Screening for biomarkers of spermatogonia within the human testis: a whole genome approach
Screening for biomarkers of spermatogonia within the human testis: a whole genome approach
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DOI：
10.1093/humrep/deq053
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发表时间：
2010-05-01
影响因子：
6.1
通讯作者：
Spiess, Andrej-Nikolai
中科院分区：
文献类型：
--
作者：
von Kopylow, Kathrein;Kirchhoff, Christiane;Spiess, Andrej-Nikolai
研究精原细胞生物学的关键步骤是确定其整体基因表达谱。然而，这些细胞从睾丸分离可能会在相当大的程度上改变它们的轮廓。为了研究人类精原细胞（包括精原干细胞）在其同源微环境中的分子表型，本研究利用了一种罕见的人类精子发生缺陷亚型（其中精原细胞是唯一的生殖细胞类型），在Affyellow微阵列平台上评估了这些样本的整体表达谱，并与显示单一Sertoli细胞外观的组织进行了比较;通过实时定量PCR和免疫组化对不同的样本集进行了筛选，精原细胞的出现与基因表达水平的高度显著性差异相关，包括239个最佳候选人的人类精原细胞表达基因。具体地，成纤维细胞生长因子受体3（FGFR 3）、桥粒芯糖蛋白2（DSG 2）、E3泛素连接酶c-CBL（Casitas B细胞淋巴瘤），癌症/睾丸抗原NY-ESO-1显示CTAG 1A/B、未分化胚胎细胞转录因子1（UTF 1）和突触体相关蛋白91 kDa同源物（SNAP 91）代表人精原细胞的特异性生物标志物。特别是表面标志物FGFR 3和DSG 2，可能有助于人精原干细胞和/或祖细胞的分离和富集，从而为人精原干细胞/祖细胞的长期维持、精原细胞自我更新、克隆扩增和分化的研究奠定基础。
A key step in studying the biology of spermatogonia is to determine their global gene expression profile. However, disassociation of these cells from the testis may alter their profile to a considerable degree. To characterize the molecular phenotype of human spermatogonia, including spermatogonial stem cells (SSCs), within their cognate microenvironment, a rare subtype of human defective spermatogenesis was exploited in which spermatogonia were the only germ cell type.The global expression profile of these samples was assessed on the Affymetrix microarray platform and compared with tissues showing homogeneous Sertoli-cell-only appearance; selected genes were validated by quantitative real-time PCR and immunohistochemistry on disparate sample sets.Highly significant differences in gene expression levels correlated with the appearance of spermatogonia, including 239 best candidates of human spermatogonially expressed genes. Specifically, fibroblast growth factor receptor 3 (FGFR3), desmoglein 2 (DSG2), E3 ubiquitin ligase c-CBL (casitas B-cell lymphoma), cancer/testis antigen NY-ESO-1 (CTAG1A/B), undifferentiated embryonic cell transcription factor 1 (UTF1) and synaptosomal-associated protein, 91 kDa homolog (SNAP91) were shown to represent specific biomarkers of human spermatogonia.These biomarkers, specifically the surface markers FGFR3 and DSG2, may facilitate the isolation and enrichment of human stem and/or progenitor spermatogonia and thus lay a foundation for studies of long-term maintenance of human SSCs/progenitor cells, spermatogonial self-renewal, clonal expansion and differentiation.