Jelinsky Scott A, Turner Terry T, Bang Hyun J, Finger Joshua N, Solarz Mark K, Wilson Ewa, Brown Eugene L, Kopf Gregory S, Johnston Daniel S
Biological Technologies, Molecular Profiling and Biomarker Discovery, Wyeth Research, Cambridge, Massachusetts 02140, USA.
Biol Reprod. 2007 Apr;76(4):561-70. doi: 10.1095/biolreprod.106.057323. Epub 2006 Dec 13.
Regional differences along the epididymis are essential for the establishment of the luminal environment required for sperm maturation. In the current study, 19 morphologically distinct segments of the rat epididymis were identified by microdissection. Total RNA was isolated from each segment and subjected to microarray analysis. Segmental analysis of epididymal gene expression identified more than 16,000 expressed qualifiers, whereas profiling of RNA from whole rat epididymis identified approximately 12,000 expressed qualifiers. Screening a panel of normal rat tissues identified both epididymal-selective and epididymal-specific transcripts. In addition, more than 3500 qualifiers were shown to be present and differentially upregulated or downregulated by more than fourfold between any two segments. The present study complements our previous segment-dependent analysis of gene expression in the mouse epididymis and allows for comparative analyses between datasets. A total of 492 genes was shown to be present on both the MOE430 (mouse) and RAE230_2 (rat) microarrays, expressed in the epididymis of both species, and differentially expressed by more than fourfold in between segments in each species. Moreover, in-depth quantitative RT-PCR analysis of 36 members of the beta defensin gene family showed highly conserved patterns of expression along the lengths of the mouse and rat epididymides. These analyses elucidate global gene expression patterns along the length of the rat epididymis and provide a novel evaluation of conserved and nonconserved gene expression patterns in the epididymides of the two species. Furthermore, these data provide a powerful resource for the research community for future studies of biological factors that mediate sperm maturation and storage.
附睾各区域的差异对于建立精子成熟所需的管腔环境至关重要。在本研究中,通过显微切割鉴定出大鼠附睾19个形态学上不同的节段。从每个节段分离总RNA并进行微阵列分析。附睾基因表达的节段分析鉴定出超过16,000个表达特征,而来自整个大鼠附睾的RNA分析鉴定出约12,000个表达特征。对一组正常大鼠组织进行筛选,鉴定出附睾选择性和附睾特异性转录本。此外,超过3500个特征在任意两个节段之间呈现差异上调或下调四倍以上。本研究补充了我们之前对小鼠附睾基因表达的节段依赖性分析,并允许对数据集进行比较分析。共有492个基因在MOE430(小鼠)和RAE230_2(大鼠)微阵列上均有显示,在两个物种的附睾中均有表达,且在每个物种的节段之间差异表达超过四倍。此外,对β-防御素基因家族36个成员的深入定量RT-PCR分析显示,小鼠和大鼠附睾全长的表达模式高度保守。这些分析阐明了大鼠附睾全长的整体基因表达模式,并对两个物种附睾中保守和非保守的基因表达模式进行了新的评估。此外,这些数据为研究界提供了强大的资源,用于未来对介导精子成熟和储存的生物学因素的研究。