Riding the Spermatogenic Wave: Profiling Gene Expression Within Neonatal Germ and Sertoli Cells During a Synchronized Initial Wave of Spermatogenesis in Mice
Riding the Spermatogenic Wave: Profiling Gene Expression Within Neonatal Germ and Sertoli Cells During a Synchronized Initial Wave of Spermatogenesis in Mice
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DOI：
10.1095/biolreprod.114.118034
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发表时间：
2014-05-01
影响因子：
3.6
通讯作者：
Griswold, Michael
中科院分区：
文献类型：
--
作者：
Evans, Elizabeth;Hogarth, Cathryn;Griswold, Michael
精子的持续产生依赖于异步进行精子发生的生殖细胞。因此，睾丸始终含有处于分化过程不同阶段的混合生殖细胞群。睾丸的异质性使得在评估野生型睾丸时不可能分析支持细胞或特定生殖细胞群体内的基因表达。我们最近报道了一种独特的方法，通过操纵新生儿睾丸内的 RA 水平来同步精子发生而不影响生育能力。使用该方案，结合 RiboTag 转基因小鼠系，我们在精子发生的初始同步波期间绘制了支持细胞和生殖细胞翻译组图。通过微阵列分析，我们分别鉴定了 392 个和 194 个生殖细胞和支持细胞转录本，它们在精原细胞分化、分裂和减数分裂开始期间动态变化。功能注释聚类显示，生殖细胞中富集的转录本主要与减数分裂（21 个转录本）、染色质组织（12 个转录本）和细胞周期（3 个转录本）相关。此外，糖蛋白（65 个转录本）、细胞粘附（15 个转录本）和细胞连接（13 个转录本）转录本在支持细胞富集列表中过多。这些数据集代表了精原细胞分化、分裂和减数分裂开始的首次转录分析。这些数据表明，在生殖细胞进入减数分裂之前，一些编码减数分裂蛋白的基因就已表达并正在积极翻译。此外，这项研究还提供了新的候选基因 Asf1b 和 Esyt3，它们可能参与精原细胞染色质重组、生殖支持细胞相互作用和/或血睾屏障形成的调节。
Continual sperm production relies on germ cells undergoing spermatogenesis asynchronously. As a result, the testis always contains a mixed population of germ cells at different stages of their differentiation process. The heterogeneous nature of the testis makes profiling gene expression within Sertoli cells or specific populations of germ cells impossible when a wild-type testis is assessed. We recently reported a unique method for synchronizing spermatogenesis without affecting fertility by manipulating RA levels within the neonatal testis. Using this protocol, combined with the RiboTag transgenic mouse line, we have mapped the Sertoli and germ cell translatome during the initial synchronized wave of spermatogenesis. Using microarray analysis, we identified 392 and 194 germ cell and Sertoli cells transcripts, respectively, that dynamically change during spermatogonial differentiation, division, and the onset of meiosis. Functional annotation clustering revealed that transcripts enriched in germ cells were mostly associated with meiosis (21 transcripts), chromatin organization (12 transcripts), and cell cycle (3 transcripts). In addition, glycoproteins (65 transcripts), cell adhesion (15 transcripts), and cell junction (13 transcripts) transcripts were overrepresented in the Sertoli cell-enriched list. These datasets represent the first transcriptional analysis of spermatogonial differentiation, division, and meiotic onset. These data suggest that several of the genes encoding meiotic proteins are expressed and are actively being translated well before germ cells enter meiosis. In addition, this study provides novel candidate genes, Asf1b and Esyt3, that may be involved in the regulation of spermatogonial chromatin reorganization, germ-Sertoli cell interactions, and/or blood-testis barrier formation.