Zhu Zijue, Li Chong, Yang Shi, Tian Ruhui, Wang Junlong, Yuan Qingqing, Dong Hui, He Zuping, Wang Shengyue, Li Zheng
Department of Urology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 1630 Dongfang Road, Shanghai 200127, China.
Department of Andrology, Urologic Medical Center, Shanghai General Hospital, Shanghai Jiao Tong University, 100 Haining Road, Shanghai 200080, China.
Sci Rep. 2016 Jan 12;6:19069. doi: 10.1038/srep19069.
Many infertile men are the victims of spermatogenesis disorder. However, conventional clinical test could not provide efficient information on the causes of spermatogenesis disorder and guide the doctor how to treat it. More effective diagnosis and treating methods could be developed if the key genes that regulate spermatogenesis were determined. Many works have been done on animal models, while there are few works on human beings due to the limited sample resources. In current work, testis tissues were obtained from 27 patients with obstructive azoospermia via surgery. The combination of Fluorescence Activated Cell Sorting and Magnetic Activated Cell Sorting was chosen as the efficient method to sort typical germ cells during spermatogenesis. RNA Sequencing was carried out to screen the change of transcriptomic profile of the germ cells during spermatogenesis. Differential expressed genes were clustered according to their expression patterns. Gene Ontology annotation, pathway analysis, and Gene Set Enrichment Analysis were carried out on genes with specific expression patterns and the potential key genes such as HOXs, JUN, SP1, and TCF3 which were involved in the regulation of spermatogenesis, with the potential value serve as molecular tools for clinical purpose, were predicted.
许多不育男性是精子发生障碍的受害者。然而,传统的临床检测无法提供关于精子发生障碍病因的有效信息,也无法指导医生如何进行治疗。如果能确定调控精子发生的关键基因,就可以开发出更有效的诊断和治疗方法。在动物模型方面已经开展了许多工作,但由于样本资源有限,在人类方面的研究较少。在当前的研究中,通过手术从27例梗阻性无精子症患者获取睾丸组织。选择荧光激活细胞分选和磁激活细胞分选相结合的方法,作为在精子发生过程中对典型生殖细胞进行分选的有效方法。进行RNA测序以筛选精子发生过程中生殖细胞转录组图谱的变化。差异表达基因根据其表达模式进行聚类。对具有特定表达模式的基因以及参与精子发生调控的潜在关键基因如HOXs、JUN、SP1和TCF3进行基因本体注释、通路分析和基因集富集分析,并预测其作为临床分子工具的潜在价值。