Comparative transcript profiling of gene expression of fresh and frozen-thawed bull sperm
Comparative transcript profiling of gene expression of fresh and frozen-thawed bull sperm
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DOI：
10.1016/j.theriogenology.2014.10.015
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发表时间：
2015-03-01
期刊：
影响因子：
2.8
通讯作者：
Wang, Dong
中科院分区：
文献类型：
--
作者：
Chen, Xiaoli;Wang, Yonggui;Wang, Dong
尽管冷冻精液在商业上广泛应用于养牛行业，但由此产生的怀孕率低于使用新鲜精液产生的怀孕率。冷冻损伤是精液冷冻保存的主要问题。它会导致精子转录物的改变，从而影响精子的功能和活力。我们利用抑制减法杂交技术建立互补DNA减法文库，结合微阵列技术和序列同源性分析对文库中差异表达基因进行筛选分析，并将9头公牛的新鲜精子与冻融精子进行比较。总体而言，通过微阵列数据和Microarrays软件的显著性分析（vertical bar score (d)竖线>= 2,fold change > 1，假发现率< 0.05）鉴定出19个阳性差异表达的unique。15个差异表达的单基因序列同源性高（e值为1E(-03)），被认为是新基因。其中5个基因——rpl31、PRKCE、PAPSS2、PLP1和r1g7的表达通过实时定量逆转录聚合酶链反应验证。RPL31基因表达差异有统计学意义（P < 0.05）。我们的初步结果提供了荷斯坦公牛的新鲜和冷冻解冻精子之间差异表达转录物的概述。(C) 2015爱思唯尔公司版权所有。
Although frozen semen is widely used commercially in the cattle breeding industry, the resultant pregnancy rate is lower than that produced using fresh semen. Cryodamage is a major problem in semen cryopreservation; it causes changes to sperm transcripts that may influence sperm function and motility. We used suppression subtractive hybridization technology to establish a complementary DNA subtractive library, and combined microarray technology and sequence homology analysis to screen and analyze differentially expressed genes in the library, comparing fresh sperm with the frozen-thawed sperm of nine bulls. Overall, 19 positive differentially expressed unigenes were identified using microarray data and Significance Analysis of Microarrays software (vertical bar score (d)vertical bar >= 2, fold change > 1, and false discovery rate < 0.05). Of 15 differentially expressed unigenes exhibited high sequence homology (E-value 1E(-03)) and were considered novel genes. The expression of five of these genes-RPL31, PRKCE, PAPSS2, PLP1, and R1G7-was verified by quantitative real-time reverse transcription-polymerase chain reaction. There was a significant differential expression of the RPL31 gene (P < 0.05). Our preliminary results provide an overview of differentially expressed transcripts between fresh and frozen-thawed sperm of Holstein bulls. (C) 2015 Elsevier Inc. All rights reserved.