Department of Anatomy and Cell Biology, Justus-Liebig-University of Giessen, Giessen, Germany.
Syst Biol Reprod Med. 2011 Aug;57(4):190-6. doi: 10.3109/19396368.2011.569906. Epub 2011 May 9.
Non-obstructive azoospermia (NOA) is currently evaluated by the use of conventional histopathological methods. In some cases, focal spermatogenesis is present in the testes of patients with NOA which may be almost undetectable by routine histopathological examinations. Application of molecular markers in semen to predict the spermatogenesis status in the testis will emphasize the probability of finding sperm in NOA testis through further search using TESE or mTESE. Detection of germ cell-specific transcripts in semen is a signal of germ cells present in the testis. In this study, we used molecular methods to evaluate spermatogenesis status in azoospermic men. Semen samples were collected from 203 men with azoospermia. Total RNA was extracted from the semen precipitates. First-strand complementary deoxyribonucleic acid (cDNA) was synthesized by reverse transcriptase then, (RT)-PCRs were carried out using primers for testis stage-specific genes (DAZ, AKAP4, PRM1, and PRM2). Testicular tissue biopsies were used for evaluating spermatogenesis status in testis. Histopathological examination and LH, FSH, and testosterone level measurements (chemiluminescence assay) were performed. The presence of DAZ and PRM2 transcripts in semen significantly indicated the presence of spermatogonia and spermatids in the testicular tissues. Absence of all four markers in semen confirmed the histopathological results corresponding to sertoli cell only syndrome (SCO). Although TESE should not be excluded solely on this criteria, using PRM1, PRM2, AKAP4, and DAZ transcripts in semen would provide a non-invasive molecular diagnostic tool to better counsel patients before undergoing TESE.
非阻塞性无精子症（NOA）目前通过常规组织病理学方法进行评估。在某些情况下，NOA 患者的睾丸中存在局灶性精子发生，这可能通过常规组织病理学检查几乎无法检测到。在精液中应用分子标记物来预测睾丸中的精子发生状态，将通过进一步使用 TESE 或 mTESE 进行搜索来强调在 NOA 睾丸中找到精子的可能性。检测精液中生殖细胞特异性转录本是睾丸中存在生殖细胞的信号。在这项研究中，我们使用分子方法评估无精子症男性的精子发生状态。从 203 名无精子症男性中收集精液样本。从精液沉淀物中提取总 RNA。用逆转录酶合成第一链互补脱氧核糖核酸（cDNA），然后使用睾丸阶段特异性基因（DAZ、AKAP4、PRM1 和 PRM2）的引物进行 RT-PCR。对睾丸组织活检进行评估以确定睾丸中的精子发生状态。进行组织病理学检查以及 LH、FSH 和睾酮水平测量（化学发光测定法）。精液中 DAZ 和 PRM2 转录本的存在显著表明睾丸组织中存在精原细胞和精子细胞。精液中四种标志物均不存在证实了与唯支持细胞综合征（SCO）相对应的组织病理学结果。尽管不能仅凭这一标准排除 TESE，但在精液中使用 PRM1、PRM2、AKAP4 和 DAZ 转录本将提供一种非侵入性的分子诊断工具，以便在进行 TESE 之前更好地为患者提供咨询。
