalo miRNAs families were identified. 5 buffalo genome-specific miRNAs and 10 highly differentially expressed miRNAs were obtained, which lay the foundation for further elucidating the mechanism of key miRNAs in lactating buffalo.

中图分类号:

S823.83

蔡小艳, 鲍正攀, 古景开, 邓凯, 张晓溪, 任艳萍, 沈朋雷, 石德顺, 刘庆友. 水牛泌乳期与非泌乳期乳腺组织miRNAs表达谱鉴定及差异表达分析[J]. 畜牧兽医学报, 2017, 48(9): 1635-1647.

CAI Xiao-yan, BAO Zheng-pan, GU Jing-kai, DENG Kai, ZHANG Xiao-xi, REN Yan-ping, SHEN Peng-lei, SHI De-shun, LIU Qing-you. Identify and Analyze the Expression Profile and Mechanism of Buffalo Mammary Gland miRNAs in the Lactation and Non-lactation Periods[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(9): 1635-1647.

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水牛泌乳期与非泌乳期乳腺组织miRNAs表达谱鉴定及差异表达分析

Identify and Analyze the Expression Profile and Mechanism of Buffalo Mammary Gland miRNAs in the Lactation and Non-lactation Periods

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