Department of Andrology, Center for Men's Health, Department of ART, Institute of Urology, Urologic Medical Center, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, China.
School of Medicine, Hunan Normal University, 371 Tongzipo Road, Changsha, Hunan 410013, China; Shanghai Key Laboratory of Reproductive Medicine, Shanghai 200025, China.
Tissue Cell. 2019 Jun;58:24-32. doi: 10.1016/j.tice.2019.04.003. Epub 2019 Apr 6.
The proper assessment of male fertility is essential for diagnosing and treating male infertility. Currently, spermiogram and Johnsen testicular biopsy score counts are used to assess male fertility. However, spermiogram is not a suitable option for non-obstructive azoospermia patients, and Johnsen testicular biopsy scores only represent localized and not the overall spermatogenesis. Whole-mount staining was a novel method for evaluating protein expression in the tissue. Thus, we explored its application in human seminiferous tubules. Testicular biopsies from 57 azoospermia patients were categorized as obstructive azoospermia (OA), maturation arrest (MA) and Sertoli-cells only syndrome (SCOS). We performed whole-mount staining of their seminiferous tubules and evaluated the spermatogonial stem cells (SSCs), differentiated spermatogonia (SG), spermatocytes (SPC) and spermatids (SD) with their respective markers (GFRA1, CD117, SYCP3, and PNA) to assess fertility. GFRA1, CD117, SYCP3, and PNA were not expressed in SCOS patients, whereas all of them were detected in OA patients. In MA patients with arrested spermatogenesis at the SPC stage, GFRA1, CD117, and SYCP3, but not PNA were expressed in the seminiferous tubules. In MA patients with arrested spermatogenesis at the spermatogonia stage, only GFRA1 was expressed in the seminiferous tubules. These results were consistent with the Johnsen testicular biopsy score counts except for one patient, where although only Sertoli cells were indicated by the score, SSCs were also detected in the whole-mounts. Collectively, whole-mount staining could be used to analyze the inherent spermatogenesis of seminiferous tubules through staining of germ cells at different stages. It offers a more accurate and promising faster method for assessing male fertility compared with traditional biopsy screening. And it could have potential value for the clinical purpose for male fertility management.
男性生育力的正确评估对于男性不育的诊断和治疗至关重要。目前，精子分析和约翰森睾丸活检评分用于评估男性生育力。然而，精子分析并不适用于非梗阻性无精子症患者，而约翰森睾丸活检评分仅代表局部而非整体生精功能。全组织染色是一种评估组织中蛋白质表达的新方法。因此，我们探索了其在人类曲细精管中的应用。对 57 例无精子症患者的睾丸活检进行分类，分为梗阻性无精子症（OA）、成熟阻滞（MA）和唯支持细胞综合征（SCOS）。我们对其曲细精管进行全组织染色，并使用各自的标记物（GFRA1、CD117、SYCP3 和 PNA）评估精原干细胞（SSCs）、分化精原细胞（SG）、精母细胞（SPC）和精子（SD），以评估生育能力。GFRA1、CD117、SYCP3 和 PNA 在 SCOS 患者中不表达，而在 OA 患者中均有表达。在精母细胞阻滞于 SPC 期的 MA 患者中，GFRA1、CD117 和 SYCP3，但无 PNA 在曲细精管中表达。在精原细胞阻滞于精原细胞期的 MA 患者中，只有 GFRA1 在曲细精管中表达。这些结果与约翰森睾丸活检评分计数一致，但有一位患者除外，尽管评分仅显示支持细胞，但全组织染色中也检测到 SSCs。总之，全组织染色可以通过对不同阶段的生殖细胞进行染色来分析曲细精管的固有生精功能。与传统的活检筛查相比，它为评估男性生育力提供了一种更准确、更有前途的快速方法。并且它可能具有男性生育力管理的临床应用价值。
