title: Comparative transcript profiling of gene expression of fresh and frozen-thawed bull sperm.
abstract: 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 (|score (d)| ≥ 2, fold change > 1, and false discovery rate < 0.05). Of 15 differentially expressed unigenes exhibited high sequence homology (E-value ≤ 1 × 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.
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilization of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Chen
authors.affiliation: Jilin Agriculture University, Changchun 130118, China.
authors.forename: Yonggui
authors.initials: Y
authors.lastname: Wang
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilization of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Huabin
authors.initials: H
authors.lastname: Zhu
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilization of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Haisheng
authors.initials: H
authors.lastname: Hao
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilization of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Xueming
authors.initials: X
authors.lastname: Zhao
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilization of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Tong
authors.initials: T
authors.lastname: Qin
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilization of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China. Electronic address: dwangcn2002@vip.sina.com.cn.
authors.forename: Dong
authors.initials: D
authors.lastname: Wang
mesh_terms: D000818:Animals; D002417:Cattle / Q000502:physiology*; D015925:Cryopreservation / Q000662:veterinary; D005615:Freezing; D005786:Gene Expression Regulation / Q000502:physiology; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000235:genetics / Q000378:metabolism; D012662:Semen Preservation / Q000662:veterinary*; D013094:Spermatozoa / Q000378:metabolism*; D059467:Transcriptome
chemical_list: D012313:RNA
keywords: Bull; Differentially expressed genes; Fresh and frozen-thawed sperm; SSH; cDNA microarray