Comparative transcript profiling of gene expression of fresh and frozen-thawed bull sperm

被引:60
作者
Chen, Xiaoli [1 ]
Wang, Yonggui [2 ]
Zhu, Huabin [1 ]
Hao, Haisheng [1 ]
Zhao, Xueming [1 ]
Qin, Tong [1 ]
Wang, Dong [1 ]
机构
[1] Chinese Acad Agr Sci, Inst Anim Sci, Minist Agr China, Key Lab Farm Anim Genet Resources & Utilizat, Beijing 100193, Peoples R China
[2] Jilin Agr Univ, Changchun 130118, Peoples R China
关键词
Bull; Fresh and frozen-thawed sperm; Differentially expressed genes; SSH; cDNA microarray; SUPPRESSION-SUBTRACTIVE HYBRIDIZATION; PROTEIN-KINASE-C; BOAR SPERMATOZOA; NONCODING RNAS; MESSENGER-RNA; SEMEN; CRYOPRESERVATION; SPERMATOGENESIS; IDENTIFICATION; MOTILITY;
D O I
10.1016/j.theriogenology.2014.10.015
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 <= 1 x 10(-3)), 12 were upregulated in frozen-thawed sperm, the remaining 3 were upregulated in fresh sperm, and 4 other clones were identified as unknown because of incomplete sequences or because there was no significant 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.
引用
收藏
页码:504 / 511
页数:8
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