Human Genetics Research Group, IDIBAPS, University of Barcelona, Hospital Clínic, C/Casanova 143, Barcelona 08036, Spain.
Hum Reprod. 2012 May;27(5):1431-8. doi: 10.1093/humrep/des021. Epub 2012 Feb 21.
Alterations in RNAs present in sperm have been identified using microarrays in teratozoospermic patients and in other types of infertile patients. However, so far, there have been no reports on using microarrays to determine the RNA content of sperm from asthenozoospermic patients.
We started the present project with the goal of characterizing the RNA abundance in the sperm cells of asthenozoospermic patients when compared with controls. To reach this objective, we initially selected four normal fertile donors and four asthenozoospermic infertile patients. Equal amounts of RNA were extracted from the sperm samples, subjected to different quality controls and hybridized to the Affymetrix U133 Plus version 2 arrays.
Several transcripts were identified that were present in different abundance in patients compared with controls. Subsequently, we validated the differential expression of three of the detected transcripts (ANXA2, BRD2 and OAZ3), using real-time PCR in a larger set of samples. A positive correlation between the expression of these transcripts and progressive motility was observed.
The sperm cells of asthenozoospermic patients contain an altered amount of some RNAs as detected using microarray analysis and subsequently validated using real-time PCR. These results open up the possibility to investigate the implication of these genes in the pathogenic mechanisms in asthenozoospermia and to consider their potential utility as infertility biomarkers.
使用微阵列技术已经在畸形精子症患者和其他类型的不育患者中鉴定了精子中存在的 RNA 变化。然而，到目前为止，还没有关于使用微阵列来确定弱精子症患者精子中 RNA 含量的报道。
我们开始了本项目，旨在比较正常生育能力的供体和弱精子症不育患者的精子细胞中的 RNA 丰度。为了实现这一目标，我们最初选择了 4 名正常生育能力的供体和 4 名弱精子症不育患者。从精子样本中提取等量的 RNA，进行不同的质量控制，并与 Affymetrix U133 Plus 版本 2 阵列杂交。
与对照组相比，在患者中发现了几种丰度不同的转录本。随后，我们使用实时 PCR 在更大的样本集中验证了检测到的三个转录本（ANXA2、BRD2 和 OAZ3）的差异表达。这些转录本的表达与前向运动呈正相关。
使用微阵列分析检测到，弱精子症患者的精子细胞中含有一些 RNA 的含量发生了改变，随后使用实时 PCR 进行了验证。这些结果为研究这些基因在弱精子症发病机制中的作用以及考虑它们作为不育症生物标志物的潜在用途提供了可能性。
