Johnston Daniel S, Wright William W, Dicandeloro Paul, Wilson Ewa, Kopf Gregory S, Jelinsky Scott A
Contraception, Women's Health and Musculoskeletal Biology, Wyeth Research, 500 Arcola Road, Collegeville, PA 19426, USA.
Proc Natl Acad Sci U S A. 2008 Jun 17;105(24):8315-20. doi: 10.1073/pnas.0709854105. Epub 2008 Jun 10.
Mammalian spermatogenesis is a complex biological process that occurs within a highly organized tissue, the seminiferous epithelium. The coordinated maturation of spermatogonia, spermatocytes, and spermatids suggests the existence of precise programs of gene expression in these cells and in their neighboring somatic Sertoli cells. The objective of this study was to identify the genes that execute these programs. Rat seminiferous tubules at stages I, II-III, IV-V, VI, VIIa,b, VIIc,d, VIII, IX-XI, XII, and XIII-XIV of the cycle were isolated by microdissection, whereas Sertoli cells, spermatogonia plus early spermatocytes, pachytene spermatocytes, and round spermatids were purified from enzymatically dispersed testes. Microarray analysis by using Rat Genome 230 2.0 arrays identified 16,971 probe sets that recognized testicular transcripts, and 398 of these were identified as testis-specific. Expression of 1,286 probe sets were found to differ at least 4-fold between two cell types and also across the stages of the cycle. Pathway and annotated cluster analyses of those probe sets predicted that entire biological pathways and processes are regulated cyclically in specific cells. Important among these are the cell cycle, DNA repair, and embryonic neuron development. Taken together, these data indicate that stage-regulated gene expression is a widespread and fundamental characteristic of spermatogenic cells and Sertoli cells.
哺乳动物的精子发生是一个复杂的生物学过程,发生在高度组织化的生精上皮组织内。精原细胞、精母细胞和精子细胞的协调成熟表明,这些细胞及其相邻的体细胞支持细胞中存在精确的基因表达程序。本研究的目的是鉴定执行这些程序的基因。通过显微切割分离出处于周期I、II-III、IV-V、VI、VIIa,b、VIIc,d、VIII、IX-XI、XII和XIII-XIV阶段的大鼠生精小管,而从酶解分散的睾丸中纯化出支持细胞、精原细胞加早期精母细胞、粗线期精母细胞和圆形精子细胞。使用大鼠基因组230 2.0阵列进行的微阵列分析鉴定出16,971个识别睾丸转录本的探针集,其中398个被鉴定为睾丸特异性。发现1,286个探针集的表达在两种细胞类型之间以及整个周期阶段至少相差4倍。对这些探针集的通路和注释聚类分析预测,整个生物学通路和过程在特定细胞中受到周期性调节。其中重要的有细胞周期、DNA修复和胚胎神经元发育。综上所述,这些数据表明阶段调控的基因表达是生精细胞和支持细胞广泛且基本的特征。