评估精子受孕能力的外显子标记物筛选

doi:10.3969/j.issn.1000-8535.2021.05.011

广州医药 ›› 2021, Vol. 52 ›› Issue (5): 52-56.DOI: 10.3969/j.issn.1000-8535.2021.05.011

评估精子受孕能力的外显子标记物筛选

吴瑞珊, 张梦媛, 刘晓华, 赵文忠

广东省计划生育科学技术研究所(广东省生殖医院)国家卫生健康委员会男性生殖与遗传重点实验室(广州 510600)

收稿日期:2021-03-09 出版日期:2021-09-20 发布日期:2021-11-24
通讯作者: 赵文忠,E-mail:zhaowzphd@163.com
基金资助:
广州市科技计划项目(201804010431)

Screening of exon marker to evaluate the fertilizing ability of sperm

WU Ruishan, ZHANG Mengyuan, LIU Xiaohua, ZHAO Wenzhong

NHC Key Laboratory of Male Reproduction and Genetics, Guangdong Provincial Reproductive Science Institte (Guangdong Provincial Fertility Hospital),Guangzhou 510600,China

Received:2021-03-09 Online:2021-09-20 Published:2021-11-24

摘要/Abstract

摘要： 目的通过对不同受孕能力精子外显子的分析,寻找并验证特异性外显子作为精子受孕能力的生物标记物。方法基于二代测序数据进行生物信息学分析,寻找特异性外显子并设计引物。各取8份高、低受孕能力精液标本,提取精子RNA反转录后进行real time q-PCR验证外显子的表达效率,筛选表达差异恒定的精子外显子作为生物标记物。各取10份高、低受孕能力精子标本,用筛选后的外显子引物进行real time q-PCR验证。结果生物信息学分析得到31个候选精子外显子,从31个候选外显子中筛选出9个表达差异恒定的精子外显子GAPDHS、HSF2BP、HSPA1L、ADAM21、SPEM1、WBP2NL、DDX20、TSGA10、PGK2;real time q-PCR验证结果显示,在高、低受孕能力精液标本中这9种精子外显子表达差异明显。结论初步确定,差异表达恒定的九种外显子可作为评估精子质量的生物标记物。

关键词: 外显子, 精子, 受孕能力

Abstract: Objective To find and verify specific exons as biomarkers of sperm fertility by analyzing sperm exons with different fertility ability.Methods Based on the second generation sequencing data, bioinformatics analysis was conducted to find specific exons and design primers. We obtained 16 semen samples, 8 of high and the other 8 of low fertilizing ability, after the sperm RNAs were extracted and reverse-transcribed, real time q-PCR was performed to verify the expression efficiency of exons, and the sperm exons with constant expression difference were selected as biomarkers. 10 high and 10 lowfertility ability sperm samples were taken for real time q-PCR verification with screened exon primers. Results Thirty-one sperm exons were obtained by bioinformatics analysis, and 9 sperm exons with constant expression differences were selected from the 31 candidate exons, including GAPDHS, HSF2BP, HSPA1L, ADAM21, SPEM1, WBP2NL, DDX20, TSGA10 and PGK2. The results of real time q-PCR verification showed that the exons of these 9 sperm were significantly different in the semen samples with high and low fertility ability. Conclusion Nine exons with constant differential expression can be used as biomarkers to evaluate sperm quality.

Key words: exon, sperm, fertilizing ability

吴瑞珊, 张梦媛, 刘晓华, 赵文忠. 评估精子受孕能力的外显子标记物筛选[J]. 广州医药, 2021, 52(5): 52-56.

WU Ruishan, ZHANG Mengyuan, LIU Xiaohua, ZHAO Wenzhong. Screening of exon marker to evaluate the fertilizing ability of sperm[J]. Guangzhou Medical Journal, 2021, 52(5): 52-56.

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评估精子受孕能力的外显子标记物筛选

Screening of exon marker to evaluate the fertilizing ability of sperm

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